Detects human CXCL1/GRO alpha /KC/CINC-1 in direct ELISAs and Western blots. In direct ELISAs, less than 50% cross-reactivity with recombinant human (rh) GRO beta and rhGRO gamma is observed, less than 25% cross-reactivity with recombinant rat (rr) CINC-1 is observed and less than 15% cross-reactivity with recombinant mouse (rm) KC is observed. In Western blots, no cross-reactivity with rhNAP-2 or rhPF-4 is observed.
Polyclonal Goat IgG
E. coli-derived recombinant human CXCL1/GRO alpha /KC/CINC-1 (R&D Systems, Catalog # 275-GR) Ala35-Asn107 Accession # P09341
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.
<0.10 EU per 1 μg of the antibody by the LAL method.
Measured by its ability to neutralize CXCL1/GRO alpha /KC/CINC‑1-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CXCR2. The Neutralization Dose (ND50) is typically 0.03-0.15 µg/mL in the presence of 0.01 µg/mL Recombinant Human CXCL1/GRO alpha /KC/CINC‑1.
Please Note: Optimal dilutions should be determined by each laboratory for each application.
are available in the Technical Information section on our website.
Detection of Recombinant Human CXCL1/GRO alpha /KC/CINC‑1 by Western Blot.
Western blot shows 25 ng of Recombinant Human CXCL1/GRO alpha /KC/CINC‑1 (Catalog # 275-GR), Recombinant Mouse CXCL1/GRO alpha /KC/CINC‑1 aa 20-96 (Catalog # 453-KC), Recombinant Rat CXCL1/GRO alpha /KC/CINC‑1 (Catalog # 515-CN), Recombinant Human CXCL2/GRO beta /MIP‑2/CINC‑3 aa 35-107 (Catalog # 276-GB), Recombinant Human CXCL3/GRO gamma /CINC‑2/DCIP‑1 (Catalog # 277-GG), Recombinant Human CXCL7/NAP‑2 (Catalog # 393-NP), and Recombinant Human CXCL4/PF4 (Catalog # 795-P4). PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Human CXCL1/GRO alpha /KC/CINC‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF275) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for CXCL1/GRO alpha /KC/CINC‑1 at approximately 11 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Chemotaxis Induced by CXCL1/GRO alpha and Neutralization by Human CXCL1/GRO alpha Antibody.
Recombinant Human CXCL1/GRO alpha (Catalog # 275-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 Human CXCL1/GRO alpha (0.01 µg/mL) is neutralized (green line) by increasing concentrations of Human CXCL1/GRO alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF275). The ND50 is typically 0.03-0.15 µg/mL.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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/GRO alpha/KC/CINC-1
The gene for CXCL1/GRO alpha was initially discovered in hamster cells, using subtractive hybridization techniques, as a message that is over-expressed in tumorigenic cells and in normal cells during growth stimulation. The hamster cDNA was cloned and used as a probe for the subsequent cloning of the human GRO cDNA. Independently, a cDNA encoding a secreted protein with melanoma growth stimulating activity (MGSA) was also cloned from a human melanoma cell line and found to be identical to GRO. In addition to the initially cloned GRO gene, now designated CXCL1, two additional GRO genes, GRO beta or MIP-2 alpha and GRO gamma or MIP‑2 beta, which shared 90% and 86% amino acid sequence homology, respectively, with CXCL1, have been identified. All three human GROs are members of the alpha (C-X-C) subfamily of chemokines. The three GRO cDNAs encode 107 amino acid precursor proteins from which the N-terminal 34 amino acid residues are cleaved to generate the mature GROs. There are no potential N-linked glycosylation sites in the amino acid sequences. GRO expression is inducible by serum or PDGF and/or by a variety of inflammatory mediators, such as IL-1 and TNF, in monocytes, fibroblasts, melanocytes, and epithelial cells. In certain tumor cell lines, GRO is expressed constitutively. Similar to other alpha chemokines, the three GRO proteins are potent neutrophil attractants and activators. In addition, these chemokines are also active toward basophils. All three GROs can bind with high affinity to the IL-8 receptor type B. The rat homolog of human CXCL1, CINC, is much more active than human CXCL1 on rat neutrophils, suggesting that this cytokine may have selective species specificity.
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