TBC1D4 [p Thr642] Antibody (JE60-97)

Western Blot: TBC1D4 [p Thr642] Antibody (JE60-97) [NBP3-32749]

Western Blot: TBC1D4 [p Thr642] Antibody (JE60-97) [NBP3-32749] - Western blot analysis of TBC1D4 on different lysates with Rabbit anti-TBC1D4 antibody (NBP3-32749) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate
Lane 2: NIH/3T3 starve overnight then treated with 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes cell lysate

Lysates/proteins at 20 ug/Lane.

Predicted band size: 147 kDa
Observed band size: 160 kDa

Exposure time: 25 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32749) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/ Immunofluorescence: TBC1D4 [p Thr642] Antibody (JE60-97) [NBP3-32749]

Immunocytochemistry/ Immunofluorescence: TBC1D4 [p Thr642] Antibody (JE60-97) [NBP3-32749] - Immunocytochemistry analysis of NIH/3T3 cells starved overnight then treated with or without 100nM Calyculin A for 50 minutes add 100ng/mL insulin for 20 minutes labeling TBC1D4 with Rabbit anti-TBC1D4 antibody (NBP3-32749) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TBC1D4 antibody (NBP3-32749) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.