Anti-ASIC3 抗体 [EPR26557-80] - BSA and Azide free (ab302777)

This data was developed using ab302776, the same antibody clone in a different buffer formulation. 

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

The MW is consistent with what has been described in the literature (PMID: 17012229)

Negative control: rat: hippocampus, spinal cord, kidney, testis.(PMID:11872753)

Samples are non-boiled as boiling may cause protein aggregates.

This blot was developed using a high sensitivity ECL substrate in lane 6 and 7. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

Exposure time: 

Lane 1-5: 180 seconds

Lane 6 and 7: 37 seconds. 

This data was developed using ab302776, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse dorsal root ganglion tissue labeling ASIC3 with ab302776 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on mouse dorsal root ganglion. The section was incubated with ab302776 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 LeicaDS9800 (BOND™ Polymer Refine Detection).

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

This data was developed using ab302776, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded rat dorsal root ganglion tissue labeling ASIC3 with ab302776 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on rat dorsal root ganglion (PMID: 9707631). The section was incubated with ab302776 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 LeicaDS9800 (BOND™ Polymer Refine Detection).

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

This data was developed using ab302776, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling ASIC3 with ab302776 at 1/5000 dilution (0.1 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Negative control: no staining on mouse testis was observed (PMID:11872753). The section was incubated with ab302776 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 LeicaDS9800 (BOND™ Polymer Refine Detection).

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

This data was developed using ab302776, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK 293T (human embryonic kidney epithelial cell) cells labeling ASIC3 with ab302776 at 1/4000 dilution (0.124 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing positive staining in HEK-293T cells transfected with a mouse ASIC3 expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ICC/IF has not been tested with endogenous material. Myc-Tag Mouse mAb (Alexa Fluor^® 647) was used to counterstain tubulin at 1/100 dilution (0.38 µg/ml) (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 dilution (2 µg/ml).

This data was developed using ab302776, the same antibody clone in a different buffer formulation.

ASIC3 was immunoprecipitated from 0.35 mg rat dorsal ganglion tissue lysate with ab302776 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302776 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: rat dorsal ganglion tissue lysate 10 μg (Inset)

Lane 2: ab302776 IP in rat dorsal ganglion tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302776 in rat dorsal ganglion tissue lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Samples are non-boiled as boiling may cause protein aggregates in lane 1.

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.