HLA-B ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Major Histocompatibility Complex Class I B (MHCB) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Major Histocompatibility Complex Class I B (MHCB) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

AS; B-4901; HLA class I antigen HLA-B; HLA class I histocompatibility antigen, B alpha chain; HLAB; Human leukocyte antigen B; leukocyte antigen class I-B; major histocompatibility complex, class I, B; MHC class 1 antigen; MHC class I antigen B*7; MHC class I antigen HLA-B alpha chain; MHC class I antigen HLA-B heavy chain; MHC class I antigen SHCHA; MHC class I molecule; MHC HLA-B cell surface glycoprotein; MHC HLA-B transmembrane glycoprotein.

Antigen-presenting major histocompatibility complex class I (MHCI) molecule. In complex with B2M/beta 2 microglobulin displays primarily viral and tumor-derived peptides on antigen-presenting cells for recognition by alpha-beta T cell receptor (TCR) on HLA-B-restricted CD8-positive T cells, guiding antigen-specific T cell immune response to eliminate infected or transformed cells (PubMed:25808313, PubMed:29531227, PubMed:9620674, PubMed:23209413) . May also present self-peptides derived from the signal sequence of secreted or membrane proteins, although T cells specific for these peptides are usually inactivated to prevent autoreactivity (PubMed:7743181, PubMed:18991276) . Both the peptide and the MHC molecule are recognized by TCR, the peptide is responsible for the fine specificity of antigen recognition and MHC residues account for the MHC restriction of T cells (PubMed:29531227, PubMed:9620674, PubMed:24600035) . Typically presents intracellular peptide antigens of 8 to 13 amino acids that arise from cytosolic proteolysis via constitutive proteasome and IFNG-induced immunoproteasome (PubMed:23209413) . Can bind different peptides containing allele-specific binding motifs, which are mainly defined by anchor residues at position 2 and 9 (PubMed:25808313, PubMed:29531227) .|Involved in the presentation of foreign antigens to the immune system.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Major Histocompatibility Complex Class I B (MHCB) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Major Histocompatibility Complex Class I B (MHCB) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Major Histocompatibility Complex Class I B (MHCB), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Major Histocompatibility Complex Class I B (MHCB) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) serum (n=5) 95-104 99 EDTA plasma (n=5) 92-99 96 heparin plasma (n=5) 82-96 91 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 95-104 99 serum (n=5) 95-104 99 EDTA plasma (n=5) 92-99 96 EDTA plasma (n=5) 92-99 96 heparin plasma (n=5) 82-96 91 heparin plasma (n=5) 82-96 91

Component Amount Anti-HLA-B Microplate 96 Wells (12 x 8 Well Strips) HLA-B Lyophilized Standard 2 x 10 ng 100X Biotinylated HLA-B Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-HLA-B Microplate 96 Wells (12 x 8 Well Strips) Anti-HLA-B Microplate 96 Wells (12 x 8 Well Strips) HLA-B Lyophilized Standard 2 x 10 ng HLA-B Lyophilized Standard 2 x 10 ng 100X Biotinylated HLA-B Detector Antibody 120 uL 100X Biotinylated HLA-B Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Major Histocompatibility Complex Class I B (MHCB) . No significant cross-reactivity or interference between Major Histocompatibility Complex Class I B (MHCB) and analogues was observed.