GOAT ANTI HEPATITIS B SURFACE ANTIGEN AD/AY:HRP Antibody (1ml)

Catalog#: OASA05975

• Size: 1ml

• Description of Target: GOAT ANTI HEPATITIS B SURFACE ANTIGEN AD/AY:HRP
• Alias Symbols: N/A
• Host: Goat
• Specificity: HEPATITIS B SURFACE ANTIGEN AD/AY
• Application: ELISA
• Conjugation: HRP
• Immunogen: Hepatitis B Surface Antigen Subtypes AD & AY
• Product Format: Purified IgG conjugated to Horseradish Peroxidase (HRP) - liquid
• Isotype: Polyclonal IgG
• Applications Info: ELISA : This product may be used in a direct ELISA or as a detection antibody in a sandwich ELISA together with OASA05974 as a capture antibody, and recombinant Hepatitis B surface antigen AY OPSA10763 as a standard.
  Preservative Stabilisers: 0.002% - Thiomersal
  1% - Bovine Serum Albumin
  Approx Protein Conc: IgG concentration 2.0 mg/ml
  
  Buffer Solutions: Phosphate buffered saline, pH 7.2
• Reconstitution and 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.
  Should this product contain a precipitate we recommend microcentrifugation before use.

Product Protocol: GOAT ANTI HEPATITIS B SURFACE ANTIGEN AD/AY:HRP antibody used to evaluate development and technical and clinical validation of a quantitative enzyme-linked immunosorbent assay for the detection of human antibodies to hepatitis b surface antigen in recipients of recombinant hepatitis b virus vaccine (OASA05975)

Specificity: HEPATITIS B SURFACE ANTIGEN AD/AY
Product Page: GOAT ANTI HEPATITIS B SURFACE ANTIGEN AD/AY:HRP antibody (OASA05975)
Antibody Pair Available: OASA05974 Capture OASA05975 Detection
Experiment Type: ELISA procedure

Protocol:
Fifty microliters each of standards, controls, and specimen were added in duplicate to the precoated microplate. The plate was covered and incubated overnight at room temperature and then washed four times with 350 μl of distilled water containing Tween 20 (0.05%) and NaCl (0.154 M). One hundred microliters of HBsAg-HRP conjugate at a working dilution of 1:1,000 in the HRP diluent (0.0095 M PBS [pH 7.4] supplemented with 10% BSA and 1 ml/liter Proclin-300 as a preservative) was added across the entire plate. Plates were covered and gently shaken for 2 h at room temperature. Unbound conjugate was removed by washing. One hundred microliters of the single-component tetramethylbenzidine-peroxidase substrate (Bio-Rad) was added to all wells. After 30 min of incubation in the dark, the color reaction was stopped by the addition of 100 μl of diluted 0.36 N sulfuric acid in distilled water. Plates were read in an ELISA reader at 450 nm against a reference filter at 630 nm within 30 min after stopping.

Summary:
1. A random three-way ANOVA with day, operator, and sample as random factor was carried out (using proc mixed of SAS) on log10-transformed concentrations. Computations were done on transformed values, but CVs were expressed from nontransformed concentrations. The CV for repeatability of the assay was 4.72%. The global CV reproducibility and repeatability under routine conditions were 11.86%. Precision is in agreement with what is currently displayed by the ELISA. 2. In the reanalysis of seven studies in which different recombinant HBsAg-containing vaccines were used, a very good agreement in terms of seroprotection rate was observed after a full schedule and also 1 month postvaccination. Antibody GMCs were similar across the in-house and reference assays, with a trend toward overestimation using the in-house assay in the lower concentration range.

References:
1: Cambron P, Jacquet JM, Hoet B, Lievens M. Development and technical and clinical validation of a quantitative enzyme-linked immunosorbent assay for the detection of human antibodies to hepatitis B surface antigen in recipients of recombinant hepatitis B virus vaccine. Clin Vaccine Immunol. 2009 Aug;16(8):1236-46. Epub 2009 Jun 24. PubMed PMID: 19553553; PubMed Central PMCID: PMC2725549.