6X His tag [3H2201]

3H2201

PAX6 transcription factor linked to 6-histidine tag

6X His tag

From hybridoma culture supernatant Storage

For ELISA: Use at an assay dependent dilution. For IHC-Fr: Use at an assay dependent dilution. For IP: Use at an assay dependent dilution. For WB: Use at a dilution of 1:250-1:500. Not tested in other applications. Optimal dilutions/concentrations should be determined by the end userMyeloma: NS1Protocols from PublicationsCitation: 1: Kolisek M Launay P Beck A Sponder G Serafini N Brenkus M Froschauer EM Martens H Fleig A Schweigel M. SLC41A1 is a novel mammalian Mg2+ carrier. J Biol Chem. 2008 Jun 6; 283(23):16235-47. Epub 2008 Mar 25. PubMed PMID: 18367447; PubMed Central PMCID: PMC2414286. Experiment Name: Use of His6 tag antibody in immunoprecipitation and western Blot AnalysisExperiment Background: 1. SLC41A1 is a bona fide magnesium transporter. Most importantly overexpressed SLC41A1 provide HEK293 cells with an increased magnesium efflux capacity. 2. hSLC41A1 expression was induced by addition of 0. 05 mmol liter-1 isopropyl b-D-thiogalactopyranoside (IPTG) to the growth media. Experiment Steps: 1. Extract total proteins from 250 ml of the bacterial culture (-IPTG or +IPTG as indicated) using trichloroacetic acid/acetone. 2. Isolate proteins of the membrane fraction using the ProteoExtractTM Partial Bacterial Proteome Extraction Kit (Calbiochem La Jolla CA). 3. Immunoprecipitate His-tagged hSLC41A1 from the membrane protein fraction with a His6 tag antibody ( Biotech San Diego CA). 4. Separate protein samples by SDS-PAGE utilizing 12. 5% polyacrylamide gels blot and label with His6 tag antibody and goat anti-mouse (GAM)-HRP (Molecular Probes Eugene OR) or GAM-k-HRP (SBA Birmingham AL) antibodies. 5. Visualize antibody binding using the ChemilmagerTM 5500 (Alpha Innotech) or AGFA Cronex 5 medical x-ray films developed with the Curix 60 (AGFA) This product recognises proteins containing the motif HHHHHH and is therefore of value in detecting proteins containing histidine tags. The antibody may be used to track the purification of proteins expressed in bacteria yeast or insect cells.