Human CXCL13/BLC/BCA-1 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY801
Ancillary Products Available
Human CXCL13 / BLC / BCA-1 ELISA Standard Curve
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Product Details
Procedure
Citations (7)
FAQs
Supplemental Products
Reviews (1)

Human CXCL13/BLC/BCA-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CXCL13/BCA-1. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

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; SCYB13B-cell-attracting chemokine 1; small inducible cytokine B subfamily (Cys-X-Cys motif), member 13 (B-cellchemoattractant)

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human CXCL13/BLC/BCA-1 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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  1. Activin A programs the differentiation of human TFH cells
    Nat Immunol, 2016;17(8):976-84.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. TGF-beta induces the differentiation of human CXCL13-producing CD4(+) T cells.
    Authors: Kobayashi S, Watanabe T, Suzuki R, Furu M, Ito H, Ito J, Matsuda S, Yoshitomi H
    Eur J Immunol, 2015;46(2):360-71.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Feasibility of the use of combinatorial chemokine arrays to study blood and CSF in multiple sclerosis.
    Authors: Edwards, Keith R, Goyal, Jaya, Plavina, Tatiana, Czerkowicz, Julie, Goelz, Susan, Ranger, Ann, Cadavid, Diego, Browning, Jeffrey
    PLoS ONE, 2013;8(11):e81007.
    Species: Human
    Sample Types: Serum
  4. Cerebrospinal fluid B cells correlate with early brain inflammation in multiple sclerosis.
    Authors: Kuenz B, Lutterotti A, Ehling R, Gneiss C, Haemmerle M, Rainer C, Deisenhammer F, Schocke M, Berger T, Reindl M
    PLoS ONE, 2008;3(7):e2559.
    Species: Human
    Sample Types: Serum
  5. Distinct cytokine-driven responses of activated blood gammadelta T cells: insights into unconventional T cell pleiotropy.
    Authors: Vermijlen D, Ellis P, Langford C, Klein A, Engel R, Willimann K, Jomaa H, Hayday AC, Eberl M
    J. Immunol., 2007;178(7):4304-14.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Distinct transcriptional programs activated by interleukin-10 with or without lipopolysaccharide in dendritic cells: induction of the B cell-activating chemokine, CXC chemokine ligand 13.
    Authors: Perrier P, Martinez FO, Locati M, Bianchi G, Nebuloni M, Vago G, Bazzoni F, Sozzani S, Allavena P, Mantovani A
    J. Immunol., 2004;172(11):7031-42.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Follicular dendritic cell regulation of CXCR4-mediated germinal center CD4 T cell migration.
    Authors: Estes JD, Thacker TC, Hampton DL, Kell SA, Keele BF, Palenske EA, Druey KM, Burton GF
    J. Immunol., 2004;173(10):6169-78.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

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Human CXCL13/BLC/BCA-1 DuoSet ELISA
By Anonymous on 11/29/2017
Sample Tested: Serum