Anti-ITGA11 antibody [EPR28689-21] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ITGA11 antibody [EPR28689-21] - BSA and Azide free (AB316250)

This data was developed using ab316249, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling ITGA11 with ab316249 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in tumor stroma interphase on Human breast carcinoma (PMID : 31605508).
The section was incubated with ab316249 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 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-ITGA11 antibody [EPR28689-21] - BSA and Azide free (AB316250)

This data was developed using ab316249, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cervical squamous carcinoma tissue labeling ITGA11 with ab316249 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in tumor stroma interphase on human cervical squamous carcinoma (PMID : 36003785).
The section was incubated with ab316249 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 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-ITGA11 antibody [EPR28689-21] - BSA and Azide free (AB316250)

This data was developed using ab316249, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling ITGA11 with ab316249 at 1/2000 (0.255 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human spleen.
The section was incubated with ab316249 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-ITGA11 antibody [EPR28689-21] - BSA and Azide free (AB316250)

This data was developed using ab316249, the same antibody clone in a different buffer formulation.

Negative control : 293T (PMID : 31605508).

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

All lanes:

Western blot - Anti-ITGA11 antibody [EPR28689-21] (<a href='/en-us/products/primary-antibodies/itga11-antibody-epr28689-21-ab316249'>ab316249</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell fresh lysate at 20 µg with 5% NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell fresh lysate at 20 µg with 5% NFDM/TBST

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: 160 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ITGA11 antibody [EPR28689-21] - BSA and Azide free (AB316250)

This data was developed using ab316249, the same antibody clone in a different buffer formulation.

Western blot : Anti-ITGA11 antibody (ab316249) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5]

(ab8245) loading control staining at 1/20000 dilution, shown in red.

In Western blot, ab316249 was shown to bind specifically to ITGA11. Target of interest was observed at 160 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in ITGA11 knockout cell line (lane 2, knockout cell line ab265494 / knockout cell lysate ab258929). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

Negative control : A431 (PMID : 31605508).

All lanes:

Western blot - Anti-ITGA11 antibody [EPR28689-21] (<a href='/en-us/products/primary-antibodies/itga11-antibody-epr28689-21-ab316249'>ab316249</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 1/2 volume of Odyssey Blocking Buffer (TBS)+ 1/2 volume of TBS

Lane 2:

ITGA11 knockout HeLa whole cell lysate at 20 µg with 1/2 volume of Odyssey Blocking Buffer (TBS)+ 1/2 volume of TBS

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg with 1/2 volume of Odyssey Blocking Buffer (TBS)+ 1/2 volume of TBS

Lane 4:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg with 1/2 volume of Odyssey Blocking Buffer (TBS)+ 1/2 volume of TBS

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution

Observed band size: 160 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-ITGA11 antibody [EPR28689-21] - BSA and Azide free (AB316250)

This data was developed using ab316249, the same antibody clone in a different buffer formulation.

Negative control : 4T1 (PMID : 26990302), C2C12 (PMID : 11518510).

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

All lanes:

Western blot - Anti-ITGA11 antibody [EPR28689-21] (<a href='/en-us/products/primary-antibodies/itga11-antibody-epr28689-21-ab316249'>ab316249</a>) at 1/1000 dilution

Lane 1:

MEF (mouse embryo fibroblast) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg with 5% NFDM/TBST

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: 160 kDa,36 kDa

false

Exposure time: 37s