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|Sample type||Average %||Range|
|Serum||95.1||92.4% - 97.7%|
|Cell culture media||103.7||98.9% - 107.5%|
|Heparin Plasma||106||103.8% - 107.9%|
|EDTA Plasma||101.7||99.2% - 107%|
|Citrate Plasma||101.2||96.7% - 111.3%|
Abcam’s BCA1 (CXCL13) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of BCA1 protein in plasma, serum and culture supernatants.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Human B-lymphocyte chemoattractant/B Cell-attracting Chemokine 1 (BCA1/CXCL13) is a member of the CXC chemokine superfamily. BCA1 (CXCL13) is produced by stromal cells from the liver, spleen, lymph nodes and the gut. It binds to the CXCR5 receptor in lymphoid follicles controlling the homing signal of B cells and subsets of T cells to the lymphoid nodes, compartmentalizing lymphocytes and antigen presenting cells within the follicles of lymphoid tissues. BCA-1 (CXCL13) is also required for the embryonic development of most of lymph nodes and Peyer’s patches. CXCL13-/- as well as CXCR5-/- mice lack secondary lymphoid structures and show a disrupted splenic architecture.
Furthermore, BCA1 (CXCL13) is considered an essential chemokine critical for the formation of ectopic lymphoid tissues within the central nervous system (CNS) and for the chemoattraction of B cells to the CNS in multiple neurological diseases. Elevated levels of the protein have been found in the cerebrospinal fluid of patients with Multiple Sclerosis, Neuroborreliosis, Neurosyphilis and Neuromyelitis optica. Elevated levels of BCA-1(CXCL13) in serum have also been correlated with HIV non Hodgking B cell lymphoma, Post-transplant lymphoproliferative disease and childhood-onset Lupus disease.
|Components||1 x 96 tests|
|10X BCA-1(CXCL13) Capture Antibody||1 x 600µl|
|10X BCA-1(CXCL13) Detector Antibody||1 x 600µl|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|Antibody Diluent CPI - HAMA Blocker (ab193969)||1 x 6ml|
|BCA-1(CXCL13) Human Lyophilized Recombinant/Purified Protein||2 vials|
|Plate Seals||1 unit|
|Sample Diluent NS (ab193972)||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Development Solution||1 x 12ml|
Our Abpromise guarantee covers the use of ab179881 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Background-subtracted data values (mean +/- SD) are graphed.
THP-1 cells were cultured in RPMI supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin. Cells were cultured for 8 hours with 1 µg/mL of Human IFNγ recombinant protein followed with 1 µg/mL of LPS or vehicle overnight at 37⁰C. The concentrations of BCA1 (CXCL13) were interpolated from the calibration curve and corrected for sample dilution. The mean BCA1 (CXCL13) concentration was determined to be 8.4 pg/mL in IFNγ only treated cells and 241 pg/mL in IFNγ/LPS treated cells.
Human PBMC were cultured in RPMI supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin. Cells were cultured for 2 days at 37⁰C in the presence or absence of PHA. The concentrations of BCA1 (CXCL13) were interpolated from the calibration curve and corrected for sample dilution. The mean BCA1 (CXCL13) concentration was determined to be undetectable in unstimulated PBMC supernatants and 22.3 pg/mL in stimulated PBMCs
ab179881 has not yet been referenced specifically in any publications.
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