Human USP5 siRNA (L-006095-00) were obtained from Dharmacon (Lafayette, CO, USA), USP5 shRNA in pLKO.1 vector targeting mouse USP5 were purchased from Sigma (USP5 shRNA: TRCN0000030737, USP5 shRNA-B: TRCN0000030738) (St. Louis, MO, USA). Recombinant PD-L1 protein (NBP1-98984) was purchased from Novus Biologicals (Centennial, CO, USA). Flag-PD-L1 and myc-USP5 plasmids were obtained from GenScript (Piscataway, NJ). Anti-USP5 (sc-390943), anti-HA (sc-57592), anti-β-Actin (sc-47778) and anti-Flag (sc-807) antibodies were purchased from Santa Cruz (Santa Cruz, CA). Anti-PD-L1 antibody was purchased from Cell Signaling Technology (Danvers, MA). Mouse PD-L1 cDNA was purchased from Sino Biological Inc. (Beijing, China).
Anti flag sc 807
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-Flag (sc-807) is a mouse monoclonal antibody that binds to the FLAG epitope tag. FLAG is a commonly used short peptide tag that can be attached to proteins to facilitate detection and purification. The Anti-Flag (sc-807) antibody can be used to detect and immunoprecipitate FLAG-tagged proteins.
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Lab products found in correlation
3 protocols using anti flag sc 807
1
Regulation of PD-L1 by USP5
2
Zfp127 ChIP-qPCR on Oct4 promoter
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A chromatin immunoprecipitation (ChIP) assay was performed as described previously (19 (link), 28 (link)). In brief, Zfp127 overexpression was performed in NIH-3T3 and MEFs as mentioned above. 1 × 106 cells of Zfp127-overexpressing NIH-3T3 and MEFs were rinsed and treated with 1% formaldehyde for 20 min at room temperature. Samples were then sonicated on ice and incubated with the antibodies; anti-FLAG (sc-807)(Santa Cruz, USA) and anti-IgG antibody at 4°C overnight. Immunoprecipitated DNA was analyzed using real-time PCR. The following primers were used to amplify the primer of the Oct4 promoter; forward, 5′-GGAGGTGCAATGGCTGTCTTGTCC-3′, and reverse, 5′-CTGCCTTGGGTCACCTTACACCTCAC-3′.
3
Yeast Two-Hybrid System for Protein-Protein Interactions
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Yeast vectors, Y2H gold strain, Aureobasidin A (630466) and X-α gal (630463) were from Clontech (Mountain View, California, USA). Mav203 strain was from Life Technologies (Carlsbad, California, USA). Amino acid supplements, 3-Amino-1, 2, 4-triazole (3-AT, A8056), IPTG (I6758) were from Sigma-Aldrich (Saint Louis, Missouri, USA). Anti-His (SC-57598), anti-myc (SC-789) and anti-Flag (SC-807) antibodies were from Santa Cruz Biotechnology (Dallas, Texas, USA). Lysozyme (10153516103) was from Roche (Indianapolis, Indiana, USA). Ni NTA agarose (25215) and Glutathione agarose (16100) were from Thermo Scientific (Waltham, Massachusetts, USA). Peptide affinity purified rabbit polyclonal antibody against the macro domain (X) of HEV was generated at Genscript (Piscataway Township, New Jersey, USA) using KLH (Keyhole Limpet hemocyanin) conjugated peptide antigens. Peptide sequence is: CPDYRLEHNPKRLEA. Pre immune serum was collected from the animals prior to immunization. ELISA using peptides as antigen and pre immune serum as negative controls confirmed their titres to be >1:512,000. In competition assays, corresponding peptides could block the binding of cognate antibodies. Functionality of the antibody in western was validated by testing its ability to detect the purified X-domain (from bacteria) and the X-domain expressed in Huh 7 cells.
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