Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822)

This data was developed using ab317821, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's lymphoma tissue labeling ATG4B with ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human non-Hodgkin's lymhoma.
The section was incubated with ab317821 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed 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-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822)

This data was developed using ab317821, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling ATG4B with ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The section was incubated with ab317821 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed 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-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822)

This data was developed using ab317821, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colorectal carcinoma tissue labeling ATG4B with ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colorectal carcinoma (PMID : 24991826).
The section was incubated with ab317821 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed 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-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822)

This data was developed using ab317821, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ATG4B with ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : very weak staining on human colon (PMID : 24991826).
The section was incubated with ab317821 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed 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-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822)

This data was developed using ab317821, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell pellet (B) ATG4B knockout HeLa cell pellet tissue labeling ATG4B with ab317821 at 1/500 (1.052 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Wild-type HeLa cell pellet, no staining on (B) ATG4B knockout HeLa cell pellet.
The section was incubated with ab317821 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822)

This data was developed using ab317821, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ATG4B antibody [EPR28756-91] (<a href='/en-us/products/primary-antibodies/atg4b-antibody-epr28756-91-ab317821'>ab317821</a>) at 1/1000 dilution

All lanes:

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

Observed band size: 44 kDa

true

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-ATG4B antibody [EPR28756-91] - BSA and Azide free (AB317822)

This data was developed using ab317821, the same antibody clone in a different buffer formulation.

In Western blot, ab317821 was shown to bind specifically to ATG4B. A band was observed at 44 kDa in wild-type Hela cell lysates whereas loss of signal was observed in the ATG4B knockout cell line (ab265814/knockout lysate ab257364).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 15 seconds, Lanes 3-5 : 48 seconds

All lanes:

Western blot - Anti-ATG4B antibody [EPR28756-91] (<a href='/en-us/products/primary-antibodies/atg4b-antibody-epr28756-91-ab317821'>ab317821</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

ATG4B knockout HeLa whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

Ramos (human Burkitts lymphoma B lymphocyte) 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 1/20000 dilution

Observed band size: 44 kDa,36 kDa

false