Anti-MMP3 antibody [EP1186Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP3 antibody [EP1186Y] - BSA and Azide free (AB214794)

This IHC data was generated using the same anti-MMP3 antibody clone, EP1186Y, in a different buffer formulation (cat# 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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MMP3 antibody [EP1186Y] - BSA and Azide free (AB214794)

This ICC data was generated using the same anti-MMP3 antibody clone, EP1186Y, in a different buffer formulation (cat# 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] - BSA and Azide free (AB214794)

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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52915).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP3 antibody [EP1186Y] - BSA and Azide free (AB214794)

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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52915).

• WB

Lab

Western blot - Anti-MMP3 antibody [EP1186Y] - BSA and Azide free (AB214794)

This data was developed using the same antibody clone in a different buffer formulation (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] (<a href='/en-us/products/primary-antibodies/mmp3-antibody-ep1186y-ab52915'>ab52915</a>) 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