Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labelling GABA A Receptor alpha 6 with ab300069 at 1/500 dilution (1.244 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on human cerebellum. The section was incubated with ab300069 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human cerebellum staining of Calbindin with ab108404 at a 1/1000 dilution, DGKZ/DGK-zeta with ab239081 at 1/5000 dilution, and GABA A Receptor alpha 6 with ab300069 at a 1/500 dilution, followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-GABA A Receptor alpha 6 (gray; Opal™690), anti-DGKZ/DGK-zeta (green; Opal™520) and anti-Calbindin (magenta; Opal™570) on human cerebellum.
Panel B : anti-Calbindin stained on all layers of cerebellum.
Panel C : anti-DGKZ/DGK-zeta stained on Purkinje cells.
Panel D : anti-GABA A Receptor alpha 6 stained on the granular layer.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300069, ab239081, and ab108404 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

Fluorescence multiplex immunohistochemical analysis of mouse cerebellum (formalin-fixed paraffin-embedded section).

Panel A : merged staining of anti-GABRA6 (magenta; Opal™690), anti-KCNA (green; Opal™520) and anti-DGKZ (gray; Opal™570) on mouse cerebellum.
Panel B : anti-GABRA6 staining granule cells in mouse cerebellum.
Panel C : anti-KCNA staining basket cells in mouse cerebellum.
Panel D : anti-DGKZ staining Purkinje cells in mouse cerebellum.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300069 at a 1/500 dilution, ab313624 at a 1/5000 and ab239080 at a 1/5000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labelling GABA A Receptor alpha 6 with ab300069 at 1/500 dilution (1.244 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on mouse cerebellum. The section was incubated with ab300069 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.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labelling GABA A Receptor alpha 6 with ab300069 at 1/500 dilution (1.244 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on rat cerebellum. The section was incubated with ab300069 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-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum (fresh) tissue labeling GABA A Receptor alpha 6 with ab300069 at 1/500 (1.244 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 μg/mL) dilution (Green). Positive staining on rat cerebellum is observed. 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 μ/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh) tissue labeling GABA A Receptor alpha 6 with ab300069 at 1/500 (1.244 μg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 2 μg/mL dilution (Green). Positive staining on mouse cerebellum is observed. 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 μ/mL dilution.

• WB

Supplier Data

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

5% NFDM/TBST was used as blocking and diluting buffer.

Low expression：Heart (PMID : 29467616).

All lanes:

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] (<a href='/en-us/products/primary-antibodies/gaba-a-receptor-alpha-6-antibody-epr25322-129-ab300069'>ab300069</a>) at 1/1000 dilution

Lane 1:

Human hypothalamus tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 51 kDa

Observed band size: 57 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

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

Low expression tissue : heart (PMID : 29467616).

This blot was developed using a high ECL substrate.

Sometimes a non-specific band near the target signal can be observed.

All lanes:

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] (<a href='/en-us/products/primary-antibodies/gaba-a-receptor-alpha-6-antibody-epr25322-129-ab300069'>ab300069</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human hypothalamus tissue lysate at 20 µg

Lane 3:

Human heart tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Mouse cerebellum tissue lysate at 20 µg

Lane 6:

Mouse heart tissue lysate at 20 µg

Lane 7:

Rat brain tissue lysate at 20 µg

Lane 8:

Rat cerebellum tissue lysate at 20 µg

Lane 9:

Rat heart tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP), minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 51 kDa

Observed band size: 57 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

5% NFDM/TBST was used as blocking and diluting buffer.

All lanes:

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] (<a href='/en-us/products/primary-antibodies/gaba-a-receptor-alpha-6-antibody-epr25322-129-ab300069'>ab300069</a>) at 1/1000 dilution

All lanes:

Mouse brain 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

Predicted band size: 51 kDa

Observed band size: 57 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] - BSA and Azide free (AB300070)

This data was developed using ab300069, the same antibody clone in a different buffer formulation.

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

Low expression : Heart (PMID : 29467616)

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.

All lanes:

Western blot - Anti-GABA A Receptor alpha 6 antibody [EPR25322-129] (<a href='/en-us/products/primary-antibodies/gaba-a-receptor-alpha-6-antibody-epr25322-129-ab300069'>ab300069</a>) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat cerebellum tissue lysate at 20 µg

Lane 3:

Rat heart tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L) at 1/2000 dilution

Predicted band size: 51 kDa

Observed band size: 57 kDa

true

Exposure time: 180s