Anti-GPVI 抗体 [EPR25690-71] (ab302950)

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Negative control: human pancreas (human protein atlas database).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The identity of the lower MW band at approximately 45 kDa is unknown.

Exposure time:

Lane 1-3: 10 seconds

Lane 4-5: 48 seconds

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GPVI with ab302950 at 1/100 dilution (4.53 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Positive staining on megakaryocytes of mouse spleen. The section was incubated with ab302950 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Primary diluent was used instead of primary antibody, followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling GPVI with ab302950 at 1/100 dilution (4.53 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection). Negative control: no staining on mouse pancreas. The section was incubated with ab302950 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen (fresh) tissue labeling GPVI with ab302950 at 1/500 (9.06 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing positive staining on megakaryocytes of mouse spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab302950 for 60 mins at room temperature. The section was then mounted using Fluoromount^®. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 (2 µg/ml) dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse pancreas (fresh) tissue labeling GPVI with ab302950 at 1/500 (9.06 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). Negative control: confocal image showing positive staining on mouse pancreas (PMID:10961879). The nuclear counterstain was DAPI (Blue). The section was incubated with ab302950 for 60 mins at room temperature. The section was then mounted using Fluoromount^®.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbedat 1/1000 (2 µg/ml) dilution.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEL (human erythroleukemia erythroblast) cells labeling GPVI with ab302950 at 1/100 (4.53 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green). Confocal image showing increased membranal and cytoplasmic staining in HEL cells treated with Phorbol-12-myristate-13-acetate (10 nM) for 48 hours. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.