Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, ELISA Capture (Matched Antibody Pair)

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 131120
View all CXCL14/BRAK Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CXCL14/BRAK
Leu23-Tyr99
Accession # O95715

Specificity

Detects human CXCL14/BRAK in ELISAs and Western blots. In direct ELISAs, shows approximately 80% cross-reactivity with recombinant mouse (rm) CXCL14/BRAK and no cross-reactivity with recombinant human (rh)GRO alpha, rhGRO beta, rhGRO gamma,rhMIP-1 alpha, rmMIP-1 alpha, rmMIP-1 beta, rhMIP-1 delta, rmMIP-1 gamma, rmMIP-2, rhMIP-3 alpha, rmMIP-3 alpha, rrMIP-3 alpha, rhMIP-3 beta, or rmMIP-3 beta.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human CXCL14/BRAK Antibody (131120)

Human CXCL14 / BRAK ELISA Standard Curve

Human CXCL14 / BRAK ELISA Standard Curve

Recombinant Human CXCL14/BRAK (Catalog # 866-CX) was serially diluted and captured by Mouse Anti-Human CXCL14/BRAK Monoclonal Antibody (Catalog # MAB866) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human CXCL14/BRAK Biotinylated Monoclonal Antibody (Catalog # ) was incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Detection of CXCL14/BRAK by Western Blot

Detection of CXCL14/BRAK by Western Blot

Glioblastoma cells stably overexpressing CXCL14 show enhanced proliferation and migration. (A) qRT-PCR analysis of CXCL14 mRNA expression in U87MG (upper panel) or LN229 (lower panel) cells transfected with either the empty vector pcDNA3.1/V5-HisC (pcDNA), or the CXCL14-expressing vector pCXCL14-V5. In both panels, data are expressed as compared to empty vector-transfected cells, where CXCL14 expression was set as = 1. (B) Western blot analysis showing CXCL14 protein expression in U87MG (upper panel) or LN229 (lower panel) cells transfected with either the empty vector pcDNA3.1/V5-His C, or the CXCL14-expressing vector pCXCL14-V5. In the immunoblots shown on the left, ectopic CXCL14 was revealed by using an anti-V5 antibody, while on the right a CXCL14-specific antibody was used. beta -actin detection was used as a loading control. (C) Proliferation (MTS assay) of U87MG (left) or LN229 (right) cells stably transfected with either the empty vector pcDNA3.1/V5-His C, or the CXCL14-expressing vector pCXCL14-V5. Results are shown as the mean ± S.D. and represent the average of two experiments performed in triplicate. Data were analyzed by a two-tailed unpaired Student’s t-test. * p < 0.05 ** p < 0.01. (D) Boyden chamber migration assay performed with U87MG cells stably transfected with either the empty vector pcDNA3.1/V5-HisC (pcDNA), or the CXCL14-expressing vector pCXCL14-V5. The graph depicts the relative migration of CXCL14-expressing cells compared to empty vector transfected ones, whose migration ability was set as = 1. Results are presented as mean ± S.D. with significant differences from controls (*) shown (p < 0.05). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31117166), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human CXCL14/BRAK Antibody (131120)

Application
Recommended Usage

Western Blot

1 µg/mL
Sample: Recombinant Human CXCL14/BRAK (Catalog # 866-CX)

Human CXCL14/BRAK Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human CXCL14/BRAK Biotinylated Antibody (Catalog # BAM8661)
  • Standard: Recombinant Human CXCL14/BRAK Protein, CF (Catalog # 866-CX)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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. *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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CXCL14/BRAK

CXCL14/BRAK (breast and kidney-expressed chemokine), also named MIP-2 gamma, KEC (kidney-expressed chemokine), and BMAC (B cell and monocyte-activating chemokine), is a member of CXC chemokine superfamily (1 - 5). The deduced 99 amino acid (aa) residue precursor has a 22 aa putative signal peptide that is cleaved to produce the 77 aa mature protein. Mature human and mouse CXCL14 differ by only 2 residues. Human CXCL14 shares approximately 30% aa sequence identity with MIP-2 alpha (GRO beta ) as well as MIP-2 beta  (GRO gamma ). The gene for CXCL14 has been mapped human chromosome 5q31. Unlike the MIP-2 chemokines, CXCL14 lacks the ELR domain preceding the CXC motif. CXCL14 transcripts are constitutively expressed at high levels in the basal layer of epidermal keratinocytes and dermal fibroblasts of skin tissues as well as lamina propria cells in normal intestinal tissues. CXCL14 has been shown to be a highly selective chemoattractant for monocytes that have been treated with prostaglandin E2 or forskolin, agents that activate adenylate cyclase. CXCL14 has been proposed to be important for regulating the trafficking of macrophage precursor to regions in skin and mucosal tissues that support their development. Consistent with this hypothesis, macrophages were frequently found to co-localize with CXCL14-producing cells in the dermis and lamina propria.

References

  1. Hromas, R. et al. (1999) Biochem. Biophys. Res. Commun. 255:703.
  2. Cao, X. et al. (2000) J. Immunol. 165:2588.
  3. Kurth, I. et al. (2001) J. Exp., Med. 194:855.
  4. Frederick, M.J. et al. (2000) Am. J. Pathol. 156:1937.
  5. Sleeman, M.A. et al. (2000) Int. Immunol. 12:677.

Alternate Names

Bolekine, BRAK, CXC-X3

Entrez Gene IDs

9547 (Human); 57266 (Mouse)

Gene Symbol

CXCL14

UniProt

O95715

Additional CXCL14/BRAK Products

Product Documents for Human CXCL14/BRAK Antibody (131120)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CXCL14/BRAK Antibody (131120)

For research use only

Related Research Areas

Chemokines and Receptors

Citations for Human CXCL14/BRAK Antibody (131120)

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