Anti-Cathepsin L/MEP antibody [EPR30887-518]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling Cathepsin L/MEP with ab326362 at 1/4000 (0.133 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon 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 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Cathepsin L/MEP with ab326362 at 1/4000 (0.133 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive 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 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Cathepsin L/MEP with ab326362 at 1/4000 (0.133 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen.

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 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Cathepsin L/MEP with ab326362 at 1/4000 (0.133 µg/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 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling Cathepsin L/MEP with ab326362 at 1/4000 (0.133 µg/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 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Cathepsin L/MEP with ab326362 at 1/4000 (0.133 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kindey.

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 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded :
(A) Wild type HEK-293T (human epithelial cell line from embryonic kidney) cell pellet
(B) CTSL knockout HEK293T cell pellet
labeling Cathepsin L/MEP with ab326362 at 1/1000 (0.533 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Wild type HEK-293T cell pellet, no staining on (B) CTSL knockout HEK293T cell pellet

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 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling Cathepsin L/MEP with ab326362 at 1/4000 (0.133 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Weak 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

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

• WB

Lab

Western blot - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

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

In Western blot, ab326362 was shown to bind specifically to CTSL. Target of interest was observed at 25 kDa in wild-type HEK-293T cell lysates (lane 1) with no signal observed at this size in CTSL HEK-293T knockout cell lysate(ab257213)/cell line (lane 2)(ab255449).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) 1 : 100000 (15Kda).

All lanes:

Western blot - Anti-Cathepsin L/MEP antibody [EPR30887-518] (ab326362) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 80 µg

Lane 2:

Western blot - Human CTSL (Cathepsin L/MEP) knockout HEK-293T cell lysate (<a href='/ko/products/cell-lysates/human-ctsl-cathepsin-l-mep-knockout-hek-293t-cell-lysate-ab257213'>ab257213</a>) at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 25 kDa,15 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 11483509, PMID : 33483465).

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

All lanes:

Western blot - Anti-Cathepsin L/MEP antibody [EPR30887-518] (ab326362) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 4:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 5:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 6:

Human liver tissue lysate at 40 µg

Lane 7:

Human kidney tissue lysate at 40 µg

Lane 8:

Human spleen tissue lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 38 kDa,30 kDa,25 kDa,36 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-Cathepsin L/MEP antibody [EPR30887-518] (AB326362)

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

This antibody does not cross-react with human CTSK, CTSS or CTSV.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(30-75KDa).

All lanes:

Western blot - Anti-Cathepsin L/MEP antibody [EPR30887-518] (ab326362) at 1/1000 dilution

Lane 1:

No tagged human CTSL recombinant fragment at 10 ng

Lane 2:

His-tagged human CTSK recombinant fragment at 10 ng

Lane 3:

His-tagged human CTSS recombinant fragment at 10 ng

Lane 4:

His-tagged human CTSV recombinant fragment at 10 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 75 kDa,30-75 kDa

false

Exposure time: 48s