CXCL10 Antibody (OASA07911)

Catalog#: OASA07911

• Size: 50ug

• Description of Target: RABBIT ANTI HUMAN IP-10:Biotin
• Gene Symbol: CXCL10
• Alias Symbols: C7; IFI10; INP10; IP-10; crg-2; mob-1; SCYB10; gIP-10; CXCL10
• Protein Accession#: NP_001556.2
• Swissprot Id: P02778
• Host: Rabbit
• Protein Size (# AA): 98
• Specificity: IP-10
• Application: ELISA, WB
• Conjugation: Biotin
• Immunogen: The immunogen for anti-CXCL10 antibody: recombinant human IP-10
• Product Format: Purified IgG conjugated to Biotin - lyophilised
• Isotype: Polyclonal IgG
• Predicted Homology Based on Immunogen Sequence: Human
• Applications Info: ELISA:    0.15 - 0.30ug/ml
  Western Blotting:    0.1 - 0.2ug/ml
  
  Preservative Stabilisers: None present.
  
  Antiserum Preparation: Antisera to human IP-10 were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG prepared by affinity chromatography.
  Approx Protein Conc: IgG concentration 50ug/ml
  
  Buffer Solutions: Phosphate buffered saline
• Key Reference: 1. Cox, M.A. et al. (2001) Human interferon-inducible 10-kDa protein and human interferon-inducible T cell alpha chemoattractant are allotopic ligands for human CXCR3: Differential binding to receptor states. Mol. Pharmacol. 59: 707-715.
  2. Sauty, A. et al. (1999) The T cell-specific CXC chemokines IP-10, Mig and I-TAC are expressed by activated human bronchial epithelial cells. J. Immunol. 162: 3549-3558.
• Reconstitution and Storage: Reconstitution: Reconstitute with 1.0ml sterile PBS containing 0.1% Bovine Serum Albumin.
  Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Aviva Systems Biology recommend that the vial is gently

Product Protocol: CXCL10 antibody used to evaluate expression of the th1 chemokine ifn-gamma-inducible protein 10 in the airway alters mucosal allergic sensitization in mice (OASA07911)

Specificity: IP-10
Product Page: CXCL10 antibody (OASA07911)
Antibody Pair Available: OASA07910 Capture OASA07911 Detection
Experiment Type: Cytokine and Ig measurement

Protocol:
ELISA kits for IL-4, IFN-γ, eotaxin, monocyte chemoattractant protein (MCP) 1, MIP-1α, and RANTES were purchased from R&D Systems (Minneapolis, MN), while the kit for IL-5 was obtained from Amersham (Buckinghamshire, U.K.). IP-10 was measured by ELISA according to a standard alkaline phosphatase/streptavidin assay protocol (R&D Systems). Briefly, IP-10 was captured with an anti-human IP-10 Ab in the solid phase and developed with biotinylated anti-human IP-10 (R&D Systems); recombinant human IP-10 (PeproTech, Rocky Hill, NJ) was used as a standard. Levels of OVA-specific IgE were detected using an Ag-capture (biotinylated OVA) ELISA method; the ELISA was standardized with serum obtained from mice sensitized to OVA according to a conventional i.p. sensitization model and bled 24 h after the second sensitization. OVA-specific IgG2a was measured by sandwich ELISA with OVA in the solid phase. Ninety-six-well plates were coated with 5 μg/ml OVA in borate buffer (100 μl/well) for 1 h at 37°C, 3 h at room temperature, and then overnight at 4°C. Plates were blocked for 2 h at room temperature with 150 μl/well 1% BSA in PBS before loading samples (50 μl/well). Plates were then incubated overnight at 4°C and washed before adding 50 μl of 0.25 μg/ml biotinylated anti-mouse IgG2a Ab (Southern Biotechnology Associates, Birmingham, AL) to each well. Following a 2-h incubation at room temperature, plates were washed, incubated with alkaline phosphatase/streptavidin for 1 h at room temperature (50 μl/well at a concentration of 1:1000), and developed with p-nitrophenyl phosphate substrate dissolved in diethanolamine buffer. This ELISA was standardized with serum obtained from mice sensitized to OVA according to our Th1-polarized mucosal sensitization model and bled at day 11 of the protocol. Ig levels are expressed in units per milliliter relative to standard sera.

Summary:
1. IP-10 intervention resulted in a 4- to 10-fold increase in the level of IFN-γ in the BAL, while IL-4 was virtually undetectable and IL-13 was significantly attenuated; IL-5 content in the BAL was also reduced by about 40%, although this change did not reach statistical significance. Moreover, mice treated with IP-10 presented 2- to 3-fold higher levels of the proinflammatory chemokines MCP-1, MIP-1α, and RANTES in the BAL and produced three to four times more OVA-specific IgG2a, a Th1-affiliated Ig, than mice treated with GM-CSF alone or concurrently infected with GM-CSF and RDA control. 2. It is noteworthy that IP-10 treatment did not result in a statistically significant reduction in levels of OVA-specific IgE, nor did it alter expression of eotaxin, hallmarks of the eosinophilic response, an intriguing observation given the dramatic reduction in BAL, tissue, and peripheral blood eosinophilia in IP-10-treated animals. 3. The change was the consequence of a ∼50% increase in the number of activated CD8+ T cells and a parallel ∼50% reduction in activated CD4+ T cells in the airways of IP-10-treated mice compared with mice initially exposed to OVA in the context of GM-CSF and RDA; RDA intervention had no distinguishable effect on the lymphocyte profile following OVA rechallenge compared with mice receiving GM-CSF alone (data not shown).

References:
1: Wiley R, Palmer K, Gajewska B, Stämpfli M, Alvarez D, Coyle A, Gutierrez-Ramos J, Jordana M. Expression of the Th1 chemokine IFN-gamma-inducible protein 10 in the airway alters mucosal allergic sensitization in mice. J Immunol. 2001 Feb 15;166(4):2750-9. PubMed PMID: 11160341.