Mouse CXCL13/BLC/BCA-1 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Mouse BLC/BCA-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse BLC/BCA-1 in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse BLC/BCA-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse BLC/BCA-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse BLC/BCA-1.

Preparation and Storage

Background: CXCL13/BLC/BCA-1

CXCL13/BLC/BCA-1 is a constitutively expressed chemokine that plays an important role in B and T cell homing. It is expressed by salivary gland epithelium, dendritic cells, osteoclasts, and peritoneal macrophages. It can form homodimers or heterodimers with FGF basic, and it signals through CXCR3 or CXCR5. CXCL13 induces the migration of naïve B cells and a subset of memory T cells to lymphoid tissue. It also promotes B1 cell migration into the omentum and peritoneum. In the fetus, CXCL13 attracts CD4+ CD3- IL-7 R alpha+ hematopoietic cells to sites of future Peyer’s patch development.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.