CCL25 Antibody (OASA07519)

Catalog#: OASA07519

• Size: 0.1mg

• Description of Target: RABBIT ANTI HUMAN CCL25
• 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
• Immunogen: The immunogen for anti-CCL25 antibody: recombinant human CCL25
• Product Format: Purified IgG - lyophilised
• Isotype: Polyclonal IgG
• Predicted Homology Based on Immunogen Sequence: Human
• Applications Info: ELISA : This antibody may be used in an indirect ELISA or as a capture reagent in a sandwich ELISA with This product B as the detection 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 1.0mg/ml after reconstitution
  
  Buffer Solutions: Phosphate buffered saline pH7.2
• Reconstitution and Storage: Reconstitution: Reconstitute with 0.1ml distilled water.
  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 mixed after reconstitution. For l

Product Protocol: CCL25 antibody used to evaluate hepatic endothelial ccl25 mediates the recruitment of ccr9+ gut-homing lymphocytes to the liver in primary sclerosing cholangitis (OASA07519)

Specificity: CCL25
Product Page: CCL25 antibody (OASA07519)
Antibody Pair Available: OASA07519 Capture OASA07520 Detection
Experiment Type: Immunohistochemistry and Dual-Color Coimmunofluorescence

Protocol:
6-μm cryostat sections were fixed in acetone for 10 min. 15 cases of PSC and 5 cases each of normal liver, primary biliary cirrhosis (PBC), and alcoholic liver disease were studied. Sections were incubated with 20% goat serum before rabbit anti–human CCL25 polyclonal antibody (P134; 1 μg/ml; PeproTech) or mouse anti–human CXCL12 mAb (MAB350; 8 μg/ml; R&D Systems) overnight at 4°C. Control sections were incubated without primary antibody or rabbit immunoglobulin. Sections were incubated for 20 min with biotinylated secondary antibodies followed by streptavidin–horseradish peroxidase complex developed with diaminobenzidine and counterstained with hematoxylin.

Sections for dual immunofluorescence were incubated with 20% goat/rabbit serum for 30 min before primary antibodies raised against CD68, CD31, Cytokeratin 19 (DakoCytomation), LYVE-1 (Upstate Biotechnology), or CCL25. Control sections were incubated without primary antibody. Sections were stained with goat anti–rabbit FITC (Southern Biotechnology Associates, Inc.) and goat anti–mouse IgG1 or IgG2a Texas red (Southern Biotechnology Associates, Inc.) secondary antibodies and nuclei counterstained with DAPI. Immunofluorescence was assessed using AxioVision software (Carl Zeiss MicroImaging, Inc.).

Summary:
1. A functional role for CCR9 in PSC was supported by the strong CCL25 expression detected in the liver in PSC, but not in other chronic inflammatory diseases. CCL25 was detected in portal dendritic cells/macrophages and hepatic sinusoidal endothelium at areas of interface hepatitis in PSC, but was absent from other chronic inflammatory diseases. 2. The results demonstrate for the first time aberrant expression of CCL25 outside the gut or thymus associated with infiltration of CCR9+ T cells into the liver. The ability of CCL25 to activate migration and α4β7-mediated adhesion of LILs to MAdCAM-1 suggests that CCL25 and MAdCAM-1 cooperate in the recruitment of mucosal lymphocytes to the liver in PSC.

References:
1: Eksteen B, Grant AJ, Miles A, Curbishley SM, Lalor PF, Hübscher SG, Briskin M, Salmon M, Adams DH. Hepatic endothelial CCL25 mediates the recruitment of CCR9+ gut-homing lymphocytes to the liver in primary sclerosing cholangitis. J Exp Med. 2004 Dec 6;200(11):1511-7. Epub 2004 Nov 22. PubMed PMID: 15557349; PubMed Central PMCID: PMC2211943.