Anti-Human BCA-1

Product Datasheet

www.leinco.com

excellence in early discovery research

Anti-Human B Cell-Attracting Chemokine-1 (BCA-1)

Purified No Carrier Protein

(CXCL13, BLC (mouse), BLR1 Ligand, SCYB13, ANGIE, ANGIE2)

Monoclonal Antibody

Prod. No.:	C1457
Clone:	53610
Isotype:	Mouse IgG₁
Pkg. Size:	1 mg, 500 µg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Mouse Anti-Human B Cell-Attracting Chemokine-1 (BCA-1) (Clone 53610) recognizes an epitope on Human BCA-1. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:

CXCL13, also known as B-lymphocyte chemoattractant (BLC), is a CXC chemokine that is constitutively expressed in secondary lymphoid organs. BCA-1 cDNA encodes a protein of 109 amino acid residues with a leader sequence of 22 residues. Mature human BCA-1 shares 64% amino acid sequence similarity with the mouse protein and 23 - 34% amino acid sequence identity with other known CXC chemokines. Recombinant or chemically synthesized BCA-1 is a potent chemoattractant for B lymphocytes but not T lymphocytes, monocytes or neutrophils. BLR1, a G protein-coupled receptor originally isolated from Burkitt’s lymphoma cells, has now been shown to be the specific receptor for BCA-1. Among cells of the hematopoietic lineages, the expression of BLR1, now designated CXCR5, is restricted to B lymphocytes and a subpopulation of T helper memory cells. Mice lacking BLR1 have been shown to lack inguinal lymph nodes. These mice were also found to have impaired development of Peyer’s patches and defective formation of primary follicles and germinal centers in the spleen as a result of the inability of B lymphocytes to migrate into B cell areas.

Known Species Cross Reactivity: Hu+

Host Species

Mouse

Immunogen

Purified Recombinant Human BCA-1 (>98%)

Formulation

This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.

Reconstitution

This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1.0 ml. This will result in a final antibody concentration of 500 µg/ml.

Endotoxin

<0.1 EU/µg as determined by the LAL method

Storage and Stability

The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.

Reported Applications

ELISA Cap, FC, N, WB

ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 2-8 µg/ml. A suitable detection antibody is Anti-Human BCA-1-Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: B116) at a concentration of approximately 0.1-0.4 µg/ml. A suggested standard for this assay is Recombinant Human BCA-1 (Leinco Prod. No.: B111).

Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing BCA-1. A suggested method would be to stain cells expressing BCA-1 with 0.25 µg per 1 - 2.5 x 10⁵ cells in a total staining volume of ≤200 µl followed by Goat Anti-Mouse IgG (H&L)-R-phycoerythrin (Leinco Prod. No.: M1189).

Neutralization: This antibody is useful for neutralization of Human BCA-1 bioactivity. The antibody dose required to neutralize 50% (ND₅₀) of the biological activity of Human BCA-1 (at 0.05 ng/ml) is 0.3 - 0.9 µg/ml.

Western Blotting: To detect Human BCA-1 this monoclonal antibody can be used at a concentration of 0.1 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human BCA-1 (Leinco Prod. No.: B111).

Each investigator should determine their own optimal working dilution for specific applications.

Country of Origin

USA