Anti-IDH3A antibody (ab58641)

Western blot - IDH3A antibody (ab58641)

Anti-IDH3A antibody (ab58641) at 1.25 µg/ml + Jurkat cell lysate at 10 µg

Secondary
HRP conjugated anti-Rabbit IgG at 1/50000 dilution

Predicted band size : 40 kDa
Observed band size : 40 kDa

Western blot - IDH3A antibody (ab58641)

All lanes : Anti-IDH3A antibody (ab58641) at 1/10000 dilution (For 16 hours at 4°C)

Lane 1 : Mouse adipocyte whole cell lysate
Lane 2 : Mouse adipocyte whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/10000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 40 kDa
Observed band size : 40 kDa


Exposure time : 1 second

This image is courtesy of an anonymous Abreview

Immunoprecipitation - Anti-IDH3A antibody (ab58641)

IDH3A was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to IDH3A and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab58641.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 38kDa; IDH3A.

Immunohistochemistry (Paraffin-embedded sections) - IDH3A antibody (ab58641)

ab58641 at a 4.0µg/ml dilution staining IDH3A in human skeletal muscle cells. Arrows indicate positively labelled skeletal muscle cells.

Immunocytochemistry/ Immunofluorescence-IDH3A antibody(ab58641)

ICC/IF image of ab58641 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab58641, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.