Anti-TRPC3 antibody [EPR25056-56]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPC3 antibody [EPR25056-56] (AB300142)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T cells transfected with a TRPC3 expression vector containing a his tag and (B) HEK-293T cells transfected with empty vector containing a his tag. tissue labeling TRPC3 with ab300142 at 1/500 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Cytoplasmic staining on (A) HEK-293T transfected with a TRPC3 expression vector; nearly no staining on (B) HEK-293T transfected with empty vector.

The section was incubated with ab300142 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPC3 antibody [EPR25056-56] (AB300142)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling TRPC3 with ab300142 at 1/100 dilution.

Negative control : no staining on mouse liver.

The section was incubated with ab300142 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 solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPC3 antibody [EPR25056-56] (AB300142)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling TRPC3 with ab300142 at 1/100 dilution.

Positive staining on mouse cerebellum.

The section was incubated with ab300142 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 solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPC3 antibody [EPR25056-56] (AB300142)

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling TRPC3 with ab300142 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining on rat cerebellum.

The section was incubated with ab300142 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Lab

Western blot - Anti-TRPC3 antibody [EPR25056-56] (AB300142)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

The molecular weight is consistent to literature (PMID : 15834157).

Low express tissue : lung (PMID : 21931279).

All lanes:

Western blot - Anti-TRPC3 antibody [EPR25056-56] (ab300142) at 1/1000 dilution

Lane 1:

Mouse cerebellum tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse lung tissue lysate at 20 µg

Lane 4:

Rat cerebellum tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat lung tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 105 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-TRPC3 antibody [EPR25056-56] (AB300142)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining.

All lanes:

Western blot - Anti-TRPC3 antibody [EPR25056-56] (ab300142) at 1/1000 dilution

Lane 1:

HEK-293 transfected with mouse TRPC3 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

HEK-293 transfected with mouse TRPC7 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

HEK-293 transfected with mouse TRPC6 (WT) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 105 kDa

false

Exposure time: 6s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPC3 antibody [EPR25056-56] (AB300142)

Immunohistochemical analysis of paraffin-embedded Panel A HEK-293T cells labeling TRPC3 with ab300142 at 1/500 dilution followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Cytoplasmic staining on (A) HEK-293T transfected with a TRPC3 expression vector; nearly no staining on (B) HEK-293T transfected with empty vector. The section was incubated with ab300142 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.