APOA1 Antibody (OASA09127)

Catalog#: OASA09127

• Size: 0.5mg

• Description of Target: SHEEP ANTI HUMAN APOLIPOPROTEIN A1
• Gene Symbol: APOA1
• Alias Symbols: MGC117399; APOA1
• Protein Accession#: NP_000030.1
• Swissprot Id: P02647
• Host: Sheep
• Protein Size (# AA): 267
• Specificity: APOLIPOPROTEIN A1
• Application: ELISA
• Immunogen: The immunogen for anti-APOA1 antibody: purified human apolipoprotein A1
• Product Format: Purified IgG - lyophilised
• Isotype: Polyclonal IgG
• Predicted Homology Based on Immunogen Sequence: Human
• Applications Info: ELISA : This antibody can be used in a sandwich ELISA with OASA00955 as the capture antibody.
  Preparation: Purified IgG prepared by affinity chromatography on immobilized human apolipoprotein A1
  
  Preservative Stabilisers: Contains mannitol, dextran & salts
• Protocol Information: Citation: 1: Sonoyama K, Nishikawa H, Kiriyama S, Niki R. Apolipoprotein mRNA in liver and intestine of rats is affected by dietary beet fiber or cholestyramine. J Nutr.1995 Jan;125(1):13-9. PubMed PMID: 7815170.
  Species: Rat
  Experiment Name: Immunoblotting for plasma apo A-I quantification
  Experiment Background: In this study, Kei et al. have collected data from the liver and examined the effects of diets containing beet fiber, cholestyramine and no fiber (fiber-free) on plasma and liver lipids and tissue apolipoprotein mRNA.
  Experimental Steps: Whole plasma (0.5 µL) was subjected to 12% SDS-PAGE under reducing conditions (Laemmli 1970) and then electrophoretically transferred to nitrocellulose membrane (Hybond C extra, Amersham Inter national, Amersham, U.K.) in a semidry electroblotting apparatus (Nippon Eido, Tokyo, Japan) at constant current of 170 mA for 1 h. The blotting buffer contained 125 mmol/L Tris, 960 mmol/L glycine in 200 mL/L methanol, pH 8.3. The membrane was then incubated in a blocking solution of 50 g/L bovine serum albumin (fraction V, Miles Inc., Kankakee, IL) in Tris-buffered saline (20 mmol/L Tris-HC1, 150 mmol/L NaCl, pH 7.4), followed by the incubation for 1 h with a 1:100 dilution of the sheep anti-human apo A-I serum (Oxford, U.K.) in the blocking solution containing 500 mg/L Tween 20. The membrane was then washed three times with Tris-buffered saline containing 500 mg/L Tween 20. Next, the membrane was incubated for 1 h with a 1:2000 dilution of a second antibody of rabbit antisheep IgG conjugated with horseradish peroxidase (Jackson Immunoresearch Laboratories, West Grove, PA) in the same solution used in the incubation with the first antibody, followed by washing in the sameway with the first antibody. Identification of the antigen-antibody complex was performed using Renaissance Western Blot Chemiluminescence Reagent (DuPont NEN Research Products, Boston, MA) as recommended by the manufacturer. The relative quantity of apo A-I was estimated by scanning densitometry (Dual-Wavelength Flying-Spot Scanner CS-9000, Shimadzu, Kyoto, Japan).
  Number Of Protocols: 1
• Reconstitution and Storage: Reconstitution: Use sterile DI water
  Storage: Store at +4^oC or at -20^oC if preferred.
  Storage in frost-free freezers is not recommended.
  This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody.

Product Protocol: APOA1 antibody used to evaluate apolipoprotein mrna in liver and intestine of rats is affected by dietary beet fiber or cholestyramine (OASA09127)

Specificity: APOLIPOPROTEIN A1
Product Page: APOA1 antibody (OASA09127)
Antibody Pair Available: OASA00955 Capture OASA09127 Detection
Experiment Type: Immunoblotting for plasma apo A-I quantification.

Protocol:
Whole plasma (0.5 uL) as subjected to 12% SDS-PAGE under reducing conditions (Laemmli 1970) and then electrophoretically transferred to nitrocellulose membrane (Hybond C extra, Amersham International, Amersham, U.K.) in a semidry electroblotting apparatus (Nippon Eido, Tokyo, Japan) at constant current of 170 mA for 1 h. The blotting buffer contained 125 mmol/L Tris, 960 mmol/Lglycine in 200 mL/L methanol, pH 8.3. The membrane was then incubated in a blocking solution of 50 g/L bovine serum albumin (fraction V, Miles Inc., Kankakee, IL) in Tris-buffered saline (20 mmol/L Tris-HC1, 150 mmol/L NaCl, pH 7.4), followed by the incubation for 1 h with a 1:100 dilution of the sheep anti-human apo A-I serum (Serotec, Oxford, U.K.) in the blocking solution containing 500 mg/L Tween 20. The membrane was then washed three times with Tris-buffered saline containing 500 mg/L Tween 20. Next, the membrane was incubated for l h with a 1:2000 dilution of a second antibody of rabbit antisheep IgG conjugated with horseradish peroxidase (Jackson Immunoresearch Laboratories, West Grove, PA) in the same solution used in the incubation with the first antibody, followed by washing in the sameway with the first antibody. Identification of the antigen-antibody complex was performed using Renaissance Western Blot Chemiluminescence Reagent (DuPont NEN Research Products, Boston, MA) as recommended by the manufacturer. The relative quantity of apo A-I was estimated by scanning densitometry (Dual-Wavelength Flying-Spot Scanner CS-9000, Shimadzu, Kyoto, Japan).

Summary:
1. The result shows the relative concentrations of plasma apo A-I in rats fed the fiber-free, beet fiber and cholestyramine diets as estimated by immunoblotting analysis. The values in rats fed the beet fiber and cholestyramine diets were expressed relative to the average value in animals fed the fiber-free diet, which was normalized to 100. 2. Plasma apo A-I concentra tions in rats fed the beet fiber diet tended (P < 0.1 ) to be lower than in those fed fiber-free and cholestyramine diets.

References:
1: Sonoyama K, Nishikawa H, Kiriyama S, Niki R. Apolipoprotein mRNA in liver and intestine of rats is affected by dietary beet fiber or cholestyramine. J Nutr. 1995 Jan;125(1):13-9. PubMed PMID: 7815170.