Anti-COL11A1 antibody [EPR28734-175]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COL11A1 antibody [EPR28734-175] (AB324519)

Immunohistochemical analysis of paraffin-embedded (A) A204 (human muscle rhabdomyosarcoma cell) cell pellet, (B) SAOS-2 (human osteosarcoma epithelial cell) cell pellet, (C) MCF7 (human breast adenocarcinoma epithelial cell) cell pellet and (D) A549 (human lung carcinoma epithelial cell) cell pellet labeling COL11A1 with ab324519 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) A204 and (B) SAOS-2 cell pellet, no staining on (C) MCF7 cell pellet, weak and scattered on (D) A549 cell pellet (PMID : 22322826, PMID : 25761876). The primary antibody was incubated 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-COL11A1 antibody [EPR28734-175] (AB324519)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling COL11A1 with ab324519 at 1/1000 (0.515 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human skeletal muscle. The primary antibody was incubated 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-COL11A1 antibody [EPR28734-175] (AB324519)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling COL11A1 with ab324519 at 1/1000 (0.515 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human kidney. The primary antibody was incubated 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-COL11A1 antibody [EPR28734-175] (AB324519)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human COL11A1 expression vector containing a his tag; (B) HEK-293T transfected with empty vector containing a his tag labeling COL11A1 with ab324519 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a human COL11A1 expression vector containing a his tag, no staining on (B) HEK-293T transfected with empty vector containing a his tag. The primary antibody was incubated 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-COL11A1 antibody [EPR28734-175] (AB324519)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling COL11A1 with ab324519 at 1/1000 (0.515 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on stroma in human breast carcinoma (PMID : 37760937). The primary antibody was incubated 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-COL11A1 antibody [EPR28734-175] (AB324519)

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

Low expression : MCF7, A549 (PMID : 27373316).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-COL11A1 antibody [EPR28734-175] (ab324519) at 1/1000 dilution

Lane 1:

A-204 (human muscle rhabdomyosarcoma cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial cell) 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: 80-280 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-COL11A1 antibody [EPR28734-175] (AB324519)

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-COL11A1 antibody [EPR28734-175] (ab324519) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human COBA1 expression vector containing a His-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

Observed band size: 80-280 kDa

false

Exposure time: 7s