| APO-54N-013-KI01 | 4x96 wells | 1,690.00 USD |  |
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Standard Curve Method: A 96-well ELISA plate was coated O/N at 4°C with 100µl/well of a 5µg/ml solution of MAb to BAFF (mouse) (5A8) (Prod. No. ALX-804-158) diluted in 50mM sodium bicarbonate buffer (pH 9.6). After two washes, wells were blocked with 100µl of PBS containing 1% BSA at 37°C for two hours. To reduce interference from rheumatoid factor (RF) in the serum, Ig were cleared by treating serum dilutions with 0.05 volume of protein G-Sepharose (Amersham Biosciences) twice for one hour at 4°C before pelleting the beads and collecting supernatants (precleared sera). A total of 20µl/well serum or rmsBAFF (Prod. No. ALX-522-052) serial dilutions in ELISA buffer (PBS, 0.05% Tween, 1% BSA) were added to the plates. After incubation for one hour at 37°C, wells were washed three times. Then, 100µl/well of a 2µg/ml solution of MAb to BAFF (mouse) (1C9) (Biotin) (Prod. No. ALX-804-157B) in PBS containing 1% BSA was added. After incubation for one hour at 37°C, wells were washed three times and incubated with 100µl/well of HRP-labeled streptavidin (DAKO) for 30 min. The plates were washed four times before the enzymatic reaction was conducted using the tetramethylbenzidine substrate kit (BD PharMingen). The reaction was stopped by adding 2M H2SO4, and the OD was measured at 450nm in an ELISA reader.Washing buffer: PBS containing 0.1% Tween 20.
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| Alternative Name: | BLyS, TALL-1, THANK, zTNF4, TNFSF 13B/20, CD257 |
| Sensitivity: | 0.3ng/ml (range 0 to 20ng/ml) |
| Assay Time: | 2.5-3 hours |
| Applications: | ELISA, Colorimetric detection |
| Application Notes: | For the quantitative determination of soluble mouse BAFF from biological fluids (serum and cell culture supernatant). |
| Wavelength: | 450 nm |
| Species reactivity: | Mouse |
| Contents: | 1 vial Standard (lyophilized) (STD) 1 vial Coating Antibody (COAT) 1 vial Detection Antibody (DET) |
| Handling: | For long term storage keep the protein standard at -20°C. Avoid freeze/thaw cycles. |
| Shipping: | Blue Ice |
| Long Term Storage: | -20°C |
| Scientific Background: | BAFF (B cell activation factor of the TNF family, also known as BLyS or TALL1) is a cytokine expressed predominantly by cells of the immune system such as neutrophils, monocytes, macrophages, dendritic cells, follicular dendritic cells, activated T cells and some malignant B cells. BAFF binds three distinct receptors (BAFF-R, TACI and BCMA) expressed predominantly on B cells, although activated T cells also express BAFF-R. BAFF is a master regulator of peripheral B cell survival, and also acts in processes such as immunoglobulin isotype switch and B cell co-stimulation. Besides its major role in B cell biology, BAFF co-stimulates activated T cells. Deregulated expression of this membrane-bound protein, which can readily be released in a soluble form, leads to autoimmune disorders in mice. In humans, elevated levels of soluble BAFF have been detected in the serum of patients with various autoimmune diseases. |
| UniProt ID: | Q9WU72 |
| Regulatory Status: | RUO - Research Use Only |
