DNA REPAIR CATALOG OF ANTIBODIES FOR. FAX: WEB:

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1 CATALOG OF ANTIBODIES FOR DNA REPAIR For research purposes only. Not for use in humans. Prices subject to change. FAX: WEB:

2 Table of Contents Non-Homologous End Joining...2 Homologous Recombination How To Series...3 Checkpoint Signaling & Double Strand Break Repair Base Excision Repair...8 Nucleotide Excision...9 Mismatch Repair Antibody Grants DNA Polymerase Direct Reversal Longevity Syndromes Linked to DNA Repair Defects In the News Application Key ELISA - Elisa FACS - Fluorescent Activated Cell Sorting DNA Repair DNA repair refers to a collection of processes by which a cell identifies and corrects damage to the DNA molecules that encode its genome. In human cells, both normal metabolic activities and environmental factors such as UV light can cause DNA damage, resulting in up to half a million individual molecular lesions per cell per day. Many of these lesions cause structural damage to the DNA molecule and can alter or eliminate the cell s ability to transcribe the gene that the affected DNA encodes. Other lesions induce potentially harmful mutations in the cell s genome, which affect the survival of its daughter cells after it undergoes mitosis. Consequently, the DNA repair process must be constantly active so it can respond rapidly to any damage in the DNA structure. The rate of DNA repair is dependent on many factors, including the cell type, the age of the cell, and the extracellular environment. A cell that has accumulated a large amount of DNA damage, or one that no longer effectively repairs damage incurred to its DNA, can enter one of three possible states: 1. an irreversible state of dormancy, known as senescence 2. cell suicide, also known as apoptosis or programmed cell death 3. unregulated cell division, which can lead to the formation of a tumor that is cancerous The DNA repair ability of a cell is vital to the integrity of its genome and thus to its normal functioning and that of the organism. Many genes that were initially shown to influence lifespan have turned out to be involved in DNA damage repair and protection. Failure to correct molecular lesions in cells that form gametes can introduce mutations into the genomes of the offspring and thus influence the rate of evolution. ICC - Immunocytochemistry IF - Immunofluorescence IHC - Immunohistochemistry exogenous damage endogenous Metabolism damage healthy cell rate of DNA damage = rate of repair IHC-Fr - Immunohistochemistry Frozen IHC-P - Immunohistochemistry Paraffin IP - Immunoprecipitation - Western Blot Reactivity Key Bv - Bovine Mk - Monkey Ca - Canine Mu - Ce - C. Elegans Rb - Ch - Chicken Rt - Rat Dr - Drosophila Ye - Yeast Ft - Ferret Vi - Virus Ha - Hamster Xp - Xenopus - man cancer repair nuclear DNA unrepaired senesence Damage Pathology repair mitochondrial DNA unrepaired apoptosis up to 500,000 DNA modification events per cell per day diseased cell rate of DNA damage > rate of repair 1

3 Non-Homologous End Joining (NHEJ) Antibodies Non-homologous end joining (NHEJ) is a pathway that can be used to repair double-strand breaks in DNA. NHEJ is referred to as non-homologous because the break ends are directly ligated without the need for a homologous template, in contrast to homologous recombination, which requires a homologous sequence to guide repair. NHEJ is evolutionarily conserved throughout all kingdoms of life and is the predominant double-strand break repair pathway in many organisms, including higher eukaryotes such as human and mouse. NHEJ typically utilizes short homologous DNA sequences, termed microhomologies, to guide repair. Microhomologies in the single-stranded overhangs that are often present on the ends of double-strand breaks are used to promote restorative repair. When these overhangs are compatible, NHEJ almost always repairs the break accurately, with no sequence loss. Imprecise repair leading to loss of nucleotides can also occur, but is much less common. Nevertheless, NHEJ is often somewhat misleadingly referred to as an error-prone repair mechanism. This is likely because NHEJ can lead to translocations when organisms are subjected to large doses of radiation that cause many breaks per cell. Additionally, the NHEJ pathway is responsible for fusing the ends of chromosomes that have undergone telomere failure. These translocations may result in incorrect gene regulation, leading to pathological conditions. NB Artemis NB Artemis Chicken IHC, NB Artemis NB Ku70 NB Ku70 (N3H10) IHC, IF, IP, Bv(-), Ha,, Mu, Mk, Rb(-), Xp NB Ku70 NB Ku70 (1.5) NB Ku70 IHC-Fr, IHC-P, IP,, Mu, Rt, Ha, Xp NB Ku80 (5D5.8) NB Ku80 (9403) NB Ku80, Mu, Ha, Rt NB Ku80, Ha, Mu, Rt NB Ku80 Goat ELISA, NB LIG4 ELISA,, IHC-P H M02 PRKDC (1B9), ELISA NB PRKDC/DNA-PK (Y393), ICC, IHC, Mu(-), Rt(-) NB XRCC4 NB XRCC4 IP,, Rt NB XRCC4 IF, SAMPLE SIZES AVAILABLE Ku70/Ku80 DNAPKcs XRCC4-LIG4-XLF Nuclease Artemis NB of Artemis protein in human testis lysate (NB ). Ku70 NB of human Ku70 in HeLa lysates LIG4 NB Staining of human pancreatic tissue. XRCC4 NB of XRCC4 in HeLa nuclear extracts (NB800-PC9). Pathway Description: Ku70/80 binds to DNA ends and recruits other NHEJ proteins to the break. DNAPKcs activates nucleases via phosphorylation to repair any damaged DNA. Finally the repaired DNA is ligated by the XRCC4/Ligase IV complex to complete the re-closure of the DNA break. 2

4 Homologous Recombination Antibodies Double-Strand Breaks are perhaps the most serious form of DNA damage because they pose problems for transcription, replication, and chromosome segregation. DSBs differ from most other types of DNA lesions in that they affect both strands of the DNA duplex and therefore prevent use of the complementary strand as a template for repair. Failure to repair these defects can result in chromosomal instabilities leading to deregulated gene expression and carcinogenesis. To counteract the detrimental effects of these potent lesions, cells have evolved two distinct pathways of DSB repair, Homologous Recombination (HR) and NonHomologous End Joining (NHEJ). In the process of HR, a DSB is repaired by copying the missing information from the sister chromatid or homologous chromosome, resulting in the exact restoration of the DNA. (Ref.1). Catalog# NB Product BRCA1 Host Type Application, IP Species NB BRCA1 (6B4), IP NB BRCA1 [Ser1189] NB BRCA1 [Ser1280] NB BRCA1 [Ser1387] NB BRCA1 [Ser1423], FACS, Mu NB BRCA1 [Ser1457] NB BRCA1 [Ser1466] NB BRCA1 [Ser1524] NB DMC1 (2H12/4) IF, IP,, Mu, Rt, Bv NB DMC1 (5B10/2), IP NB EME1 (mta317 h2/1) NB Mre11, IP, IF, IHC-P, Mu NB Mre11 NB Mre11 (12D7), IP, IF, IHC-P NB NBS1, IP, IF, Mu(-) NB NBS1, IP NB NBS1, IP NB NBS1, IHC, Mu, Rt NB NBS1 (1C3), IP NB NB NBS1 [Ser343], IF, IHC Mu NB NBS1 (Y1121), IP, IF, IHC-P, Mu, Rt(-) NB RAD50, IF(-), IP, Mu NB RAD50, IP NB RAD50 IHC, Mu 1. Jackson SP. Sensing and repairing DNA doublestrand breaks. Carcinogenesis May; 23(5): Review. BRCA1 NB Mre11 NB IP/Western blot of human BRCA1 in HeLa lysates Staining of the human epidermis. NBS1 NB NBS1 NB of human NBS1 in HeLa lysates of human NBS1 in HeLa (NB800PC1) and HepG2 lysates. How To Series CD s Do you need to freshen up your IHC, RNAi Western Blot or ELISA skills? A little fuzzy on the theory? Have students to teach? Check out our How To Series: Go to and click on Downloads to download our four presentations. 3

5 Homologous Recombination Antibodies RAD50 NB RAD50 NB of Rad50 in Jurkat cell lysate (NB800-PC2). of Rad50 in HeLa lysate RAD51 NB RAD52 NB of RAD52 in T24 bladder carcinoma. SIRT4 NB of SIRT4 in human kidney lysate (NB ). XRCC2 NB Staining of human testis. XRCC4 NB IP/ of man Rad52 in HeLa lysates XRCC1 NB of XRCC1 in HeLa Nuclear Cell Extract (NB800-PC9). XRCC4 NB of XRCC4 in HeLa nuclear extract (NB800-PC9). Catalog# Product Host Type Application Species NB RAD50 (13B3), IF NB RAD50 (zinc hook domain), IP NB RAD51 (13E4) NB RAD51 (14B4), IP, IF H M01 RAD51 (2E5-E5), ELISA NB RAD51B (1 H3/13), Mu(-) NB RAD51B (1E11/6) Ha, NB RAD51C Goat NB RAD51C (2H11/6), IF (-), Mu H M01 RAD51C (3F3-5C6), ELISA NB RAD51D, Mu(-) NB RAD51D Goat, ELISA NB RAD51D (5B3/6), IF, Mu(-) NB RAD51L1 Goat, ELISA H M01 RAD51L3 (1C8-3C11), ELISA NB RAD52 (-), IP NB RAD52 IP NB RAD54 (4E3/1) H M01 RAD54B (4A7), ELISA H M01 RAD54L (4G2), ELISA NB SIRT4 Goat, ELISA NB XRCC1, IP, ICC NB XRCC1 (33-2-5), IHC, IF, IP, ICC, Rt NB XRCC1, IP, IHC, ELISA NB XRCC1 (Agarose) IP NB XRCC1 [Ser461] NB XRCC1 [Ser475] NB XRCC1 [S518/T519/T523] NB XRCC2 IHC-Fr, IHC-P, NB XRCC3 NB XRCC3 IHC, IP,, Mu, Rt NB XRCC3 (10F1/6), IF, Mu(-) NB XRCC4 NB XRCC4 IP,, Rt NB XRCC4 IF, NB XRCC4, IP, IHC, Mu, Rt Can t Decide? Try a SuperNovus Pack: NB Mre11-Rad50-NBS1 (3 different antibodies) SAMPLE SIZES AVAILABLE Abnova, Acris, biosensis, Innova Novus distributes for these companies: Staining of man HCT116 cells. 4

6 s1 Checkpoint Signaling & Double Strand Break Repair Antibodies The genomes of mammalian cells are under continuous assault by environmental agents (e.g., UV light and reactive chemicals) as well as the byproducts of normal intracellular metabolism (e.g., reactive oxygen intermediates and inaccurately replicated DNA). Whatever the origin, genetic damage threatens cell survival and leads to organ failure, immunodeficiency, cancer, and other pathologies sequelae. To ensure that cells pass accurate copies of their genomes on to the next generation, evolution has overlaid the core cell-cycle machinery with a series of surveillance pathways termed cell-cycle checkpoints. The overall function of these checkpoints is to detect damaged or abnormally structured DNA, and to coordinate cell-cycle progression with DNA repair. Typically, cell-cycle checkpoint activation slows or arrests cell-cycle progression, thereby allowing time for appropriate repair mechanisms to correct genetic lesions before they are passed on to the next generation of daughter cells. (1) NB BP1 IF,, ICC, IHC-P, Mu NB BP1 IF,, ICC, Mu NB BP1, IHC, Mu NB BP1 [Ser25], IP, Mu NB ATM IHC, IP, NB ATM Goat, IP, Mu NB ATM Goat, IP, Mu NB ATM Ha,, Mu, Mk NB ATM, IHC Ha,, Mu, Mk NB ATM Chicken ICC, IHC,, Mu NB ATM, IHC NB ATM IHC NB ATM (10H11.E12)[Ser1981] ELISA, IHC(-), IF,, Mu NB ATM (2C1), IP, IHC, ICC, Mu, Mk, Rt NB ATM (3E8) IP,, Mu, Mk, Rt NB ATM (5C2) IF,, Mu, Mk, Rt NB ATM (7C10D8) [Ser1981] ELISA, IHC, IF,, Mu NB / 399 ATM (Agarose) Goat IP, Mu NB ATM (ATX08) IHC, Mu NB ATM (Y170) [Ser1981] IHC, IP,, ICC, Mu(-), Rt(-) NB ATM [Ser1981], IF NB ATR IF, IP, NB ATR, Mu NB ATR, Mu NB ATR, Mu, Ha, Bv NB ATR (2B5) IF, IP, NB ATRIP, IP NB ATRIP, IP NB ATRX, IP NB ATRX IHC-P H M01 ATRX (3C9) ELISA, NB Aurora A IHC, IP, ICC(-) NB Aurora A, IF Mu, Rt NB Aurora A Goat NB Aurora A (35C1) ELISA, IF, IP,, ICC, Mu NB Aurora A (35C1) (Biotin) IF,, Mu (1) Robert T. Abraham. Genes and Development, 16(17): , G1 S1 ATR & Checkpoint Signaling PLK1 p53 TAK1 PCNA MLH1 CHK1 ATR CDC25 B/C HDAC1 ATRIP RAD9 RAD1 Claspin WEE1 CDC2 Pathway Description: DNA damage which causes in replication fork stalling or UV induced dimerization results in cell cycle checkpoint activation. Damage sensed by the 911 complex or PCNA activates ATR. The activation of ATR blocks the action of CDC25B/C and prevents the progression of the cell cycle from G2 to M phase. ATM NB Staining of Testis, Seminiferous Tubule. AURORA A NB M1 G2 Staining of human pancreatic tumor tissue. 5

7 Checkpoint Signaling & DSB Repair Antibodies NB Aurora A [Thr288] IF,, ICC BRCA1 See SeHomologousrecombination Recombination NB CDC25A, IP NB CDC25A, IP NB CDC25A Chicken ICC, NB / 419 CDC25A (Agarose) IP NB CDC25A (DCS 121) IP, NB CDC25A (SPM233) IHC-P, Rt NB CDC25A [Ser17] NB CHK1 Goat NB CHK1 NB CHK1 Goat, IP NB CHK1, IP NB CHK1 IHC NB CHK1 IHC NB CHK1 (2G11D5), ELISA NB CHK1 (DCS-300) NB CHK1 (DCS-310) IHC-P, IP,, Mu, Rt NB CHK1 (E250) FACS, IHC,, ICC, Mu, Rt(-) NB CHK1 (EP691Y) FACS, IHC,, ICC, Mu(-), Rt(-) NB CHK2 (-), IP NB CHK2, IP NB CHK2, Mu, Rt NB CHK2 (8F12) NB CHK2 (E126) [Thr68] IP,, ICC, Mu(-), Rt(-) NB CHK2 (Y171) [Thr68] IP,, Mu(-), Rt(-) NB gamma-h2ax [Ser139], IF, Mu NB HDAC1 (-), IP NB HDAC1, IP, Mu H M06 HDAC1 (1D6) ELISA,, IHC-P H M02 HDAC1 (3E1) ELISA,, IHC-P H M11 HDAC1 (5A11) ELISA, H M14 HDAC1 (5C11) ELISA,, IHC-P NB HDAC1 (HDAC1-21) ELISA, IP,, Mu Ku 70 See Non-Homologous see non-homologous End Joining Ku 80 See Non-Homologous see non-homologous End Joining NB LIG4, IHC-P, ELISA NB MLH1, IP NB MLH1 (Agarose) IP H M02 MLH1 (M1) ELISA,, IHC-P MRE11 See SeHomologousrecombination Recombination NBS1 See SeHomologousrecombination Recombination NB p53, ELISA, IHC NB p53 (BP53-12) ELISA, IHC, IP,, ICC, Mk NB p53 (BP53-12) (FITC) FACS, ICC, IHC-Fr, IHC-P, Mu NB p53 (FITC) FACS Bv, Mu(-), Rt(-) NB p53 (PAb 240) ELISA, FACS, IP, IHC-Fr, IHC-P, Mu, Rt, Xp(-) NB p53 [Ser15], IP, IHC, Mu, Rt CDC25A NB IP/ of human CDC25a in HeLa nuclear extracts (NB800-PC9). CHK1 NB Staining of paraffinembedded human breast carcinoma. CHK2 NB Staining of paraffinembedded human colon carcinoma HDAC1 H M06 Staining of HDAC1 on formalinfixed paraffinembedded human liver. MLH1 NB Staining of human gastric cancer. CHK1 NB Staining i of human testis. CHK2 NB IP/ of man CHK2 in HeLa lysates gamma-h2ax NB Immunofluorescent staining of HEK293 cells. LIG4 NB Staining of human pancreatic tissue. MLH1 H M02 Staining of human testis. 6

8 Checkpoint Signaling & DSB Repair Antibodies NB p53 [Ser392], Mk NB p53dinp1, Mu, Rt NB p53r2, Mu, Rt NB PERP NB , NB , NB PLK1 IF, IP, H M01 PLK1 (2G12) ELISA, H M02 PLK1 (3F10) ELISA, H M04 PLK1 (4G11) ELISA, NB PP2A Goat ELISA, NB PP2A, IP(-) NB PP2A, IP, Mu NB PP2A (TQ11-1G6), IP, Mu NB PP2A (Y119) IHC, IP,, ICC, Mu, Rt NB PP2A (YE351) IHC,, Mu, Rt H M02 PRKDC (1B9) ELISA, NB PRKDC/DNA-PK (Y393) IHC,, ICC, Mu(-), Rt(-) NB PUMA, IHC, Mu NB PUMA NB PUMA NB RAD17 NB RAD17 Ye NB RAD17 Goat NB RAD17 (1C6/2) Ye H M01 RAD17 (2G12) ELISA, NB RAD17 [Ser645] NB RAD17 [Ser647] IF, ICC, Mu RAD50 See SeHomologousrecombination Recombination RAD51 See SeHomologousrecombination Recombination NB RPA 14 (11.1), IP, Mu NB RPA 14 (14.1), IP, Mu NB RPA 32 NB RPA 32 (12F3.3), IP, Mu NB RPA 34 (9H8) IHC-Fr, IHC-P, IP, IF, NB SMC1, IF, Mu, Xp NB SMC1 IHC NB SMC1 (Agarose) IP NB SMC1 [Ser957], IP NB SMC1 [Ser966], IP NB SMC1 [Ser966] IHC H M01 SMC1L1 (1B9) ELISA,, IHC-P H M01 WEE1 (5B6) ELISA, XRCC4 See SeHomologousrecombination Recombination PP2A NB of PP2A in human lymph node lysate (NB ). RAD17 [Ser645] NB of human Phospho- Rad17. ATM Kinase & Checkpoint Signaling G1 S1 p21 CDK2 p53 CDC25A NBS1 MRE11 Rad50 SMC1 PLK1 CHK2 ATM PUMA NB Staining of PUMA in K562 cells SMC1 [Ser957] NB of SMC1 [ps957] in 10 Gy irradiated HeLa Aurora A TAK1 CDC25 B/C CDC2 BRCA1 Pathway Description: ATM kinase is capable of regulating cell cycle progression in response to DNA damage at multiple points. Activation of CHK2 and CDC25A is able to prevent the G1 to S transition. The action of the MRN complex and SMC1 act to prevent progression of S phase. Prevention of mitosis by ATM signaling occurs via the action of BRCA1 and inhibition of CDC25B/C. M1 G2 Can t Decide? Try a SuperNovus Pack: NB BP1 SuperNovus Pack NB ATM SuperNovus Pack NB ATR SuperNovus Pack NB Aurora-A SuperNovus Pack NB CDC25a SuperNovus Pack SAMPLE SIZES AVAILABLE 7

9 Base Excision Repair Antibodies Base excision repair (BER) is a cellular mechanism that can repair damaged DNA during DNA replication. Repairing DNA sequence errors is necessary so that mutations are not induced during replication. Single bases in DNA can be chemically mutated, for example by deamination or alkylation, resulting in incorrect base-pairing, and consequently, mutations in the DNA. Base excision repair involves flipping the mutated base out of the DNA helix and repairing the base alone. There are two main enzymes used, DNA glycosylases and AP endonucleases. The DNA glycosylase is used to break the β-n glycosidic bond to create an AP site. AP endonuclease recognizes this site and nicks the damaged DNA on the 5 side (upstream) of the AP site creating a free 3 -OH. DNA polymerase, Pol I, extends the DNA from the free 3 -OH using its exonuclease activity to replace the nucleotide of the damaged base, as well as a few downstream, followed by sealing of the new DNA strand by DNA ligase. NB APE1, IHC, Mu, Rt NB APE1 (13B8E5C2), IHC, Mu, Rt NB APE1 Goat NB DNA Ligase 1 (10H5), IP NB DNA Ligase III NB DNA Ligase III (6G9), IP, IHC NB FEN-1 (FEN-1) NB FEN-1 (3E7) NB MBD4, Mu NB MBD4 NB MTH1, IHC NB MYH NB NEIL1 Goat, ELISA H M01 NEIL2 (1B7), ELISA NB NTH1 NB NTH1 NB Ogg1, IHC, Mk, Rt NB PARP, IHC, Mu, Rt NB PARP, ELISA, Bv, Rt, Mu NB /112 PARP (C-2-10), ICC, ELISA Ch(-), Ha,, Mk, Rt NB PCNA (PC10), IP, IHC, Ye, Dr, Rt, Mu, Ch NB PCNA, IP, IHC, Mu, Rt NB PCNA (PC10), [FITC] IHC, FACS Dr,, Mu, Rt, Ye NB PCNA (PC10), [PE] IHC, FACS, Dr, Mu, Rt, Ye NB SMUG1 Goat, ELISA NB UNG, Mu, Rt H M01 UNG (4C12) ELISA XRCC1 See see HomologousRecombinat Recombination ion APE1 NB DNA Ligase 1 NB PARP NB Base Excision Repair Short Patch Lig III XRCC1 Lig III XRCC1 APE1 hogg1 Pol β Pol β Pol β XRCC1 Lig III hogg1 APE1 APE1 Pol β Staining of APE/Ref-1 in prostate cancer. of DNA Ligase I in Raji whole cell lysate (NB800-PC11). of PARP in HL-60 whole cell extracts. (1) Normal (2) Induced Lig III XRCC1 Pol β 8

10 Nucleotide Excision Antibodies Nucleotide excision repair (NER) is an important DNA repair mechanism by which the cell repairs DNA damage occuring to bases. Base damage can be caused by a variety of sources including chemicals and ultraviolet (UV) light from the sun. NER is the mechanism by which the cell can prevent unwanted mutations by removing the majority of UV-induced DNA damage (mostly in the form of thymine dimers and 6-4-photoproducts). The importance of this repair mechanism is evidenced by the severe human diseases that result from in-born genetic mutations of NER proteins including Xeroderma pigmentosum and Cockayne s syndrome. NER recognizes bulky distortions in the shape of the DNA double helix. Recognition of these distortions leads to the removal of a short single-stranded DNA segment that includes the lesion, creating a single-strand gap in the DNA, which is subsequently filled in by DNA polymerase, using the undamaged strand as a template. ERCC1 H M01 CCNH [1B8], IHC, ELISA NB NB CDK7 [MO-1.1] H A01 CDK7, ELISA H A02 CETN2, ELISA NB DDB1 Goat, Mu, Rt NB DDB1, IP, Mu H A01 DDB1 ELISA, NB DNA Ligase 1 [10H5], IP NB ERCC1 [3H11], IP NB ERCC1 [8F1], IP, IHC, Rt H M01 ERCC2 [4G2-2A6] ELISA, H A01 ERCC3 ELISA, H A01 ERCC6 ELISA, H M01 ERCC8 [2G3-C6] ELISA H M01 GTF2H1 [1F12-1B5], IP, IHC H A01 GTF2H3 ELISA, NSB726 IKB alpha [Ser32/Ser36] H M01 LIG1 [10G12] ELISA, H M01 RAD23A [3C12] ELISA, RPA14 See Checkpoint See checkpoint Signaling NB RPA32 NB RPA32, IP NB RPA32, Mu NB RPA32 [Ser4/Ser8], IP, IF NB RPA32 (Ser33), IP, IF NB RPA34 [9H8], IP, IF, IHC NB RPA34 [SPM316] IHC NB RPA70, IP NB XAB2 NB XAB2 Goat H M01 XAB2 [1D1-1A9], IHC, ELISA NB XPA [12F5], IHC NB XPA, IHC NB XPC [3.26], IHC NB XPC IP NB XPG [8H7], IP of ERCC1 in HeLa whole cell lysate RPA32 NB Staining i of phosphorylated RPA32 to foci following treatment with CPT. XAB2 NB of XAB2 in A431 lysate (NB800-PC12). XPC NB Staining of HeLa cells. ERCC1 NB Staining of human tonsil. RPA70 NB of RPA70 in HeLa lysates XAB2 H M01 Staining of XAB2 on formalin-fixed paraffin-embedded human lung. XPC NB IP/ of man XPC in HeLa lysates 9

11 Mismatch Repair Antibodies Any mutational event that disrupts the superhelical structure of DNA carries with it the potential to compromise the genetic stability of a cell. Mismatch repair is a system for recognizing and repairing the erroneous insertion, deletion and misincorporation of bases that can arise during DNA replication and recombination, as well as repairing some forms of DNA damage. The fact that the damage detection and repair systems are as complex as the replication MLH1 NB Staining of human testis. MSH2 NB of human MSH2 in HeLa lysates MSH6 NB Staining ing of human MSH6 on HCT-116 cells. MSH2 NB Staining of human colon adenocarcinoma. MSH2 NB of MSH2 in human (HeLa) and mouse (NIH3T3) lysates. MSH6 NB of human MSH6 in TK6 and MT1 lyastes. Antibody Grants machinery itself highlights the importance evolution has attached to DNA fidelity. Examples of mismatched bases include a G/T or A/C pairing. The damage is repaired by excising the wrongly incorporated base and replacing it with the correct nucleotide. Usually, this involves more than just the mismatched nucleotide itself, and can lead to the removal of significant tracts of DNA. NB MLH1, IP NB MLH1 (164C819) NB MLH1 (Agarose) IP H M02 MLH1 (M1), IHC-P, ELISA NB MLH3 Goat ELISA, NB MSH gamma 3 IHC-Fr Rt NB MSH2, IP NB MSH2 IHC, Mu NB MSH2, Mu H A01 MSH2 NB MSH2, IP NB MSH2, IP, Mu NB MSH2 (25D12) IHC-Fr, IHC-P NB MSH2 (3A2B8C), IHC-P, IHC-Fr, ELISA NB MSH2 (Agarose) IP NB MSH3 Goat, ELISA NB MSH4 Goat, ELISA NB MSH5 Goat, ELISA NB MSH6 Goat, IP, ICC NB MSH6, IP NB MSH6 IHC-P NB MSH6 (44), IP, IHC-P Ca,, Mu, Rt NB PMS1 Goat, ELISA NB PMS2, IP NB PMS2 (163C1251), Mu NB PMS2 (Agarose) IP WANT YOUR ANTIBODY PRODUCED FOR FREE? Visit our website, and fill out the Antibody Grant Sheet for a chance to receive 2 mgs of FREE antibody! Grant Award Date: 1 Award selected on the 15th of every month. Awardees will receive a 0.2 mg test sample of affinity purified rabbit sera. (Typical antibody production takes 4-5 months). If the product works and you supply the necessary documentation, you will receive 2 mgs of affinity purified antibody in exchange for product feedback. Novus reserves the right to sell the antibody produced by the grant. Submit by the end of the month to be selected in the following month s drawing by fax (below) or (novus@novusbio.com). 10

12 DNA Polymerase Antibodies DNA polymerase is an enzyme that assists in DNA replication. Such enzymes catalyze the polymerization of deoxyribonucleotides alongside a DNA strand, which they read and use as a template. The newly-polymerized molecule is complementary to the template strand and identical to the template s partner strand. DNA-Polymerase initiates DNA replication by binding to a piece of single-stranded DNA. NB DNA Polymerase Beta, ELISA NB DNA Polymerase Beta (18S), IP Bv, Ha,, Mu, Rt, Xp NB DNA Polymerase Beta (61) IHC-Fr, IHC-P, NB DNA Polymerase Delta (607) H M01 DNA Polymerase E2 (1A3) ELISA, H B01 DNA Polymerase E3 ELISA, H A01 DNA Polymerase E4 ELISA, NB DNA Polymerase Epsilon (3C5.1), IP (weak) Dr(-), Ha,, Mu, Mk, Ye(-), Ft, Vi(-) NB DNA Polymerase Gamma, IP, ICC, Mu NB DNA Polymerase H IP NB DNA Polymerase Iota NB DNA Polymerase Lambda Goat ELISA, PCNA See Base Excision Repair H A01 REV3L ELISA, DNA Polymerase Beta NB of DNA Polymerase Beta in HeLa whole cell lysate DNA Polymerase Epsilon NB Direct Reversal of Damage Antibodies Direct reversal of O6 adducts caused by chemotherapy agents is accomplished in mammalian cells by the protein O6-methylguanine DNA NB MGMT, Mu NB MGMT (MT 23.2), IHC, Mu(-) of HeLa DNA Polymerase Epsilon in HeLa whole cell lysates methyltransferase (MGMT). Some tumors overexpress MGMT and are resistant to alkylator therapy. 11

13 Longevity Antibodies A number of individual genes have been identified that influence variations in lifespan within a population of organisms. The effects of these genes are strongly dependent on the environment, particularly on the organism s diet. Caloric restriction reproducibly results in extended lifespan in a variety of organisms, likely via nutrient sensing pathways and decreased metabolic rate. The molecular mechanisms by which such restriction results in lengthened lifespan are unclear; however, the behavior of many genes known to be involved in DNA repair is altered under conditions of caloric restriction. Age-1 (PI3 kinase) Akt (tumor suppressor gene) SGK-1 (glucorticoid inducible kinase) Indy (mitochondrial substrate transporter). mutant not as efficient Tert (telomerase reverse transcriptase) Daf-2 (I/IGF receptor) Daf-12 (nuclear hormone receptor) Daf-16 (transcription receptor) HSF-1 (heat shock Sirt2 (histone deacetylase) factor) FOXO3a (forkhead anti-oxidant transcription proteins factor) Ku70 DNA repair (DNA repair oxidant factor) neutralization reduction in oxidant DNA repair production DNA repair HSF1 NB Staining of paraffinembedded human ovarian carcinoma. TERT NB DNA Image courtesy of Harold Brenner. NB DAF-3, CHIP Ce NB FOXO1a [7H3] NB FOXO3A, IF, Mu NB FOXO3A, IP Dr(-), NB FOXO3A, IP Dr(-), NB FOXO3A Goat H M08 FOXO3A [4F2] IHC NB HSF1 [EP1710Y], IP, IF, IHC, FACS, Mu(-), Rt(-) NB HSF1 [SPM401] Rat, IP. IF H R01 HSF1 Pre-design Chimera RNAi Ku70 See Non-Homologous End Joining H M01/M02/ M03 SGK [4D7-G3] IHC H P01/Q01 SGK Recombinant Protein NB SIRT1 IP, NB /2133 SIRT1 IP, NB SIRT2, IF, ICC Mu NB SIR2/SIRT1 [E104], IP. IHC, ICC, Mu(-), Rt(-) NB SIR2/SIRT1 [E54], IP. ICC, IHC, FACS, Mu(-), Rt(-) NB SIRT2, IP NB SIRT2 [EP1668Y], IP, ICC, FACS, Rt H R01 SIRT2 Pre-design Chimera RNAi NB TERT Kit See individual datasheets NB TERT IHC NB TERT, IP. IHC, ICC NB TERT [2C4], IHC, ICC NB TERT [2D8] NB TERT [Y182], ICC, IHC, FACS, Mu(-), Rt(-) H R01 TERT Pre-design Chimera RNAi of human TERT in MJ90 cells. TERT NB TERT NB DAF-3 NB FOXO3A NB Immunofluorescence microscopy of Saos-2 cells transduced with a retroviral vector expressing htert. Staining of exocrine cells and a subset of islets of Langerhans. of DAF-3 in wt c.elegans. Lane 1: wild worm type Lane 2: DAF-3 deletion worms IP/ of human FOXO3A in MCF7 cells. 13

14 Syndromes Linked to DNA Repair Defects Defects in DNA repair are responsible for several genetic disorders, including xeroderma pigmentosum, Cockayne syndrome and trichothiodystrophy. Werner s syndrome, Bloom s syndrome and ataxia telangiectasia are often called accelerated aging diseases because their victims appear elderly and suffer from agingrelated diseases at an abnormally young age. One other group of diseases associated with reduced DNA repair function include Fanconi s anemia, hereditary breast cancer and hereditary colon cancer. Disease Ataxia Telangiectasia ATM See Homologous See HR Recombination Bloom Syndrome NB BLM, IHC NB BLM Goat, IHC NB BLM IHC Mu NB BLM IHC Mu NB BLM, IHC Mu NB BLM (BFL-103), IHC H A01 BLMH, ELISA Hereditary Breast Cancer BRCA1 See Homologous See Recombination Hereditary Breast Cancer BRCA2 See Homologous See Recombination Fanconi Anemia NB FANCA NB FANCC NB FANCD2, IHC, ICC, IF, Mu NB FANCD2 (103), ELISA NB FANCD2 (FI-17), IHC Ha(-),, Mu(-) NB FANCD2 [Ser222] NB FANCG H M01 FANCG (2C8), ELISA NB FANCI, IHC, IP NB FANCJ, IP, IHC NB FANCJ (GO11-3E6) NB FANCJ (pp15-ib4) Hereditary Nonpolyposis Colorectal Cancer MSH2 See Mismatch See MR Repair MLH1 See Mismatch See MR Repair PMS1 See Mismatch See MR Repair PMS2 See Mismatch See MR Repair Werner Syndrome NB WRN, IHC NB WRN, IHC Xeroderma Pigmentosum XPA XPB XPC XPD XPF XPG See Nucleotide Excision Repair See Nucleotide Excision Repair See Nucleotide Excision Repair See Nucleotide Excision Repair See Nucleotide Excision Repair See Nucleotide Excision Repair Can t Decide? Try a SuperNovus Pack: NB BLM (man) SuperNovus Pack NB BLM () SuperNovus Pack SAMPLE SIZES AVAILABLE FANCJ NB WRN NB BLM NB of human BLM in 293T cells. BLM NB IP/ of BLM in NIH3T3 cells. FANCD2 NB FancD2 colocalizes in vivo with another protein in SiHa cells after cell exposure to IR. FANCD2 NB of FANCD2 antibody in HeLa whole cell extract (NB 800-PC1). of BRIP1 in 293 cell lysate. of WRN in 293T whole cell lysate. 13

15 IN THE NEWS 1. [DNA Ligase 1 NB ] Rose, J. L., et al. Base Excision Repair Proteins Are Required for Integrin- Mediated Suppression of Bleomycin-Induced DNA Breakage in Murine Lung Endothelial Cells. J. Pharmacol. Exp. Ther. 321: , [Mre11 NB ] [NBS1 NB ] Kruger, L., et al. Cancer incidence in Nijmegen breakage syndrome is modulated by the amount of a variant NBS protein. Carcinogenesis, 28: , [RAD50 NB ] [Mre11 NB ] Cahill, Dana, et al. Dimerization of the Rad50 protein is independent of the conserved hook domain. 22: , [XRCC3 NB ] [NBS1 NB ] [XRCC3 NB ] Compton, S.A., et al. Xrcc3 and Nbs1 Are Required for the Production of Extrachromosomal Telomeric Circles in man Alternative Lengthening of Telomere Cells. Cancer Res. 67: , [RAD51C NB ] Kuznetsov, S., et al. RAD51C deficiency in mice results in early prophase I arrest in males and sister chromatid separation at metaphase II in females. J. Cell Biol. 176: , [FANCD2 NB ] Jacquemont, C. et. al., Proteasome Function Is Required for DNA Damage Response and Fanconi Anemia Pathway Activation. Cancer Res. 67: , [FANCJ NB ] Kim, J. M., et al. Cell cycle dependent chromatin loading of the fanconi anemia core complex by FANCM/FAAP24. Blood: , [BLM NB ] Tripathi, et al. BLM helicase dependent and independent roles of 53BP1 during replication stress-mediated homologous recombination. J. Cell Biol. 178: 9-14, [BRCA1 NB ] Zaugg, K., et al. Cross-talk between Chk1 and Chk2 in double-mutant thymocytes. PNAS. 104: , [ATM NB ] Dirlam, A. et al. Deregulated E2f-2 underlies cell cycle and maturation defects in Rb null erythroblasts. Mol. Cell. Biol. 27: , [ATM NB ] Nomura, Y. et al. Original Articles: man Mus81 and FANCB independently contribute to repair of DNA damage during replication. Gene Cells. 12: , [ATR NB ] Singh, R., et al. Silibinin suppresses in vivo growth of human prostate carcinoma PC-3 tumor xenograft. J. Nucl. Med. 48(12): , [MSH6 NB ] Masih, P., et al. Mismatch Repair proteins are recruited to replicating DNA through interaction with Proliferating Cell Nuclear Antigen (PCNA). Nucleic Acids Res. 36: 67-75, [ATRIP NB ] Choi, J. et al. Reconstitution of a human ATR-mediated checkpoint response to damaged DNA. PNAS. 104(33): , [WRN NB ] Harrigan J.A., et.al. Metal-catalyzed Oxidation of the Werner Syndrome Protein Causes Loss of Catalytic Activities and Impaired Protein-Protein Interactions. J Biol Chem. 282(50): , [DNA Polymerase Delta NB ] Kundu, C., et al. Adenomatous polyposis coli-mediated hypersensitivity of mouse embryonic fibroblast cell lines to methylmethane sulfonate treatment: implication of base excision repair pathways. Carcinogenesis. 28: , [53BP1 NB ] Murga, M., I. Jaco, et al. (2007). Global chromatin compaction limits the strength of the DNA damage response. 178: [KAP1 NB ] Yang, B. et al. Experimental Therapeutics, Molecular Targets, and Chemical Biology: Mage-A, mmage-b, and MAGE-C Proteins Form Complexes with KAP1 and Suppress p53-dependent Apoptosis in MAGE-Positive Cell Lines. Cancer Res. 67: ,

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