Heat shock 60KD protein(HSP60) is a member of the chaperonin class of protein factors and the nuclear encoded mitochondrial HSP60 is required for the assembly into oligomeric complexes of proteins imported into the mitochondrial matrix. HSP60 is linked head to head comprising approximately 17 kb and consist of 12 exons. HSP60 is a self-molecule, it can downregulate adaptive immune responses by upregulating Tregs innately through TLR2 signaling.
Western blot : 2-4µg/ml, Immunohistochemistry(Paraffin-embedded Section) : 4-8µg/ml
Figure 1. Western blot analysis of HSP60 using anti-HSP60 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human U87 whole cell lysates, Lane 3: rat brain tissue lysates, Lane 4: mouse Neuro-2a whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-HSP60 antigen affinity purified monoclonal antibody at 2 µg/mL overnight at 4C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for HSP60 at approximately 65 kDa. The expected band size for HSP60 is at 65 kDa.
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