Antibodies. for the study of CELL BIOLOGY & CANCER RESEARCH

Transcription

1 Antibodies for the study of CELL BIOLOGY & CANCER RESEARCH

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3 Antibodies for the study of CELL BIOLOGY & CANCER RESEARCH This pamphlet includes Cayman s most significant antibodies for the identification of kinase signaling, apoptosis, proliferation, factors related to DNA damage/repair, tumor metastasis, and much more. Cayman also provides relevant cell signaling inhibitors, recombinant proteins, assay kits, and other biological reagents. Learn more information about the products we offer in this important research area at Table of Contents 2 PI3 Kinase/Akt 3 Apoptosis 3 Protein Translation/Synthesis 4 ERK/MAPK cascade 4 Guanylate Cyclase 4 Hypoxia 4 DNA Damage and Repair 5 Stem Cell Renewal 5 Cytoskeleton, Motility, and Chemotaxis 5 pan-cytokeratin 5 Cell Cycle/Proliferation 6 Sphingolipids 7 SMAD 7 Telomeric repeat binding factor 7 Ubiquitin 7 Transcription Factors 8 Tumor Related 8 Protein Expression 8 Protein Phosphorylation 8 Heat Shock Proteins Ordering Information Orders are accepted by telephone, fax, mail, , or via the Cayman Chemical website. We will accept telephone orders Monday through Friday from 8 AM to 6 PM EST. All orders received by 1 PM EST will be shipped the same day if stock is available (Monday through Thursday only). Confirming purchase orders must be clearly marked as such to avoid possibility of duplication. Domestic Shipments In most instances we ship FedEx Standard Overnight Delivery (not available to all locations), with delivery by 3:30 PM of the next business day. Product availability may vary. Local delivery is available for the Ann Arbor area only. Other shipping options will be considered upon request, but can be granted only under conditions that will ensure the quality of the product. Freight is prepaid and added to the invoice. Please inquire at the time of order for an estimate of the freight charges. If you wish us to ship collect, please supply a valid account number when ordering. Please address all orders to: Cayman Chemical Company Toll-free Phone: (800) E. Ellsworth Road Fax: (734) Ann Arbor, MI USA custserv@caymanchem.com Phone: (734) Include the following information with your order: 1. Catalog number, description, size, and quantity desired. 2. Complete shipping address. (Delivery is not available to post office box numbers.) 3. A complete billing address. 4. A purchase order number or major credit card (Visa, MasterCard, or American Express), account number, and expiration date. 5. Name of the end user. Terms 1. U.S. funds only, drawn on a U.S. bank. 2. Net 30 days. 3. F.O.B. Ann Arbor, Michigan, U.S.A. 4. Bank fees and wire transfer fees are not to be deducted from the invoice amount. Returns Products cannot be returned without prior authorization from Cayman Chemical Company. Please contact our Customer Service Department for return shipping instructions. Custom orders and radioactive material cannot be accepted for return credit if due to a customer s error. Technical Assistance Technical assistance is available from 8 AM to 5:30 PM EST. If inquiring about a purchased product, please provide the catalog number, lot number, and date of purchase to our technical staff so they may answer your questions quickly. Technical assistance may be reached toll free at , via at techserv@caymanchem.com, or on the web at Use of Research Products The products in this catalog are not for human or veterinary disease diagnosis or therapeutic drug use. They should be used only by technically qualified individuals or those under their direct supervision. Any individual working directly with these products should have free access to the applicable Material Safety Data Sheet (MSDS) and should read and understand it completely prior to use. Please contact our Customer Service Department or visit the specific product page on our website if you require additional copies of any MSDS. Patent Disclaimer The end-user assumes full responsibility for appropriate licensing and/or non-infringement for any proprietary claim or patent. Cayman Chemical Ann Arbor, MI

4 [CELL SIGNALING & CANCER] PI3 Kinase/Akt PINK1 Polyclonal Item No. Item Name 1 66 kda Applications IP, WB H, Mo Akt1 (Phospho-Ser473) Monoclonal (Clone 104A282) PH Domain Leucine-rich Repeat Protein ICC, IHC, WB Phosphatase 1 Polyclonal PHLPP1α and PHLPP1β; (-) PHLPP PTEN Polyclonal IHC, WB PINK1 Polyclonal IHC, WB 33 kda Lane 1: Mouse liver 100,000 x g pellet (30 µg) Akt in Cell Signaling Akt, also known as protein kinase B (PKB), is RTK a major downstream effector of PI3 kinase. Akt is phosphorylated on its activation loop by PDK1 and on its hydrophobic motif by AP PHD Both phosphorylation events are needed for full Akt activity. PIP3 PIP3 PI3K various kinases, including mtor in mtorc2. P PDK1 AKT kinases and transcription factors, producing a wide variety of immediate and delayed GSK3β TSC1/2 Mdm2 Rheb p53 glucose metabolism IKKα by mtorc1 activating AS160 and promoting GLUT4 translocation and glucose uptake. Akt also phosphorylates altering FOXO1 apoptosis, DNA repair, cell cycle arrest, glucose metabolism cell cycling, glucose metabolism effects on cell function. For example, stimulates mtorc2 P AS160 Akt phosphorylates numerous enzymes, Akt PHD and glucose inactivates metabolism GSK3β, and PRAS40 BAD Caspase-9 apoptosis apoptosis, DNA repair, cell cycle arrest cell cycling, protein translation Some targets and processes affected by activated Akt. cell cycling. The phosphorylation of TSC1/2 by Akt prevents it from negatively regulating the small G protein Rheb, allowing accumulation of Rheb and activation of mtorc1. Akt also targets PRAS40, a component of mtorc1. These actions stimulate protein translation and enhance cell growth. Akt prevents apoptosis by acting through different pathways. It activates IKKα, increasing cyclin expression via NF-κB. It phosphorylates BAD, preventing it from heterodimerizing with Bcl-XL/Bcl-2 to initiate apoptosis. In addition, Akt inactivates caspase-9, blocking the caspase cascade leading to apoptosis. The tumor suppressor p53 is a transcription factor which impacts apoptosis, DNA repair, and cell cycle arrest. The phosphorylation of Mdm2 by Akt promotes its nuclear import, leading to the ubiquitination, nuclear export and proteolysis of p53, suppressing its transcriptional activity. The forkhead transcription factor FOXO1 regulates gene expression relevant to a wide variety of cellular functions, including apoptosis, DNA repair, cell cycle arrest, and glucose metabolism. Phosphorylation of FOXO1 by Akt sequesters FOXO1 in the cytoplasm, preventing transcriptional activation. 2 ChIP chromatin immunoprecipitation ELISA enzyme-linked immunosorbent assay FC flow cytometry ICC immunocytochemistry IF immunofluorescence IHC immunohistochemistry IP immunoprecipitation WB western blot

5 Apoptosis [CELL SIGNALING & CANCER] AIF Polyclonal WB Apaf-1 Polyclonal WB H, Mo ARC Polyclonal Ip, WB H, Mo Bim/BOD (IN) Polyclonal IHC, WB Caspase-1 Monoclonal (Clone 14F468) IHC, WB H, Mo Caspase-3 (human) Polyclonal WB, IHC H, Mo, Bb Caspase-3 Monoclonal (Clone 31A1067) IHC, WB Caspase-3 Monoclonal (Clone 31A893) WB H Caspase-8 Monoclonal (Clone 90A992) FC, IHC, WB H, RMk, Chimp Caspase-9 (carboxy-terminal divergent) Polyclonal Caspase-9 Polyclonal WB H WB Caspase-14 Monoclonal (Clone 70A1426) FC, WB H, Mo DcR2 Polyclonal WB H EAF2 Polyclonal IHC, WB Programmed Cell Death Protein 4 (C-Term) Polyclonal WB RICK Polyclonal WB not available IGFBP5 Polyclonal WB Mo, R IP6K2 Monoclonal (Clone 4F10) ICC, WB H, Mo TRAIL Polyclonal WB H, Mo Protein Translation/Synthesis eif3g Polyclonal (aa ) IHC, WB Chimp, Bo, C, Ho,, RMk Bb baboon Be beluga Bo bovine C canine CE C. elegans Ch chicken Chimp chimpanzee Co coral DM drosophila EC E. coli GP guinea pig H human Ha hamster Ho horse Mk monkey Mo mouse Mq macaque NWMk new world monkey Or orangutan P pig R rat Rb rabbit RMk rhesus monkey Sh sheep X xenopus 3

6 [CELL SIGNALING & CANCER] ERK/MAPK cascade ERK/MAPK (Phospho-Thr 202 /Tyr 204 ) Polyclonal WB H, R MEK1 (Phospho-Thr 292 ) Polyclonal WB MEK1 (Phospho-Thr 386 ) Polyclonal WB MEK1/2 (Phospho-Ser 218,222 ) Polyclonal WB NIH 3T3 cells p38 MAPK (Phospho-Thr 180 /Tyr 182 ) Polyclonal WB H p38 MAPK Monoclonal (Clone 9F12) FC, ICC, WB H, R; Bo, C, Ch, Mo, X (expected); Non-human primates ME 1 (expected) R; Bo, C, Ch, Mo, Non-human primate, X (expected) Raf-1 (Phospho-Ser 301 ) Polyclonal WB Raf-1 (Phospho-Ser 642 ) Polyclonal WB R; Bo, Ca, Ch, H, Mo, Non-human primate, X (expected) R; Bo, C, Ch, H, Mo, Non-human primate, X (expected) Ribosomal S6 Kinase 2 Polyclonal WB R; Bo, C, Ch, Mo, H (expected) p53 Monoclonal (Clone BP53-12) 250 kda 150 kda 100 kda kda 110 kda Guanylate Cyclase Guanylate Cyclase α subunit (soluble) Polyclonal Guanylate Cyclase β1 subunit (soluble) Polyclonal WB IHC, WB H, Mo, Bo Most Mammilian Species 75 kda 50 kda 37 kda 25 kda Lane 1: Cos cell lysate (30 µg) Lane 2: Cos cell lysate (40 µg) Lane 3: CoCl2 treated Cos cell lysate (30 µg) Hypoxia HIF-1α (C-Term) Polyclonal WB H, Mo, Simian HIF-1α Monoclonal (Clone H1α67) IHC, WB Ferret, H, Mo, Sh DNA Damage and Repair PARP (Cleaved) Monoclonal FC, WB H p53 (Phospho-Ser392 ) Polyclonal p53 Monoclonal (Clone BP53-12) WB FC, ICC, IHC, WB H p53 Expression in non-hodgkin s lymphoma (immunoperoxidase method) 4 ChIP chromatin immunoprecipitation ELISA enzyme-linked immunosorbent assay FC flow cytometry ICC immunocytochemistry IF immunofluorescence IHC immunohistochemistry IP immunoprecipitation WB western blot

7 Phospholipase [CELL SIGNALING & CANCER] LPA1 Polyclonal LPA1 Polyclonal ICC, WB Bo,, Sh LPA3 Polyclonal ICC, WB Lysophospholipase D Polyclonal ICC, WB NAPE-PLD Polyclonal (aa 6-20) WB, Bo NAPE-PLD Polyclonal (aa ) WB Cytoskeleton, Motility, and Chemotaxis Immunofluorescent staining of A549 cells with Cayman s LPA 1 polyclonal antibody (8 mg/ml). The positive reaction is visualized by a Cy3-conjugated donkey anti-rabbit secondary antibody Lysophospholipase D Polyclonal ARAP2 Polyclonal IHC, WB H Chemokine-Like Receptor 1 Polyclonal FC, ICC, WB H, Mk, Mo, R VE-Cadherin Polyclonal ICC, IP, WB H, Bo, Mo Podoplanin Monoclonal (Clone pmab-1) Podoplanin Monoclonal (Clone NZ-1.2) FC, WB FC, WB M H pan-cytokeratin Cytokeratin Monoclonal (Clone C-11) Cytokeratin Monoclonal APC (Clone C-11) Cytokeratin Monoclonal FITC (Clone C-11) Cytokeratin Monoclonal PE (Clone C-11) FC, ICC, IHC, WB FC FC, IF FC, IF H cytokeratin 4, 5, 6, 8, 10, 13, 18 H cytokeratin 4, 5, 6, 8, 10, 13, 19 H cytokeratin 4, 5, 6, 8, 10, 13, 20 H cytokeratin 4, 5, 6, 8, 10, 13, 21 Immunofluorescent staining of HepG2 cells with the lysopld polyclonal antibody (2 mg/ml). The positive reaction is visualized by a Cy3-conjugated donkey anti-rabbit secondary antibody. [NOTE: the cytoplasmic and membrane localization of the protein Cytokeratin Monoclonal (Clone C-11) Cell Cycle/Proliferation PCNA Monoclonal (Clone IPO-38) FC, ICC, IHC, WB Multiple species Human cervical carcinoma cells stained with Cytokeratin Monoclonal (1 μg/ml) followed by goat anti-mouse IgG-Cy Cytokeratin Monoclonal PE (Clone C-11) HeLa cells stained with Cytokeratin Monoclonal PE (Clone C-11), 10 µg/ml. Bb baboon Be beluga Bo bovine C canine CE C. elegans Ch chicken Chimp chimpanzee Co coral DM drosophila EC E. coli GP guinea pig H human Ha hamster Ho horse Mk monkey Mo mouse Mq macaque NWMk new world monkey Or orangutan P pig R rat Rb rabbit RMk rhesus monkey Sh sheep X xenopus 5

8 [CELL SIGNALING & CANCER] Sphingolipids S1P1 Polyclonal kda Lane 1: Human liver microsomes (15 µg) Lane 2: Murine brain homogenate (30 µg) Lane 3: Porcine liver 10,000 x g supernatant (30 µg) Lane 4: Rat brain homogenate (30 µg) Item No. Item Name Applications S1P1 Polyclonal ICC, IHC, WB H, Mo, P, R WB H, Mk, Mo, R S1P3 Polyclonal ICC, WB S1P4 Polyclonal ICC, IF, WB H, Mo S1P5 Polyclonal IHC, WB H, C, Mk, Mo, R Serine Palmitoyltransferase Polyclonal IHC, WB, Sh, P, Bo, African green Mk Sphingosine Kinase 1 Polyclonal ICC, WB H, Mo; R (expected) Sphingosine Kinase 1 Polyclonal FITC FC, IF, WB Mo, H; R (expected) S1P2 Polyclonal From Ceramide to Sphingosine 1-phosphate and S1P Receptors OH OH N H Ceramide occurs in abundance in the outer leaflet of the cell membrane. It is synthesized de novo by the serine palmitoyl transferase pathway, starting with serine and O Ceramide palmitate. ceramidase Ceramide can also be rapidly released from sphingomyelin by OH OH sphingomyelinases. Consisting of sphingosine joined with a fatty acid of variable length, ceramide is enzymatically cleaved to give these components by acid and neutral ceramidases. Liberated sphingosine can then be phosphorylated to produce sphingosine 1-phosphate (S1P) by two distinct sphingosine kinases, NH2 Sphingosine Sphingosine kinase Sphingosine phosphatase O OH SPHK1 and SPHK2. SPHK1, the better studied form, is activated by O _ O many stimuli, including TGF-β, IL-1β, TNF-α, PDGF, HGF, LPS, RSV, insulin, and BCR/ABL. SPHK1 is best known as a survival, or antiapoptosis, enzyme, with additional positive effects on cell motility and proliferation, resulting from the production of S1P. These _ O P Sphingosine-1-phosphate NH2 Sphingosine, derived from ceramide, is phosphorylated by sphingosine kinases. actions contribute to tumor growth and metastasis. In addition, SPHK1-derived S1P activates endothelium, regulating endothelial barrier homeostasis, primes neutrophils, activates macrophages and promotes phagosome maturation, and increases immune cell motility and function. S1P signals through G-protein coupled receptors. These receptors were initially identified as endothelial differentiation gene (EDG) receptors. With the identification of S1P as a ligand for five of the EDG receptors, they have been renamed: SIP1 (EDG1), S1P2 (EDG5), SIP3 (EDG3), SIP4 (EDG6), and SIP5 (EDG8). Cells overexpressing SIP1 and SIP3 have been shown to signal primarily through Gi, inhibiting adenylate cyclase-mediated increase of camp. Commonly, the main role of S1P2 is to oppose or balance the signaling of SIP1 through its G12-mediated activation of Rho, which inhibits Rac. SIP4 and SIP5 have been less well studied. In lymphocytes, activation of Cdc42 via SIP4 contributes to S1P-mediated chemotaxis. Both SIP4 and SIP5 also suppress Rac via G12/13 activated Rho. 6 ChIP chromatin immunoprecipitation ELISA enzyme-linked immunosorbent assay FC flow cytometry ICC immunocytochemistry IF immunofluorescence IHC immunohistochemistry IP immunoprecipitation WB western blot

9 SMAD [CELL SIGNALING & CANCER] SMAD1/5/8/9 Polyclonal WB SMAD2 Polyclonal IHC, WB Bo, C, Ch, Chimp, SMAD3 Polyclonal IHC, WB Bo, C, H, Mo, P, R, RMk SMAD4 Polyclonal WB H, NWMk, Mo, R SMAD6 Polyclonal IHC, WB H, NWMk, Mo, Sh, R SMAD7 Polyclonal WB, Sh SMAD7 Polyclonal (azide-free) WB Telomeric repeat binding factor TRF2 Monoclonal (Clone 4A794.15) FC, ICC, WB H, Mo, Muntjac, R TRF2 Polyclonal ChIP, ICC, IP, WB H, Mo Ubiquitin Multiubiquitin Chain Monoclonal (Clone FK1) ELISA, WB Does not react with methyl ubiquitinized protein and free-ubiquitin Multiubiquitin Chain Monoclonal (Clone FK2) ELISA, WB Reacts with methyl ubiquitinized protein SUMO Polyclonal ELISA, WB not available SUMO Monoclonal ELISA, WB not available Ubiquitin Monoclonal (Clone 5B9-B3) ELISA, WB, Bo Ubiquitin Monoclonal (Clone 6C11-B3) ELISA, WB, Bo Ubiquitin Polyclonal ChIP, IP, WB H, Mk, Mo, R, Ha, Rb, GP, Bo, P, C, Sh, Ch, X, Yeast, DM, Rainbow Trout Transcription Factors ATF2 (Phospho-Ser 490,498 ) Polyclonal IHC, WB H Bnc 1 Polyclonal WB Chimp, H, Mk CREB (Phospho-Ser 133 ) Polyclonal WB R Bb baboon Be beluga Bo bovine C canine CE C. elegans Ch chicken Chimp chimpanzee Co coral DM drosophila EC E. coli GP guinea pig H human Ha hamster Ho horse Mk monkey Mo mouse Mq macaque NWMk new world monkey Or orangutan P pig R rat Rb rabbit RMk rhesus monkey Sh sheep X xenopus 7

10 [CELL SIGNALING & CANCER] Hepsin Polyclonal (aa ) Tumor Related Hepsin Polyclonal (aa )IHC, WB H, Mo SUCNR1 Polyclonal IHC, WB H, Chimp, NWMk Protein Expression DYKDDDDK Polyclonal WB not available Top Panel: 10 µg/ml Hespin Polyclonal (Item No ) followed by anti-rabbit secondary with NovaRed and finally hematoxylin stain development HA Polyclonal WB not available Protein Phosphorylation Phosphothreonine (Ascites) Monoclonal (Clone 18F6) ELISA, WB not available Phosphotyrosine (Ascites) Monoclonal (Clone 13F9) ELISA, WB not available Bottom Panel: Anti-rabbit secondary alone with NovaRed and hematoxylin stain development steps. Heat Shock Proteins Hsc70 (Hsp73) Polyclonal IF, IP, WB H, Ha, R Hsf1 Monoclonal (Clone 10H8) ELISA, ICC, IP, WB, Bo, GP, Ha, Mk, Rb Hsf2 Monoclonal (Clone 3E2) WB, GP, Ha, Mk, Rb, C, Bo, Sh, P Hsp25/Hsp27 Monoclonal (Clone 8A7) ICC, IF, IP, WB Mo, R, Bo, C, GP, Ha, H Hsp40 Polyclonal WB H, R Hsp60 Monoclonal (Clone LK-1) ELISA, FC, IP, WB Hsp60 Monoclonal (Clone LK-2) FC, IHC, WB Hsp70 (Hsc70) Monoclonal (Clone BB70) IHC, IP, WB Hsp70 (Hsc70) Monoclonal (Clone N27) FC, IHC, IP, WB Hsp70 Polyclonal WB, IP, ELISA, IHC, ICC, Bo, C, Ch, DM, GP, Ha, Mk, P, Rb, Sh, X, Bo, C, Ch, GP, Ha, Mk, P, Rb, Spinach, EC, H. pylori, S. typhimurium, T. spiralis, Yeast, White fly, Sh, C, Be, Bo, Fish (carp, rainbow trout, chinook/chum salmon), GP, Ha, Rb, Ch, X, DM, Yeast, Bo, CE, Be, C, Ch, DM, Fish (carp, Chinook/chum salmon, rainbow trout), GP, Ha, Mk, P, Plant (cucumber/ pea), Rb, Sh, X, Be, Bo, C, Carp, GP, Mk, P, Sh, coral 8 ChIP chromatin immunoprecipitation ELISA enzyme-linked immunosorbent assay FC flow cytometry ICC immunocytochemistry IF immunofluorescence IHC immunohistochemistry IP immunoprecipitation WB western blot

11 [CELL SIGNALING & CANCER] Heat Shock Proteins cont Hsp70 Monoclonal (Clone C92F3A-5) ELISA, ICC, IHC, IP, WB Hsp70 Monoclonal FITC (Clone C92) FC, Bo, CE, C, Ch, DM, Fish (carp), GP, Ha, Mk, P, Rb, Sh; (-) Hsc70(Hsp73) Hsp70, Bo, C, Ch, DM, Fish (carp), GP, Ha, Mk, P, Rb, Sh Hsp90 Polyclonal Antiserum WB, ELISA, IP H, R, Mo Hsp90 Monoclonal (Clone AC-16) WB H, Rb, R, Mo, Ch, Achlya, Wheat germ, Sƒ9 cells; (-) EC, yeast Hsp90 Monoclonal Complex (Clone 8D3) IP, Rb Hsp90α Monoclonal (Clone D7α) IP, WB Bo, Ch, H, Mo, P, R, Rb Hsp90; H Hsp90α Hsp90α Monoclonal (Clone K41009) ELISA, IHC, WB H (α-specific) Hsp90, R Hsp90β Hsp90α/β Monoclonal (Clone K41220A) ELISA, WB H (β-specific), R, S. cerevisiae, S. pombe, Rice, P. caudatum Hsp90β Monoclonal (Clone H90-10) ELISA, IP, WB H, Rb, Ch, R Hsp90β Monoclonal (Clone K3701) ELISA, IHC, WB H (β-specific) Hsp90; (-) H Hsp90α, R Hsp Hsp90β Polyclonal ELISA, IP, WB H, R, Mo Wnt Signaling PORCN Polyclonal FC, IF, WB H β-catenin Polyclonal IHC, WB Bo, H, R, Mo, P β-catenin (Phospho-Ser33,37) Polyclonal WB H Bb baboon Be beluga Bo bovine C canine CE C. elegans Ch chicken Chimp chimpanzee Co coral DM drosophila EC E. coli GP guinea pig H human Ha hamster Ho horse Mk monkey Mo mouse Mq macaque NWMk new world monkey Or orangutan P pig R rat Rb rabbit RMk rhesus monkey Sh sheep X xenopus 9

12 Cayman Chemical Ann Arbor, MI