HSPA4 Antibody / Heat shock protein family A member 4

Western Blot: 0.25-0.5 µg/mL, Immunohistochemistry: 2-5 µg/mL, Immunocytochemistry: 5 µg/mL, Immunofluorescence: 5 µg/mL, Flow cytometry: 1-3ug/million cells

After reconstitution, the HSPA4 antibody can be stored for up to one month at 4°C. For long-term, aliquot and store at -20°C. Avoid repeated freezing and thawing.

Western blot analysis of HSPA4 using anti-HSPA4 antibody. Lane 1: human 293T whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: rat brain tissue lysates, Lane 4: rat C6 whole cell lysates, Lane 5: mouse brain tissue lysates, Lane 6: mouse NIH/3T3 whole cell lysates, Lane 7: mouse Raw264.7 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 rabbit anti-HSPA4 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. A dominant band is observed at approximately 105-110 kDa, which is the expected migration pattern for HSPA4. Although the theoretical molecular weight of HSPA4 is ~94 kDa, members of the Hsp110 family, including HSPA4, are well known to migrate 10-20 kDa heavier due to acidic domains, extended disordered regions, and phosphorylation, which reduce SDS binding and cause anomalous gel mobility.