Anti-MMP2 antibody [EPR1184] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

Immunofluorescence staining of PC-3 cells with purified ab92536 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab92536 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab92536 at a dilution of 1 in 400 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

Intracellular Flow Cytometry analysis of PC-3 (human prostate adenocarcinoma) cells labeling MMP2 with purified ab92536 at 1/180 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536).

• WB

Supplier Data

Western blot - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536).

The 72 KDa band is pro-MMP2 and the 69 KDa band is active-MMP2 reported by PMID 11489818 and 22190701.

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

This antibody shows low affinity in detecting mouse liver and HepG2 lysates which are positive for MMP2 reported by PMID 24096707 and 24297510.

All lanes:

Western blot - Anti-MMP2 antibody [EPR1184] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr1184-ab92536'>ab92536</a>) at 1/1000 dilution

Lane 1:

L6 (Rat skeletal muscle myoblast) whole cell lysates prepared in 1%SDS Hot lysis method at 20 µg

Lane 2:

Mouse liver lysates prepared in RIPA lysis method at 20 µg

Lane 3:

Mouse liver lysates prepared in 1%SDS Hot lysis method at 20 µg

Lane 4:

Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates prepared in RIPA lysis method at 20 µg

Lane 5:

Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates prepared in 1%SDS Hot lysis method at 20 µg

Lane 6:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method at 20 µg

Lane 7:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method 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

Predicted band size: 73 kDa

Observed band size: 69 kDa,72 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536). Blocking and diluting buffer and concentration : 5% NFDM /TBST. ab181602 was used as GAPDH loading control. Compared with ab92536, ab181286 has higher sensitivity. We recommend ab181286 as an alternative for testing MMP2 in western blot.

Lanes 1 - 5:

Western blot - Anti-MMP2 antibody [EPR1184] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr1184-ab92536'>ab92536</a>) at 1/1000 dilution

Lanes 1 - 5:

Western blot - Anti-MMP2 antibody [EPR17003-25] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr17003-25-ab181286'>ab181286</a>)

Lane 1:

HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Untreated HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HepG2 (Human hepatocellular carcinoma epithelial cell) treated with 300ng/ml BFA for 24 hours whole cell lysate at 20 µg

Lane 4:

Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell) 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

Predicted band size: 73 kDa

false

Exposure time: 60s

• WB

Lab

Western blot - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536). Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as a GAPDH loading control. Compared with ab92536, ab181286 has higher sensitivity. We recommend ab181286 as an alternative for testing MMP2 in western blot.

Lanes 1 - 3:

Western blot - Anti-MMP2 antibody [EPR1184] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr1184-ab92536'>ab92536</a>) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-MMP2 antibody [EPR17003-25] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr17003-25-ab181286'>ab181286</a>)

Lane 1:

Human plasma tissue lysate at 20 µg

Lane 2:

Human brain tissue lysate at 20 µg

Lane 3:

Human breast tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 73 kDa

Observed band size: 69 kDa,72 kDa

false

Exposure time: 60s

• WB

Unknown

Western blot - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536).

All lanes:

Western blot - Anti-MMP2 antibody [EPR1184] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr1184-ab92536'>ab92536</a>) at 1/1000 dilution

Lane 1:

L6 cell lysate at 10 µg

Lane 2:

Fetal heart lysate at 10 µg

Lane 3:

NIH/3T3 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 73 kDa

false

• WB

Lab

Western blot - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536).

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

72kDa : propeptide; 64kDa : active form

All lanes:

Western blot - Anti-MMP2 antibody [EPR1184] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr1184-ab92536'>ab92536</a>) at 1/5000 dilution

All lanes:

human skin at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 73 kDa

Observed band size: 64 kDa,72 kDa

false

• WB

Lab

Western blot - Anti-MMP2 antibody [EPR1184] - BSA and Azide free (AB271866)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92536).

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

72kDa : propeptide; 64kDa : active form

All lanes:

Western blot - Anti-MMP2 antibody [EPR1184] (<a href='/en-us/products/primary-antibodies/mmp2-antibody-epr1184-ab92536'>ab92536</a>) at 1/10000 dilution

Lane 1:

L6 cell lysate at 20 µg

Lane 2:

NIH/3T3 cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 73 kDa

Observed band size: 64 kDa,72 kDa

false