Recombinant Human CXCL13/BLC/BCA-1 (Catalog # 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. Chemotaxis elicited by Recombinant Human CXCL13/BLC/BCA-1 (50 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CXCL13/BLC/BCA-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF801). The ND₅₀ is typically 1-4 µg/mL.

CXCL13/BLC/BCA‑1 in Human Lymphoma.

CXCL13/BLC/BCA-1 was detected in immersion fixed paraffin-embedded sections of human lymphoma using Goat Anti-Human CXCL13/BLC/BCA-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF801) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of CXCL13/BLC/BCA-1 in Human Lymphoma.

Formalin-fixed paraffin-embedded tissue sections of Hodgkin’s Lymphoma were probed for CXCL13 mRNA (ACD RNAScope Probe, catalog # 311321; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using goat anti-human CXCL13 polyclonal antibody (R&D Systems catalog # AF801) at 5ug/mL with 1-hour incubation at room temperature followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm.

Detection of CXCL13/BLC/BCA-1 by Flow Cytometry

The anti-tumor function of Tfh cells was impaired in PDAC. (A) Representative flow cytometry data and the proportion of functional cTfh cells (CXCL13+ IL-21+) from the peripheral blood of healthy donors (n = 4) and patients with PDAC of different stages (nI = 5, nII = 4, nIII = 4). (B) Representative flow cytometry data and (C) the proportion of CD8+ T cells and B cells recruited to the bottom of the Transwell chamber by cTfh cells treated with or without rhCXCL13 or anti-CXCL13 neutralizing antibodies. (D) Representative flow cytometry data and the proportion of plasma cells (CD27+ CD38+) in B cells (CD19+ CD20+) from the peripheral blood of healthy donors (n = 4) and patients with PDAC at different stages (nI = 5, nII = 4, nIII = 4). (E) Representative flow cytometry data and the proportion of IgG-secreting plasma cells from cells in (D). (F) Representative flow cytometry data and the proportion of IgM-secreting plasma cells from cells in (D). (G) Representative flow cytometry data and the proportion of plasma cells (CD27+ CD38+) in B cells (CD19+ CD20+) co-cultured with cTfh cells sorted from the peripheral blood of healthy donors (n = 3) and patients with PDAC (n = 3). (H) Representative flow cytometry data and the proportion of IgG-secreting plasma cells from cells in (G). (I) Representative flow cytometry data and the proportion of IgM-secreting plasma cells from cells in (G). Asterisks indicated the significance level of the p-value (* p < 0.05 and ** p < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34359579), licensed under a CC-BY license. Not internally tested by R&D Systems.

Human CXCL13 / BLC / BCA-1 ELISA Standard Curve

Recombinant Human CXCL13/BLC/BCA-1 (Catalog # 801-CX) was serially diluted and captured by Mouse Anti-Human CXCL13/BLC/BCA-1 Monoclonal Antibody (Catalog # MAB801) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human CXCL13/BLC/BCA-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF801) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

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Applications for Human CXCL13/BLC/BCA-1 Antibody

Application

Recommended Usage

Dual RNAscope ISH-IHC Compatible

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymphoma

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil and human lymphoma

Western Blot

0.1 µg/mL
Sample: Recombinant Human CXCL13/BLC/BCA-1 (Catalog # 801-CX)

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 1-4 µg/mL in the presence of 50 ng/mL Recombinant Human CXCL13/BLC/BCA-1.

Reviewed Applications

Read 7 reviews rated 4.6 using AF801 in the following applications:

Flow Cytometry (1 Review)
Immunocytochemistry/Immunofluorescence (1 Review)
Immunofluorescence - paraffin (1 Review)
Immunohistochemistry (3 Reviews)
Immunohistochemistry-Paraffin (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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 BCA1. 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.

Alternate Names

ANGIE2, BCA-1, BCA1, BLC, BLR1L, SCYB13

Entrez Gene IDs

10563 (Human); 55985 (Mouse)

Gene Symbol

CXCL13

UniProt

Q53X90

Additional CXCL13/BLC/BCA-1 Products

Product Documents for Human CXCL13/BLC/BCA-1 Antibody

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Product Specific Notices for Human CXCL13/BLC/BCA-1 Antibody

For research use only

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