Anti-MMP3 antibody [EP1186Y]

• WB

Lab

Western blot - Anti-MMP3 antibody [EP1186Y] (AB52915)

Western blot : Anti-MMP3 antibody [EP1186Y] (ab52915) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52915 was shown to bind specifically to MMP3. A band was observed at 55-60 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in MMP3 knockout cell line. To generate this image, wild-type and MMP3 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-MMP3 antibody [EP1186Y] (ab52915) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

MMP3 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Wild-type HeLa Treated: IL-1beta (10 ng/mL, 48 h) cell lysate at 20 µg

Lane 4:

Wild-type HeLa Vehicle control: IL-1beta (0 ng/mL, 48 h) cell lysate at 20 µg

Lane 5:

HeLa cell lysate at 20 µg

Lane 6:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 55 kDa,60 kDa

false

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MMP3 antibody [EP1186Y] (AB52915)

Immunofluorescence staining of HT-29 cells with purified ab52915 at a working dilution of 1/100, 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 100% methanol 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 ab52915 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP3 antibody [EP1186Y] (AB52915)

Immunohistochemical staining of paraffin embedded human liver with purified ab52915 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP3 antibody [EP1186Y] (AB52915)

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling MMP3 with ab52915 at a concentration of 0.01 μg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. The heat-mediated antigen retrieval was conducted with DISCOVERY cell conditioning solution (CC1) at 100 °C, pH 8.5 for 32 mins. ab52915 Anti-MMP3 antibody [EP1186Y] was incubated for 16 mins, at 37 °C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control. Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP3 antibody [EP1186Y] (AB52915)

Immunohistochemical staining of paraffin embedded rat kidney with purified ab52915 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP3 antibody [EP1186Y] (AB52915)

Immunohistochemical staining of paraffin embedded mouse liver with purified ab52915 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• WB

Lab

Western blot - Anti-MMP3 antibody [EP1186Y] (AB52915)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MMP3 antibody [EP1186Y] (ab52915) at 0.37 µg/mL

Lane 1:

Raji (Human Burkitt's lymphoma B lymphocyte )whole cell lysate at 20 µg

Lane 2:

HEK293 (Human embryonic kidney epithelial cell ) whole cell lysate at 20 µg

Lane 3:

Mouse lung lysate at 20 µg

Lane 4:

Mouse placenta 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

Predicted band size: 53 kDa

Observed band size: 50 kDa

false

Exposure time: 84s

• WB

Lab

Western blot - Anti-MMP3 antibody [EP1186Y] (AB52915)

Blocking and diluting buffer : 5% NFDM/TBST.

Exposure time : Lane 1 & 2 : 10 seconds
Lane 3 : 3 minutes

All lanes:

Western blot - Anti-MMP3 antibody [EP1186Y] (ab52915) at 0.37 µg/mL

Lane 1:

Rat brain lysate at 20 µg

Lane 2:

Rat liver lysate at 20 µg

Lane 3:

Rat spleen 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

Predicted band size: 53 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-MMP3 antibody [EP1186Y] (AB52915)

Blocking and diluting buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 3 minutes Lane 2 & 3 : 4 seconds

All lanes:

Western blot - Anti-MMP3 antibody [EP1186Y] (ab52915) at 0.37 µg/mL

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Mouse kidney lysate at 20 µg

Lane 3:

Mouse liver 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

Predicted band size: 53 kDa

false

• WB

Lab

Western blot - Anti-MMP3 antibody [EP1186Y] (AB52915)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-MMP3 antibody [EP1186Y] (ab52915) at 0.37 µg/mL

All lanes:

Rat kidney 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

Predicted band size: 53 kDa

Observed band size: 50 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-MMP3 antibody [EP1186Y] (AB52915)

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

All lanes:

Western blot - Anti-MMP3 antibody [EP1186Y] (ab52915) at 1/20000 dilution

All lanes:

MMP3 recombinant protein

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 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 50 kDa

false