Anti-Kallikrein-11 antibody [EPR31011-525R]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HT-29 (human colorectal adenocarcinoma epithelial cell) DU 145 (human prostate carcinoma epithelial cell) cells labelling Kallikrein-11 with ab326566 at 1/200 (2.49 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in HT-29 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Low expression : DU 145 (PMID : 10969073)

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326566 at 1/200 dilution, followed by ab150120 at 1/1000 dilution.-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

Immunohistochemical analysis of paraffin-embedded (A) : HT-29 (human colorectal adenocarcinoma epithelial cell) (B) : DU 145 (human prostate carcinoma epithelial cell) tissue labeling Kallikrein-11 with ab326566 at 1/500 (0.996 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) : HT-29, and no signal in (B) : DU 145.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling Kallikrein-11 with ab326566 at 1/500 (0.996 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human prostatic hyperplasia.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling Kallikrein-11 with ab326566 at 1/500 (0.996 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human endometrial cancer.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

Immunohistochemical analysis of paraffin-embedded Human salivary gland tissue labeling Kallikrein-11 with ab326566 at 1/500 (0.996 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human salivary gland.

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

Lab

Western blot - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

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

This antibody does not cross-react with full-length human Kallikrein-6, Kallikrein-13, Kallikrein-2 and Kallikrein-8.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(30-37KDa).

All lanes:

Western blot - Anti-Kallikrein-11 antibody [EPR31011-525R] (ab326566) at 1/1000 dilution

Lane 1:

His-tagged full-length human Kallikrein-11 recombinant protein at 10 ng

Lane 2:

His-tagged full-length human Kallikrein-6 recombinant protein at 10 ng

Lane 3:

His-tagged full-length human Kallikrein-13 recombinant protein at 10 ng

Lane 4:

His-tagged full-length human Kallikrein-2 recombinant protein at 10 ng

Lane 5:

His-tagged full-length human Kallikrein-8 recombinant protein at 10 ng

Secondary

All lanes:

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

Observed band size: 37 kDa,30-37 kDa

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

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

Low expression : DU 145 (PMID : 10969073), THP-1.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16467084).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Kallikrein-11 antibody [EPR31011-525R] (ab326566) at 1/1000 dilution

Lane 1:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

THP-1 (human monocytic leukemia monocyte) whole cell 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: 37 kDa,36 kDa

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

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

Low expression : kidney (PMID : 10969073).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16467084).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Kallikrein-11 antibody [EPR31011-525R] (ab326566) at 1/1000 dilution

Lane 1:

Human salivary gland tissue lysate at 20 µg

Lane 2:

Human stomach tissue lysate at 20 µg

Lane 3:

Human kidney 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: 37 kDa,36 kDa

false

Exposure time: 70s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kallikrein-11 antibody [EPR31011-525R] (AB326566)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Kallikrein-11 with ab326566 at 1/500 (0.996 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining in human kidney (PMID : 10969073).

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