Anti-GRP94 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody (AB3674)

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.

• ICC/IF

AbReview5086****

Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody (AB3674)

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).

This image is courtesy of an anonymous Abreview

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody (AB3674)

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

• WB

Unknown

Western blot - Anti-GRP94 antibody (AB3674)

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

All lanes:

Western blot - Anti-GRP94 antibody (ab3674) at 1/2000 dilution

All lanes:

Cell lysates prepared from mouse fibroblasts

Predicted band size: 92 kDa

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• WB

AbReview25266****

Western blot - Anti-GRP94 antibody (AB3674)

All lanes:

Western blot - Anti-GRP94 antibody (ab3674) at 1/2000 dilution

Lane 1:

Whole cell lysate prepared from murine 3T3 cells at 50 µg

Lane 2:

Whole cell lysate prepared from murine 3T3 cells, treated with 2.5 ug/ml Tunicamycin 10 hours at 50 µg

Secondary

All lanes:

HRP conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 92 kDa

Observed band size: 94 kDa

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Exposure time: 30s

Image courtesy of an anonymous Abreview.