Human CXCL13/BLC/BCA-1 Antibody

Catalog #: MAB8012 Datasheet / COA / SDS

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MAB8012-100
MAB8012-SP

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Human CXCL13/BLC/BCA-1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human CXCL13/BLC/BCA-1 in direct ELISAs.

Source

Monoclonal Mouse IgG₁ Clone # 53602

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human CXCL13/BLC/BCA-1
Val23-Arg94
Accession # O43927

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Immunohistochemistry

1-25 µg/mL

See below

Intracellular Staining by Flow Cytometry

10 µg/10⁶ cells

Immature Dendritic cells were used (CD14 selected PBMCs are treated with 20 ng/mL rhIL-4 (Catalog # 204-IL) and 50 ng/mL rhGM-CSF (Catalog # 215-GM) for 6 days)

Neutralization

Measured by its ability to neutralize CXCL13/BLC/BCA‑1-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CXCR5. The Neutralization Dose (ND₅₀) is typically 0.5-2.5 µg/mL in the presence of 0.05 µg/mL Recombinant Human CXCL13/BLC/BCA‑1.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry

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CXCL13/BLC/BCA-1 in Human Tonsil. CXCL13/BLC/BCA-1 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human CXCL13/BLC/BCA-1 Monoclonal Antibody (Catalog # MAB8012) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Neutralization

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Chemotaxis Induced by CXCL13/BLC/BCA-1 and Neutralization by Human CXCL13/BLC/BCA-1 Antibody. Recombinant Human CXCL13/BLC/BCA-1 (801-CX) chemoattracts the BaF3 mouse pro-B cell line transfected with human CXCR5 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (AR002). Chemotaxis elicited by Recombinant Human CXCL13/BLC/BCA-1 (0.05 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CXCL13/BLC/BCA-1 Monoclonal Antibody (Catalog # MAB8012). The ND₅₀ is typically 0.5-2.5 µg/mL.

Intracellular Staining by Flow Cytometry

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Detection of CXCL13/BLC/BCA‑1 in Immature dendritic cells by Flow Cytometry. Immature dendritic cells (CD14 selected PBMCs are treated with 20 ng/mL Recombinant Human IL-4 Protein (204-IL) and 50 ng/mL Recombinant Human GM-CSF Protein (215-GM) for 6 days) were stained with Mouse Anti-Human CXCL13/BLC/BCA‑1 Monoclonal Antibody (Catalog # MAB8012, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (FC012). View our protocol for Staining Intracellular Molecules.

Intracellular Staining by Flow Cytometry

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Detection of CXCL13/BLC/BCA‑1 in Immature Dendritic cells by Flow Cytometry. Immature dendritic cells (CD14+ selected PBMCs are treated with 20 ng/mL Recombinant Human IL-4 Protein (204-IL) and 50 ng/mL Recombinant Human GM-CSF Protein (215-GM) for 6 days) were stained with Mouse Anti-Human CD11c PE‑conjugated Monoclonal Antibody (Catalog # FAB1777P) and either (A) Mouse Anti-Human CXCL13/BLC/BCA‑1 Monoclonal Antibody (Catalog # MAB8012) or (B) Mouse IgG₁ Isotype Control (Catalog # MAB002) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CXCL13/BLC/BCA-1

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.

References

Gunn, M.D. et al. (1998) Nature, 391:799.
Legler, D.F. et al. (1998) J. Exp. Med. 187:655.
Forster, R. et al. (1996) Cell 87:1037.

Entrez Gene IDs

10563 (Human); 55985 (Mouse)

Alternate Names

ANGIE; ANGIE2; B cell-attracting chemokine 1; B lymphocyte chemoattractant; BCA1; BCA-1; BCA1B-cell chemoattractant; BCA-1CXC chemokine BLC; B-cell-homing chemokine (ligand for Burkitt's lymphoma receptor-1); BLC; BLCSmall-inducible cytokine B13; BLR1L; B-lymphocyte chemoattractant; chemokine (C-X-C motif) ligand 13 (B-cell chemoattractant); chemokine (C-X-C motif) ligand 13; C-X-C motif chemokine 13; CXCL13; SCYB13; SCYB13B-cell-attracting chemokine 1; small inducible cytokine B subfamily (Cys-X-Cys motif), member 13 (B-cellchemoattractant)