Anti-GSDMA antibody [EPR19858-17] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-GSDMA antibody [EPR19858-17] - BSA and Azide free (AB231043)

GSDMA was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate with ab214818 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214818 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1 : Hela cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate 10 μg (Input).
Lane 2 : ab209847 IP in HeLa cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab214818 in HeLa cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate (-).
Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : Less than 1 second.

The observed molecular mass is consistent with the literature (PMID : 17350798).

The observed molecular masses lower than 50 kDa are degraded expressed GSDMA protein. The cells were kindly provided by our collaborator Dr. Feng Shao, NIBS.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214818).

All lanes:

Immunoprecipitation - Anti-GSDMA antibody [EPR19858-17] (<a href='/en-us/products/primary-antibodies/gsdma-antibody-epr19858-17-ab214818'>ab214818</a>)

Predicted band size: 49 kDa

false

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GSDMA antibody [EPR19858-17] - BSA and Azide free (AB231043)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse stomach tissue labeling GSDMA with ab214818 at 1/50 dilution (green), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Positive staining in cells localized in the differentiated region of stomach epithelia of mouse stomach tissue section (PMID : 17471240; 19051310) is observed. Counter stained with DAPI (blue).
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1000 dilution.
Antigen retrieval is not needed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214818).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-GSDMA antibody [EPR19858-17] - BSA and Azide free (AB231043)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse breast skin tissue labeling GSDMA with ab214818 at 1/50 dilution (green), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1000 dilution. Positive staining in skin keratinocytes and hair follicle cells on mouse breast skin, negative staining in basal and stromal cells (PMID : 17471240; 19051310) is observed. Counter stained with DAPI (blue).
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1000 dilution.
Antigen retrieval is not needed.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214818).

• WB

Supplier Data

Western blot - Anti-GSDMA antibody [EPR19858-17] - BSA and Azide free (AB231043)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214818).

Blocking/Dilution buffer : 5%NFDM/TBS.

The observed molecular mass is consistent with the literature (PMID : 17350798).

The observed molecular masses lower than 50 kDa are degraded expressed GSDMA protein. The cells were kindly provided by our collaborator Dr. Feng Shao, NIBS.

All lanes:

Western blot - Anti-GSDMA antibody [EPR19858-17] (<a href='/en-us/products/primary-antibodies/gsdma-antibody-epr19858-17-ab214818'>ab214818</a>) at 1/1000 dilution

Lane 1:

HeLa cells, whole cell lysate at 20 µg

Lane 2:

HeLa cells stably transfected with human GSDMA expression vector, containing a DDDDK-tag, whole cell lysate lysate at 20 µg

Lane 3:

HeLa cells stably transfected with mouse GSDMA expression vector, containing a DDDDK-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

Predicted band size: 49 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-GSDMA antibody [EPR19858-17] - BSA and Azide free (AB231043)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214818).

Blocking/Dilution buffer : 5%NFDM/TBST.

Exposure times :

Lanes 1/3 : 102 seconds

Lane 2 : 3 minutes

This image is produced using super sensitivity ECL substrate. We strongly suggest the customer to use higher sensitivity ECL substrate when developing the blot.

All lanes:

Western blot - Anti-GSDMA antibody [EPR19858-17] (<a href='/en-us/products/primary-antibodies/gsdma-antibody-epr19858-17-ab214818'>ab214818</a>) at 1/1000 dilution

Lane 1:

Human stomach lysate at 10 µg

Lane 2:

Human skin lysate at 10 µg

Lane 3:

Rat skin lysate at 10 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: 49 kDa

false

• WB

Supplier Data

Western blot - Anti-GSDMA antibody [EPR19858-17] - BSA and Azide free (AB231043)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214818).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GSDMA antibody [EPR19858-17] (<a href='/en-us/products/primary-antibodies/gsdma-antibody-epr19858-17-ab214818'>ab214818</a>) at 1/1000 dilution

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

HeLa cells stably transfected with mouse GSDMA2 expression vector, containing a DDDDK-tag, whole cell lysate at 20 µg

Lane 3:

HeLa cells stably transfected with mouse GSDMA3 expression vector, containing a DDDDK-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 1/50000 dilution

Predicted band size: 49 kDa

false

Exposure time: 70s