C4d High Sensitivity ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level High Sensitive Complement Component 4d (C4d) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level High Sensitive Complement Component 4d (C4d) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Basic complement C4; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 3; C4B_2; C4B1; C4B12; C4B2; C4B3; C4B5; C4BD; C4F; CH; Chido form of C4; CO4; complement C4-B; complement C4B1a; complement C4-B-like; complement component 4B (Chido blood group) ; CPAMD3.

Non-enzymatic component of the C3 and C5 convertases and thus essential for the propagation of the classical complement pathway. Covalently binds to immunoglobulins and immune complexes and enhances the solubilization of immune aggregates and the clearance of IC through CR1 on erythrocytes. C4A isotype is responsible for effective binding to form amide bonds with immune aggregates or protein antigens, while C4B isotype catalyzes the transacylation of the thioester carbonyl group to form ester bonds with carbohydrate antigens.

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 High Sensitive Complement Component 4d (C4d) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Sensitive Complement Component 4d (C4d) . 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 High Sensitive Complement Component 4d (C4d), 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 High Sensitive Complement Component 4d (C4d) 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) 96-103 99 EDTA Plasma (n=5) 93-101 98 Heparin Plasma (n=5) 78-88 82 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 96-103 99 Serum (n=5) 96-103 99 EDTA Plasma (n=5) 93-101 98 EDTA Plasma (n=5) 93-101 98 Heparin Plasma (n=5) 78-88 82 Heparin Plasma (n=5) 78-88 82

Component Amount Anti-C4d Microplate 96 Wells (12 x 8 Well Strips) C4d Lyophilized Standard 2 100X Biotinylated C4d 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-C4d Microplate 96 Wells (12 x 8 Well Strips) Anti-C4d Microplate 96 Wells (12 x 8 Well Strips) C4d Lyophilized Standard 2 C4d Lyophilized Standard 2 100X Biotinylated C4d Detector Antibody 120 uL 100X Biotinylated C4d 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 Complement Component 4d (C4d) . No significant cross-reactivity or interference between Complement Component 4d (C4d) and analogues was observed.