PROCESSING INSTRUCTIONS FOR BRAC TESTS. PROCESSING SPECIFICATIONS Storage 5. Centrifuge Temp ( C) 10 mins. Alternate Draw Tube.

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1 Preferred Amount of Blood (ml) Amount of Blood (ml) PROCESSING INSTRUCTIONS FOR BRAC TESTS Preferred Draw PROCESSING SPECIFICATIONS Storage 5 Alternate Draw Centrifuge 10 mins Preferred Aliquot (ml) Aliquot (ml) 4 ACTH 4 2 EDTA tube on ice none x g 1 2 x x ml 'o' ring -80 Storage s ADIPONECTIN 2 1 SST 1500 x g 1 1 x X ml 'o' ring -80 ALDOSTERONE 2 1 SST 1500 x g 1 1 x 1 1 x ml 'o' ring -80 ANGIOTENSIN II EDTA none x g 1 2 x 1.2 Special 1 x ml 'o' ring 2, 3 Bone Specific Alk Phas (BSAP) 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 C-PEPTIDE 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 Catecholamines, plasma Epinephrine/Norepinephrine 4 2 EDTA none x g 1 2 x 0.8 on ice 1 x 0.8 on ice 2 ml 'o' ring -80 Cortisol 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 C-Terminal telopeptide (CTX) 2 1 SST 1500 x g 1 1 x x ml 'o' ring g = (1118 x 10-8) (radius in cm) (rpm) 2 2 Quick freeze in dry ice/alcohol 3 Special Inhibitor added refer to specific SOP #225 4 Minimum volumes, no repeat, tube will be discarded after analysis is completed 5 Store at -80 C until analysis is performed(samples will be returned to investigator after analysis is completed no later than 4 weeks or didcarded) 7 10 ml is minimum volume recommended per study Revised February 2013 BLOODS 1 of 8

2 PROCESSING INSTRUCTIONS FOR BRAC TESTS PROCESSING SPECIFICATIONS Storage 5 Preferred Amount of Blood (ml) Amount of Blood (ml) Preferred Draw Alternate Draw Centrifuge 10 mins Preferred Aliquot (ml) Aliquot (ml) 4 Storage s DHEAs 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 DNA-extraction (blood) EDTA ACD (acid citrate dextrose), Paxgene or To collect buffy coat spin down at 1500 x g 1 10 mins and remove plasma 1 x 10 ml of blood 1 x 5 ml of blood or 1mL buffy coat Blood must be transferred in sterile 50 ml Falcon tube 50 ml falcon tube must be frozen immediately for both whole blood and buffy coat and store at -80 C E-Selectin 2 1 SST or Citrate 1500 x g 1 1 x X ml 'o' ring -80 Estradiol 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 FSH 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 Grhelin (active) 2 1 EDTA SST 1500 x g 1 1 x x 0.3 Special 2 ml 'o' ring 3-80 Growth Hormone 3 1 SST 1500 x g 1 1 x x ml 'o' ring -80 ICAM 2 1 SST IGF SST 1500 x g 1 1 x X ml 'o' ring x g 1 1 x x ml 'o' ring -80 IL SST 1500 x g 1 1 x x ml 'o' ring -80 Insulin 2 1 SST EDTA 1500 x g 1 1 x x ml 'o' ring -80 Leptin 2 1 SST EDTA 1500 x g 1 1 x x ml 'o' ring g = (1118 x 10-8) (radius in cm) (rpm) 2 2 Quick freeze in dry ice/alcohol 3 Special Inhibitor added refer to specific SOP #225 4 Minimum volumes, no repeat, tube will be discarded after analysis is completed 5 Store at -80 C until analysis is performed(samples will be returned to investigator after analysis is completed no later than 4 weeks or didcarded) 7 10 ml is minimum volume recommended per study Revised February 2013 BLOODS 2 of 8

3 PROCESSING INSTRUCTIONS FOR BRAC TESTS PROCESSING SPECIFICATIONS Storage 5 Preferred Amount of Blood (ml) Amount of Blood (ml) Preferred Draw Alternate Draw Centrifuge 10 mins Preferred Aliquot (ml) Aliquot (ml) 4 Storage s LH 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 Osteocalcin 2 1 SST PAI EDTA or Citrate 1500 x g 1 2x x 0.1 Special 2 ml 'o' ring x g 1 1 x 1 1 x ml 'o' ring -80 Progesterone 2 1 SST none 1500 x g 1 1 x x ml 'o' ring -80 Prolactin 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 PINP 2 1 SST none 1500 x g 1 1 x x ml 'o' ring -80 PTH-Intact 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 PYY (Total) 2 1 SST EDTA 1500 x g 1 1 x X ml 'o' ring 3-80 Renin Activity, Plasma EDTA none 1500 x g 1 2 x 1 Q.F. 2 1 x 1 Q.F. 2 2 ml 'o' ring -80 Active renin 4 2 EDTA none 1500 x g 1 2 x 1 Q.F. 2 1 x 0.5 Q.F. 2 2 ml 'o' ring -80 Resistin 0 1 SST Retinol Binding Protein -4 (RBP4) 2 1 SST 1500 x g 1 1 x x ml 'o' ring x g 1 1 x x ml 'o' ring g = (1118 x 10-8) (radius in cm) (rpm) 2 2 Quick freeze in dry ice/alcohol 3 Special Inhibitor added refer to specific SOP #225 4 Minimum volumes, no repeat, tube will be discarded after analysis is completed 5 Store at -80 C until analysis is performed(samples will be returned to investigator after analysis is completed no later than 4 weeks or didcarded) 7 10 ml is minimum volume recommended per study Revised February 2013 BLOODS 3 of 8

4 PROCESSING INSTRUCTIONS FOR BRAC TESTS PROCESSING SPECIFICATIONS Storage 5 Preferred Amount of Blood (ml) Amount of Blood (ml) Preferred Draw Alternate Draw Centrifuge 10 mins Preferred Aliquot (ml) Aliquot (ml) 4 Storage s Sex Hormone Binding Globulin (SHBG) 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 svcam 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 Testosterone 2 1 SST 1500 x g 1 1 x x ml 'o' ring -80 TNF-α 2 1 SST 1500 x g 1 1 x 1 1 x ml 'o' ring -80 TSH 2 1 SST 1500 x g 1 1 x x ml 'o' ring OH Vitamin D 2 1 SST 1500 x g 1 1 x x ml 'o' ring g = (1118 x 10-8) (radius in cm) (rpm) 2 2 Quick freeze in dry ice/alcohol 3 Special Inhibitor added refer to specific SOP #225 4 Minimum volumes, no repeat, tube will be discarded after analysis is completed 5 Store at -80 C until analysis is performed(samples will be returned to investigator after analysis is completed no later than 4 weeks or didcarded) 7 10 ml is minimum volume recommended per study Revised February 2013 BLOODS 4 of 8

5 ACTH ADIPONECTIN Reportable Units ug/ml Manufacturer Method Lowest REFERENCE RANGE 6 Reportable Value Diasorin, Inc Stillwater, MN IRMA pg/ml ALPCO Diagnostics Salem, NH EIA 0.10 ug/ml Unknown Supine Low Na: ng/dl ALDOSTERONE ng/dl Siemens Los Angeles, CA Coat-A-Count 2.5 ng/dl Upright High Na: 1-10 ng/dl Low Na: ng/dl High Na: 6-20 ng/dl ANGIOTENSIN II ALPCO Diagnostics Salem, NH Extracted 0.6 Supine Low Na: pg/ml High Na: <30 pg/ml Bone Specific Alk Phas (BSAP) ug/l 0.1 ug/l Female Female 56+ Male ug/l ug/l ug/l C-PEPTIDE Seimens Los Angeles, CA Supine: pg/ml Norepi 40 Norepi Standing: pg/ml Catecholamines, plasma Epinephrine/Norepinephrine IBL America Extracted Exercise: pg/ml Supine: pg/ml Epi 1 Epi Standing: pg/ml Exercise: pg/ml Cortisol ug/dl 0.4 ug/dl AM: ug/dl PM.: <10 ug/dl C-Terminal telopeptide (CTX) Immunodiagnostic Systems (IDS) Fountain Hills, AZ 0.02 Post-menopausal Women Pre-menopausal Woman Men For reference only not to be used for diagnostic Revised February 2013 BLOODS 5 of 8

6 DHEAs Reportable Units ug/dl Manufacturer Method Lowest Reportable Value 2 ug/dl REFERENCE RANGE 6 Normal range is gender and age dependant. If you need the normal range please contact the lab. DNA-extraction (blood) E-Selectin ug/ml GENTRA PureGene DNA Isolation 2 ug ug/5-10ml for blood samples that have been stored under optimal conditions. For example, no more than 1 day at room temperature (15 25 C) or 5 days at 4 C or frozen at 80 C for less than 2 years Estradiol 20 Females Non-pregnant -mid-follicular Phase mid-luteal Phase: peri-ovulatory phase Post-menopausal <20-40 Males <20-47 FSH Grhelin (active) Growth Hormone ICAM IGF-1 IL-6 Insulin Leptin miu/ml uiu/ml LINCO Research St. Charles, MS Immunodiagnostic Systems (IDS) Fountain Hills, AZ LINCO Research St. Charles, MS 0.2 miu/ml EIA 2.3 age dependant 0.5 <50 years old: >50 years old: uiu/ml Fasting: uiu/ml 0.1 Females: Follicular Phase miu/ml Mid Cycle Peak miu/ml Mid Luteal Phase miu/ml Post-Menopausal miu/ml Males miu/ml Not Available Males Females Males Females For reference only not to be used for diagnostic Revised February 2013 BLOODS 6 of 8

7 Reportable Units Manufacturer Method Lowest Reportable Value REFERENCE RANGE 6 LH miu/ml 0.2 miu/ml Females: Folicular Phase miu/ml Mid Cycle Peak miu/ml Mid Luteal Phase miu/ml Post-Menopausal miu/ml Males: miu/ml Osteocalcin ALPCO Diagnostics Salem, NH PAI Progesterone 0.08 Females: Follicular Luteal: Post menopausal < Males Prolactin PINP PTH-Intact ug/l Immunodiagnostic Systems (IDS) Fountain Hills, AZ ug/l 1 Post-menopausal Women Pre-menopausal Woman Men Post-menopausal Women ug/l Pre-menopausal Woman ug/l Men ug/l PYY (Total) Millipore St. Charles, MS 10 Unknown Supine High Na: ng/ml/hr Renin Activity, Plasma /hr Diasorin, Inc Stillwater, MN 0.1 /hr Upright Low Na: ng/ml/hr Low Na: ng/ml/hr High Na: ng/ml/hr Active renin uiu/ml IBL America 2 uiu/ml Diet, Posture Dependent - uiu/ml Resistin 0.21 Serum Retinol Binding Protein -4 (RBP4) mg/l ALPCO Diagnostics Salem, NH 1 mg/l Adults mg/l Newborns mg/l 6 months old mg/l 6 For reference only not to be used for diagnostic Revised February 2013 BLOODS 7 of 8

8 Sex Hormone Binding Globulin (SHBG) svcam Testosterone TNF-α TSH 25-OH Vitamin D Reportable Units nmol/l ng/dl uiu/ml Manufacturer Method Lowest Reportable Value Diasorin, Inc Stillwater, MN 0.33 nmol/l REFERENCE RANGE ng/dl 0.5 Males nmol/l Females nmol/l Post-menopausal females nmol/l Male ng/dl Females <10-75 ng/dl uiu/ml uiu/ml 1.5 ng/dl Seasonal/ Regional: ng/ml 6 For reference only not to be used for diagnostic Revised February 2013 BLOODS 8 of 8

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