Immunostaining of mouse embryonic fibroblasts with ab3674. (See Nakamura et al. 2001).

Anti-GRP94 antibody (ab3674) at 1/2000 dilution + Cell lysates prepared from mouse fibroblasts


Western blot of mouse fibroblasts using ab3764 at 1/2000.

ab3674 at a 1/100 dilution staining human renal cell carcinoma cells by ICC/IF. The cells were permeabilized with 0.2% triton X100 and then blocked with 1%BSA, 4% goat serum prior to incubation with the primary 1antibody for 30 min at 37 °C. Bound antibody was detected using an Alexa fluor ® 488 Goat anti-Rabbit IgG (H+L).

See Abreview

Ab3674 (1/1000) staining GPR94 in human stomach using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of peptic and parietal cells of the gastric gland.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

All lanes : Anti-GRP94 antibody (ab3674) at 1/2000 dilution

Lane 1 : Whole cell lysate prepared from murine 3T3 cells
Lane 2 : Whole cell lysate prepared from murine 3T3 cells, treated with 2.5 ug/ml Tunicamycin 10 hours

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : HRP conjugated goat anti-rabbit IgG at 1/2000 dilution

Developed using the ECL technique.

Observed band size: 94 kDa why is the actual band size different from the predicted?


Exposure time: 30 seconds

See Abreview