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USP42 (or Ubiquitin Specific Peptidase 42) is a ubiquitase protein that regulates transcriptional activity via its ability to deubiquitylate the histones that bind DNA within a cell’s nucleus. This prevents the cell’s transcriptional machinery from accessing gene promoter regions, leading to a reduction in the expression of specific genes.
USP42 is also known to target p53, stabilising p53 in response to cellular stress.
1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: USP42 Polyclonal Antibody (STJ96204)
2. What was used? Primary antibody: USP42 Polyclonal antibody (STJ96204) Provider: St John’s Laboratory Dilution ratio: 1:1000 Application: Western Blot Materials for validation: HeLa cells, 293T cells, bone marrow derived macrophages (BMDM) from mouse Click for product data sheet PDF
3. What was the protocol? Treatment of Materials Cells were cultured to 70-80% plating density. Cells were lysed in lysis buffer, supplemented with protease inhibitors (Roche). Proteins were reduced and denatured by heating at 100¡æ for 5 min with subsequent cooling before use. Gel Electrophoresis 30ug was loaded per lane Transfer Nitrocellulose membrane Blocking 1X TBST with 5% BSA for 1 hour Primary Antibody Application Overnight at 4¡ãC in 5% BSA in TBST Secondary Antibody Application After washing with TBST x3, membrane incubated with secondary antibody at room temperature for 1 hour Visualisation ECL kit
4. What was the protocol? Gel electrophoresis information£º7.5% polyacrylamide gel, constant amperes 20mA, 90min Transfer information£º0.2um NC membrane, 1 piece of gel (wet transfer), constant amperes 350mA 90min No. Antigen Loading amount Primary antibody Primary antibody dilution ratio Secondary antibody dilution ratio Target band KD Visualization time 1 293T cell lysate 30 ug STJ96204. USP42 antibody 1:1000 1:10000 ~150KD 20sec 2 HeLa cell lysate 30 ug 1:10000 ~150KD 20sec 3 BMDM cell lysate 20ug 1:10000 ~150KD 20sec
5. What were the results? Mock HA-USP42 200 116 97 66 kDa 293T Cell line 200 116 97 66 66 alpha-USP42 alpha-tubulin siUSP42sicontrol HeLa Cell line 200 116 97 66 alpha-USP42 alpha-tubulin Mouse BMDM
6. ¡°STJ96204 USP42 can detect the exogenous as well as endogenous USP42 protein in human and mouse cells.¡± What did the customer think? Antibody Specificity: Antibody Rating: Testing results were provided independently by Dr Nguyen Thi Hong Trang at Sungkyunkwan University. They stated that:
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