4VS4
F501, F529, F550, F571
F173, F227, F231, F240, SAA19cc, SAA21cc, VSA31cc, VSA34cc, VSA38cc
VSA2, VSA43
Recombinant chimeric antibody expressed in a mammalian cell line, composed of original wild type variable domains of rat derived MAb and human IgG1 constant domains (F501, F529, F550, F571)
Hybridoma clones have been derived from hybridization of Sp2/0 myeloma cells with spleen cells of Balb/c mice immunized with either:
feline SAA (F173, F227, F231, F240), or
human SAA (SAA19cc, SAA21cc), or
canine SAA (VSA31cc, VSA34cc, VSA38cc), or
synthetic peptides derived from the region 79-104 a.a.r. of canine SAA (VSA2, VSA43)
IgG1 for F501, F529, F550, F571, F227, F231, VSA2
IgG2a for F173, F240, SAA19cc, VSA31cc, VSA38cc
IgG2b for SAA21cc, VSA34cc, VSA43
Protein A chromatography
PBS, pH 7.4, 0.09 % sodium azide (NaN3), 5 mM EDTA for F501, F529, F550, F571
PBS, pH 7.4, 0.09 % sodium azide (NaN3)for F173, F227, F231, F240, SAA19cc, VSA31cc, VSA34cc, VSA38cc, VSA2, VSA43
50 mM citrate, 150 mM NaCl, pH 6.0, 0,09% azide (NaN3) for SAA21cc
+4 °C (+2 … +8 °C allowed)
Pair SAA19cc – VSA34cc:
Plates blocking with casein is needed to prevent non-specific binding of SAA to the wells of an immunoassay plate. We recommend avoiding using bovine serum albumin (BSA) as a buffer component or blocking agent. In buffers, BSA can be replaced with 1% casein. According to our data, Tween 20 lowered the signal when it was used in concentration 0.05-0.1% in antigen dilution buffer. When lower concentration of Tween 20 was used (0.005-0.025%), non-specific binding of SAA to the wells of an immunoassay plate was observed. Therefore, we recommend starting assay optimization using 0.01% CHAPS in antigen dilution and washing buffers. We recommend carrying out immunoassay at room temperature.
Pairs VSA2 – VSA38cc, VSA2 – VSA31cc, VSA38cc – VSA43, F571 – F173: Plates blocking with casein is needed to prevent non-specific binding of SAA to the wells of an immunoassay plate. For pairs VSA2 – VSA38cc, VSA2 – VSA31cc and VSA38cc – VSA43 we recommend avoiding using BSA as a buffer component or blocking agent. In buffers, BSA can be replaced with 1% casein. We recommend using antigen dilution buffer containing 0.05% Tween 20. We recommend carrying out immunoassay at 37 °C.
Pairs F227 – F529, F231 – F550, F240 – F501, F240 – F550: Plate blocking is not needed. We recommend using antigen dilution buffer containing 10 mg/ml BSA and 0.05% Tween 20. We recommend carrying out immunoassay at room temperature.
This product is sold for research or further manufacturing use only. Standard Laboratory Practices should be followed when handling this material.
Product contains sodium azide as a preservative. Although the amount of sodium azide is very small appropriate care must be taken when handling this product.
(Catalogue number and MAb: 4VS4-)