Chemokine (C-C Motif) Receptor 9 (CCR9) Antikörper

Antigen: Chemokine (C-C Motif) Receptor 9 (CCR9)

Synonyme: GPR28, CDw199, GPR-9-6, D6, hD6, CCR9, CCR10, CMKBR9, MGC126678, MGC138250

Klonalität: Monoklonal (7E07)

Wirt: Ratte

Reaktivität: Maus

Applikation: Immunhistochemie (Gefrierschnitte) (IHC (fro)), Durchflusszytometrie (FACS), Immunfluoreszenz (IF)

Produktnummer: ABIN316578

Produktbeschreibung

Produktmerkmale: Synonyms: C-C chemokine receptor type 9, C-C CKR-9, CC-CKR-9, CCR-9, GPR-9-6, CMKBR9, GPR28,G-protein coupled receptor 28

Weitere Bezeichnung: CDw199 / CCR9

Swiss-Prot: Q9WUT7

Immunogen: Peptide comprising amino acids 3-22 of mouse CCR9

Kreuzreaktivität: Maus

Isotyp: IgG2b  (Passende Sekundärantikörper)

Klon: 7E07

Beschreibung: The protein encoded by this gene is a member of the beta chemokine receptor family. It ispredicted to be a seven transmembrane protein similar to G protein coupled receptors. Chemokines and their receptors are key regulators of the thymocytes migration andmaturation in normal and inflammation conditions. This gene is expressed in a range oftissues and hemopoietic cells. The expression of this receptor in lymphatic endothelialcells and overexpression in vascular tumors suggested its function in chemokine-drivenrecirculation of leukocytes and possible chemokine effects on the development andgrowth of vascular tumors. This receptor appears to bind the majority of beta-chemokinefamily members, however, its specific function remains unknown. The specific ligand ofthis receptor is CCL25. It has been found that this gene is differentially expressed by Tlymphocytes of small intestine and colon, suggested a role in the thymocytes recruitmentand development that may permit functional specialization of immune responses indifferent segment of the gastrointestinal tract. This gene is mapped to chromosome3p21. 3, a region that includes a cluster of chemokine receptor genes. Two alternativelyspliced transcript variants have been described.

Spezifität: The antibody recognizes murine CCR-9. Other species not tested.

Anwendungen

Protokoll: Immunofluorescence protocol - Formaldehyde fixation1. Collect cells from T. c. unit and remove media from petri dish using suction. 2. Wash with 1x PBS and remove. 3. Incubate cells in pre-warm (37°C) Paraformaldehyde for 12 minutes at room temperatureon an orbital shaker. 4. Remove PFA and incubate in 0. 5% Triton X-IOO in 1x PBS for 5 minutes at roomtemperature. 5. Prepare blocking reagent, this is also the antibody diluent. 6. Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker. 7. Block with 1 % NCS and 1x PBS for 30 minutes at room temperature. 8. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. 9. Wash cells 2x with lx PBS at room temperature and air dry briefly. 10. Incubate with primary antibody for 1 hr at room temperature in the dark in stainingchambers. During this time prepare the secondary antibody. 11. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)12. Incubate with secondary antibody for 1 hour at room temperature in the dark in stainingchambers. 13. Wash cells 5x with 1x PBS. 14. Mount in Dapi. Solutions (prepare fresh the same day of staining). 1x Phosphate buffered saline. Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS). Fixation solution: 3. 5% Paraformaldehyde. 1. 75g PFA in 20 ml d. H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C untildissolved. Add 4 drops I N HCI and check pH indicator strip. pH should be 7. 4. Completevolume with d. H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips. Immunofluorescence protocol - Methanol/acetone fixation1. Collect cells from T. C. unit and remove media from petri dish using suction. 2. Wash with 1x PBS and remove. 3. Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice. 4. Prepare blocking reagent, this is also the diluent for the antibodies. 5. Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbitalshaker. 6. Block with 1% NCS and Ix PBS for 30 minutes at RT. 7. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. 8. Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes. 9. Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. Duringthis time prepare secondary antibody. 10. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)11. Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers. 12. Wash cells 5x with 1x PBS. 13. Mount in Dapi.

Applikationshinweise: Immunohistochemistry on frozen sections. Flow cytometry. Immunofluorescence. Recommended dilution: 1: 20. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Reinigung: Supernatant

Lagerung: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Shelf life: one year from despatch.

Forschungsgebiet: Chemokine, Rezeptoren

Beschränkungen: Nur für Forschungszwecke einsetzbar