CCL25 Antibody (OASA07520)

Catalog#: OASA07520

• Size: 50ug

• Description of Target: RABBIT ANTI HUMAN CCL25:Biotin
• Gene Symbol: CCL25
• Alias Symbols: TECK; Ckb15; SCYA25; MGC150327; CCL25
• Protein Accession#: NP_005615.2
• Swissprot Id: O15444
• Host: Rabbit
• Protein Size (# AA): 150
• Specificity: CCL25
• Application: ELISA, WB
• Conjugation: Biotin
• Immunogen: The immunogen for anti-CCL25 antibody: recombinant human CCL25
• Product Format: Purified IgG conjugated to Biotin - lyophilised
• Isotype: Polyclonal IgG
• Applications Info: ELISA : This antibody may be used in an indirect ELISA or as a detection reagent in a sandwich ELISA with OASA07519 as the capture antibody.
  Preservative Stabilisers: None present
  
  Antiserum Preparation: Antiserum to human CCL25 were raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
  Approx Protein Conc: IgG concentration 0.1mg/ml after reconstitution
  
  Buffer Solutions: Phosphate buffered saline pH7.2
• Reconstitution and Storage: Reconstitution: Reconstitute with 0.5ml 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: CCL25 antibody used to evaluate human g protein-coupled receptor gpr-9-6/cc chemokine receptor 9 is selectively expressed on intestinal homing t lymphocytes, mucosal lymphocytes, and thymocytes and is required for thymus-expressed chemokine-mediated chemotaxis (OASA07520)

Specificity: CCL25
Product Page: CCL25 antibody (OASA07520)
Antibody Pair Available: OASA07519 Capture OASA07520 Detection
Experiment Type: Binding Assay with Biotinylated TECK

Protocol:
4 × 105 cells in 25 μl of PBS with 10% rabbit serum, 1% FCS, and 0.1% sodium azide (assay buffer) were preincubated with either assay buffer (no treatment), 1,000 nM TECK/MCP-1, or 50 μg/ml 3C3/IgG2b for 30 min. 25 μl of biotinylated TECK at a final concentration ranging up to 100 nM was then added, and after 1 h, the cells were washed twice with 200 μl of wash buffer (PBS with 1% FCS and 0.1% azide) and resuspended in 25 μl of Wallac Eμ-labeled streptavidin diluted 1:400 in assay buffer. After 30 min at room temperature, the cells were washed in wash buffer and transferred to a 96-well solid white nonspecific binding plate (3600; Corning) and, after spinning to remove supernatant, 200 μl of Wallac Enhancement Solution was added. The fluorescence was counted on a multilabel counter (model 1420; Wallac) at 340/613 nm and reported in relative fluorescence units (RFUs).

Summary:
1. Although tyrosine-iodinated TECK did not bind to GPR-9-6+ Molt-4 cells, biotinylated TECK bound in a dose-dependent and saturable manner. In contrast, biotinylated TECK did not bind to GPR-9-6− CEM cells. 2. Binding of biotinylated TECK to Molt-4 cells was inhibited by either 500 nM TECK or by the anti–GPR-9-6 mAb 3C3, but not by either 500 nM MCP-1 or an IgG2b control mAb. 3. Transcripts of GPR-9-6 and TECK are present in both small intestine and thymus. Importantly, the expression profile of GPR-9-6 correlates with migration to TECK of blood T lymphocytes and thymocytes.

References:
1: Zabel BA, Agace WW, Campbell JJ, Heath HM, Parent D, Roberts AI, Ebert EC, Kassam N, Qin S, Zovko M, LaRosa GJ, Yang LL, Soler D, Butcher EC, Ponath PD, Parker CM, Andrew DP. Human G protein-coupled receptor GPR-9-6/CC chemokine receptor 9 is selectively expressed on intestinal homing T lymphocytes, mucosal lymphocytes, and thymocytes and is required for thymus-expressed chemokine-mediated chemotaxis. J Exp Med. 1999 Nov 1;190(9):1241-56. PubMed PMID: 10544196; PubMed Central PMCID: PMC2195678.