Anti-Calpain 2 antibody [EPR29833-94]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded (A) PC-3 (human prostate adenocarcinoma epithelial cell) cell pellet (B) 293T (human embryonic kidney epithelial cell) cell pellet (C) LNCaP (human prostate carcinoma epithelial cell) cell pellet (D) Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cell pellet tissue labeling Calpain 2 with ab325140 at 1/2000 (0.259 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining (A) PC-3 cell pellet and (B) 293T cell pellet, weak staining on (C) LNCaP cell pellet, no staining on (D) Jurkat cell pellet.
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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling Calpain 2 with ab325140 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung carcinoma.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Calpain 2 with ab325140 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling Calpain 2 with ab325140 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human stomach.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Calpain 2 with ab325140 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded Mouse lung cancer tissue labeling Calpain 2 with ab325140 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung cancer.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Calpain 2 with ab325140 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Supplier Data

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

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

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

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (ab325140) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 1 µg

Lane 2:

PC-3 transfected with siRNA specifically targeting CAPN2 whole cell lysate at 1 µg

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

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

Low expression : liver

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

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (ab325140) at 1/1000 dilution

Lane 1:

Human stomach tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human liver cancer tissue lysate at 20 µg

Secondary

All lanes:

Western blot at 1/2000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

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

Low expression : LNCaP, Jurkat

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

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (ab325140) at 1/1000 dilution

Lane 1:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

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

Secondary

All lanes:

Western blot at 1/100000 dilution

Observed band size: 76 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

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

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

Exposure time : Lanes1-2 : 26 seconds, lanes3-4 : 1 second, lane 5 : 26 seconds

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (ab325140) at 1/1000 dilution

Lane 1:

C2C12 (mouse myoblast) whole cell lysate at 20 µg

Lane 2:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 3:

Mouse stomach tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Rat 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: 76 kDa,36 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Calpain 2 with ab325140 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Weak staining on human liver.
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• WB

Supplier Data

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (AB325140)

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

This antibody does not cross-react with recombinant human CAPN1, CAPN3, CAPN8, CAPN9 or CAPN11 by western blot.

In Western blot, Anti-His tag antibody (ab15149) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Calpain 2 antibody [EPR29833-94] (ab325140) at 1/1000 dilution

Lane 1:

His-tagged human CAPN2 fragment at 10 ng

Lane 2:

His-tagged human CAPN1 fragment at 10 ng

Lane 3:

His-tagged human CAPN3 fragment at 100 ng

Lane 4:

His-tagged human CAPN8 fragment at 10 ng

Lane 5:

His-tagged human CAPN9 fragment at 10 ng

Lane 6:

His-tagged human CAPN11 fragment 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: 34 kDa

false

Exposure time: 26s