||Recombinant Human CCR5, fused to Flag-tag, was expressed in HEK293T cell.
||This gene encodes a member of the beta chemokine receptor family, which is predicted to be a seven transmembrane protein similar to G protein-coupled receptors. This protein is expressed by T cells and macrophages, and is known to be an important co-receptor for macrophage-tropic virus, including HIV, to enter host cells. Defective alleles of this gene have been associated with the HIV infection resistance. The ligands of this receptor include monocyte chemoattractant protein 2 (MCP-2), macrophage inflammatory protein 1 alpha (MIP-1 alpha), macrophage inflammatory protein 1 beta (MIP-1 beta) and regulated on activation normal T expressed and secreted protein (RANTES). Expression of this gene was also detected in a promyeloblastic cell line, suggesting that this protein may play a role in granulocyte lineage proliferation and differentiation. This gene is located at the chemokine receptor gene cluster region. Two transcript variants encoding the same protein have been found for this gene.
||1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer.1 vial of 100 µg empty vector transfected control cell lysate in RIPA buffer.1 vial of 250ul 2xSDS Sample Buffer (4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromphenol blue, 100mM DTT).
||Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4), and then centrifuged to clarify the lysate. Protein concentration was measured by BCA kit. Cell lysates were aliquoted and stored at -20°C before shipping.
||The lysate is shipped with dry ice. Upon receiving, store the sample at -20°C. Lysate samples are stable for 12 months from date of receipt when stored at -20°C. Avoid repeated freeze-thaw cycles.Lysate samples can be diluted with 2xSDS Sample Buffer provided. After dilution, the protein sample should be aliquoted and stored at -20°C for long term storage. Prior to SDS-PAGE fractionation, boil the lysate for 5 minutes.