Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue labelling Eph receptor B2 with ab252935 at 1/500 dilution (B), GSDMC2 + GSDMC3 with ab229896 at 1/500 dilution (C) and Trefoil Factor 3 with ab300427 at 1/500 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Eph receptor B2 (green; Opal™520), anti-GSDMC2/3 (magenta; Opal™690) and anti-Trefoil Factor 3 (gray; Opal™570) on mouse colon.

Panel B : anti-Eph receptor B2 staining stem cells in mouse colon.

Panel C : anti-GSDMC2/3 staining epithelium in mouse colon.

Panel D : anti-Trefoil Factor 3 staining goblet cells in mouse colon.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab252935, ab229896 and ab300427 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue labelling Eph receptor B2 with ab252935 at 1/500 dilution (B), GSDMC2 + GSDMC3 with ab229896 at 1/500 dilution (C) and Serotonin with ab315150 at 1/500 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Eph receptor B2 (green; Opal™520), anti-GSDMC2/3 (magenta; Opal™690) and anti-Serotonin (gray; Opal™570) on mouse stomach.

Panel B : anti-Eph receptor B2 staining stem cells in mouse stomach.

Panel C : anti-GSDMC2/3 staining epithelium in mouse stomach.

Panel D : anti-Serotonin staining enterochromaffin cells in mouse stomach.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab252935, ab229896 and ab315150 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling GSDMC2+GSDMC3 with ab229896 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplamic staining on epithelial cells in mouse stomach (PMID : 17350798). The section was incubated with ab229896 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse colon tissue labeling GSDMC2+GSDMC3 with ab229896 at 1/200 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on epithelial cells in mouse colon is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse stomach tissue labeling GSDMC2+GSDMC3 with ab229896 at 1/200 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on epithelial cells in mouse stomach is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GSDMC2+GSDMC3 with ab229896 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : No staining on mouse spleen (PMID : 17350798). The section was incubated with ab229896 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse colon tissue staining GSDMC2 + GSDMC3 with ab229896 at a 1 : 2000 (0.25 ug/ml) dilution; SARM1 with ab309195 at 1 : 500 (0.964 ug/ml) dilution and MUC2 with ab272692 at 1 : 2000 (0.252 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-SARM1 (green; Opal^™520), anti-MUC2 (gray; Opal^™570) and anti-GSDMC2/3 (magenta; Opal^™690)on mouse colon.
Panel B : anti-SARM1 staining myenteric nerve plexus in mouse colon.
Panel C : anti-MUC2 staining goblet cells in mouse colon.
Panel D : anti-GSDMC2/3 staining epithelium in mouse colon.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab309195, ab272692 and ab229896 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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.

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

• WB

Lab

Western blot - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

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

Negative control : spleen (PMID : 17350798).

Exposure time : 54 seconds.

All lanes:

Western blot - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] (<a href='/en-us/products/primary-antibodies/gsdmc2-gsdmc3-antibody-epr20890-48-ab229896'>ab229896</a>) at 1/1000 dilution

Lane 1:

Mouse stomach tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 54 kDa

false

• WB

Lab

Western blot - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

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

The image was kindly provided by Dr. Jiahuai Han (Xiamen University, China).

Exposure time : 60 seconds.

All lanes:

Western blot - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] (<a href='/en-us/products/primary-antibodies/gsdmc2-gsdmc3-antibody-epr20890-48-ab229896'>ab229896</a>) at 1/1000 dilution

Lane 1:

Wild-type mouse esophagus tissue lysate at 20 µg

Lane 2:

mGSDMC1-4 knockout mouse esophagus tissue lysate at 20 µg

Lane 3:

Wild-type mouse stomach tissue lysate at 20 µg

Lane 4:

mGSDMC1-4 knockout mouse stomach tissue lysate at 20 µg

Lane 5:

Wild-type mouse small intestine tissue lysate at 20 µg

Lane 6:

mGSDMC1-4 knockout mouse small intestine tissue lysate at 20 µg

Lane 7:

Wild-type mouse colon tissue lysate at 20 µg

Lane 8:

mGSDMC1-4 knockout mouse colon tissue lysate at 20 µg

Lane 9:

Wild-type mouse caecum tissue lysate at 20 µg

Lane 10:

mGSDMC1-4 knockout mouse caecum 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 1/100000 dilution

Observed band size: 56 kDa

false

• WB

Lab

Western blot - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] - BSA and Azide free (AB269859)

This data was developed using ab229896, the same antibody clone in a different buffer formulation.

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

The image was kindly provided by Dr. Jiahuai Han (Xiamen University, China).

Exposure time : 60 seconds.

All lanes:

Western blot - Anti-GSDMC2 + GSDMC3 antibody [EPR20890-48] (<a href='/en-us/products/primary-antibodies/gsdmc2-gsdmc3-antibody-epr20890-48-ab229896'>ab229896</a>) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), whole cell lysate at 20 µg

Lane 2:

HEK-293T transfected with mGSDMC1 expression vector, whole cell lysate at 20 µg

Lane 3:

HEK-293T transfected with mGSDMC2 expression vector, whole cell lysate at 20 µg

Lane 4:

HEK-293T transfected with mGSDMC3 expression vector, whole cell lysate at 20 µg

Lane 5:

HEK-293T transfected with mGSDMC4 expression vector, whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 56 kDa

false