Anti-MMP13 antibody [EPR25858-87] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MMP13 antibody [EPR25858-87] - BSA and Azide free (AB315268)

This data was developed using ab315267, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling MMP13 with ab315267 at 1/1000 (0.491 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing positive staining in 293T cells transfected with a mouse MMP13 expression vector containing a myc tag, and showing no staining in 293T cells transfected with a mouse MMP8/MMP10/empty expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP13 antibody [EPR25858-87] - BSA and Azide free (AB315268)

This data was developed using ab315267, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MMP13 with ab315267 at 1/100 (4.91 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skeletal muscle. The section was incubated with ab315267 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-MMP13 antibody [EPR25858-87] - BSA and Azide free (AB315268)

This data was developed using ab315267, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cartilage tissue labeling MMP13 with ab315267 at 1/100 (4.91 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cartilage (PMID : 24108368). The section was incubated with ab315267 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.

• IP

Supplier Data

Immunoprecipitation - Anti-MMP13 antibody [EPR25858-87] - BSA and Azide free (AB315268)

This data was developed using ab315267, the same antibody clone in a different buffer formulation.

MMP13 was immunoprecipitated from 0.35 mg Mouse articular cartilage tissue lysate with ab315267 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315267 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-MMP13 antibody [EPR25858-87] (<a href='/en-us/products/primary-antibodies/mmp13-antibody-epr25858-87-ab315267'>ab315267</a>) at 1/1000 dilution

Lanes 1 - 2:

Mouse articular cartilage tissue lysate

Lane 2:

<a href='/en-us/products/primary-antibodies/mmp13-antibody-epr25858-87-ab315267'>ab315267</a> at 1/30 IP in Mouse articular cartilage tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/mmp13-antibody-epr25858-87-ab315267'>ab315267</a> in mouse articular cartilage tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MMP13 antibody [EPR25858-87] - BSA and Azide free (AB315268)

This data was developed using ab315267, the same antibody clone in a different buffer formulation.

Negative control : brain, skeletal muscle (PMID : 8609233).

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

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

All lanes:

Western blot - Anti-MMP13 antibody [EPR25858-87] (<a href='/en-us/products/primary-antibodies/mmp13-antibody-epr25858-87-ab315267'>ab315267</a>) at 1/1000 dilution

Lane 1:

Mouse articular cartilage tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue 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: 60 kDa,48 kDa,36 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-MMP13 antibody [EPR25858-87] - BSA and Azide free (AB315268)

This data was developed using ab315267, the same antibody clone in a different buffer formulation.

This antibody does not cross-react with mouse MMP8 and MMP10.

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

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

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

All lanes:

Western blot - Anti-MMP13 antibody [EPR25858-87] (<a href='/en-us/products/primary-antibodies/mmp13-antibody-epr25858-87-ab315267'>ab315267</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 5 µg

Lane 2:

293T cells transfected with a mouse MMP13 expression vector containing a myc-His-tag®, whole cell lysate at 5 µg

Lane 3:

293T cells transfected with a mouse MMP8 expression vector containing a myc-His-tag®, whole cell lysate at 5 µg

Lane 4:

293T cells transfected with a mouse MMP10 expression vector containing a myc-His-tag®, whole cell lysate at 5 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: 48-60 kDa,36 kDa

false

Exposure time: 3s