Cotton Rat CXCL1/2/3/GRO Antibody

  
  • Species Reactivity
    Cotton Rat
  • Specificity
    Detects cotton rat CXCL1/2/3/GRO in direct ELISAs and Western blots. In direct ELISAs, this antibody shows 5-20% cross-reactivity with recombinant rat CXCL1 and recombinant human CXCL2 and no cross-reactivity with recombinant cotton rat CXCL10, recombinant human CXCL1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12/SDF-1 alpha, rh12/SDF-1 beta, 13, 16, recombinant mouse CXCL1, 2, 4, 6, 7, 9, 10, 11, 12/SDF-1 alpha, 13, 15, 16, 17, recombinant rat (rr) CXCL2, rrCXCL3/CINC-2 alpha, rr3/CINC-2 beta, or recombinant porcine CXCL8.
  • Source
    Monoclonal Mouse IgG1 Clone # 186413
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant cotton rat GRO
    Ala28-Lys100
    Accession # AAL16934
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied 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
  • Western Blot
    1 µg/mL
    Recombinant Cotton Rat CXCL1/2/3/GRO (Catalog # 1114-GR)
  • Neutralization
    Measured by its ability to neutralize CXCL1/2/3/GRO-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CXCR2. The Neutralization Dose (ND50) is typically 0.08-0.4 µg/mL in the presence of 5 ng/mL Recombinant Cotton Rat CXCL1/2/3/GRO.
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.
Data Examples
Chemotaxis Induced by CXCL1/2/3/GRO and Neutralization by Cotton Rat CXCL1/2/3/GRO Antibody. Recombinant Cotton Rat CXCL1/2/3/GRO (Catalog # 1114-GR) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CXCR2 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Cotton Rat CXCL1/2/3/GRO (5 ng/mL) is neutralized (green line) by increasing concentrations of Cotton Rat CXCL1/2/3/GRO Monoclonal Antibody (Catalog # MAB1114). The ND50 is typically 0.08-0.4 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile 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: CXCL1/2/3/GRO

Cotton rat growth-regulated protein (GRO) is a member of the ELR+ CXC subfamily of chemokines (1). The cotton rat GRO cDNA encodes a 100 amino acid (aa) precursor protein with a 27 aa putative signal peptide and a 73 aa mature protein (2). Among the three human GRO proteins, mature cotton rat GRO is most closely related to CXCL1/GRO alpha, sharing approximately 77% aa sequence identity (3). It also shares 75% and 70% aa sequence identity with mature human CXCL2/GRO beta and CXCL3/GRO gamma, respectively (4). Compared with rat CXC chemokines, mature cotton rat shows 92%, 76%, and 76% aa sequence homology with rat KC/GRO/CINC-1, CINC-2, and CINC-3, respectively (5). Mature cotton rat CXCL1 also shares 74% sequence homology with mature cotton rat MIP-2 (6). Similarly to GRO proteins from other species, cotton rat CXCL1 is expected to be a potent neutrophil chemoattractant and activator. Cotton rat CXCL1 has been shown to bind and activate both mouse chemokine receptors CXCR1 and CXCR2 (7). Similarly to human CXCL1, the cotton rat CXCL1 concentration required to activate CXCR1 is at least an order of magnitude higher than that required to activate CXCR2 (7, 8). Based on studies on rodents, cotton rat CXCL1 is predicted to be produced by a wide variety of cell types including macrophages, endothelial cells, fibroblasts, astrocytes, neutrophils, keratinocytes, and intestinal epithelium (1, 9).

  • References:
    1. Rollins, B.J. (1997) Blood 90:909.
    2. Blanco, J.C. et al. (2001) GenBank Accession #: AAL16934.
    3. Richmond, A. et al. (1988) EMBO J. 7:2025.
    4. Haskill, S. et al. (1990) Proc. Natl. Acad. Sci. USA 87:7732.
    5. Nakagawa, H. et al. (1994) Biochem. J. 301:545.
    6. Blanco, J.C. et al. (2001) GenBank Accession #: AAL26705.
    7. R&D Systems, unpublished observations.
    8. Schumacher, C. et al. (1992) Proc. Natl. Acad. Sci. USA 89:10542.
    9. Zlotnik, A.O. (2000) Immunity 12:121.
  • Alternate Names:
    CXCL1/2/3
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