Anti-ADA antibody [EPR25429-117] (BSA and Azide free)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling ADA with ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection) was used. Positive staining on human stomach. The section was incubated with ab300050 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 (ab300050) 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling ADA with ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection) was used. Negative control : no staining on human cerebrum (PMID : 2606352). The section was incubated with ab300050 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 (ab300050) 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling ADA with ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection). Positive staining on the stromal cells of human liver. The section was incubated with ab300050 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 (ab300050) 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labelling ADA with ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection). Positive staining on human tonsil (PMID : 30778076) (PMID : 6360330). The section was incubated with ab300050 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 (ab300050) 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized Jurkat (human T cell leukemia T lymphocyte) cells lebelling ADA with ab300050 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and membranous staining in the Jurkat cell line.

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : PBS was used instead of primary antibody followed by secondary antibody (ab150081) Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human thymus tissue labelling ADA with ab300050 at 1/2000 followed by a ready to use LeicaDS9800 (BOND^™ Polymer Refine Detection). Positive staining on human thymus (PMID : 2606352). The section was incubated with ab300050 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 (ab300050) 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.

• IP

Supplier Data

Immunoprecipitation - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

ADA was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10 µg with ab300050 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab300050 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10 µg (Input)

Lane 2 : ab300050 IP in Jurkat whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300050 in Jurkat whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-ADA antibody [EPR25429-117] (<a href='/en-us/products/primary-antibodies/ada-antibody-epr25429-117-ab300050'>ab300050</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate (Input) at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/ada-antibody-epr25429-117-ab300050'>ab300050</a> IP in Jurkat whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/ada-antibody-epr25429-117-ab300050'>ab300050</a> in Jurkat whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 40 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

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

Exposure time :

Lane1-2 : 1 second;

Lane3-4 : 3.25 seconds.

Low expression : human liver(PMID : 2606352)

All lanes:

Western blot - Anti-ADA antibody [EPR25429-117] (<a href='/en-us/products/primary-antibodies/ada-antibody-epr25429-117-ab300050'>ab300050</a>) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 3:

Human stomach tissue lysate at 20 µg

Lane 4:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 40 kDa

Observed band size: 41 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-ADA antibody [EPR25429-117] (BSA and Azide free) (AB300051)

This data was developed using ab300050, the same antibody clone in a different buffer formulation.

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

Exposure time :

Lane1 : 3.25 second;

Lane2 : 37 seconds.

All lanes:

Western blot - Anti-ADA antibody [EPR25429-117] (<a href='/en-us/products/primary-antibodies/ada-antibody-epr25429-117-ab300050'>ab300050</a>) at 1/1000 dilution

Lane 1:

Human thymus tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 40 kDa

Observed band size: 41 kDa

false