Anti-MMP9 antibody [EP1255Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1255Y] (AB137867)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lymphoma tissue labelling MMP9 with unpurified ab137867 at 1/100.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1255Y] (AB137867)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling MMP9 with unpurified ab137867 at 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1255Y] (AB137867)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling MMP9 with purified ab137867 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Lab

Western blot - Anti-MMP9 antibody [EP1255Y] (AB137867)

Blocking buffer and concentration 5% NFDM/TBST
Diluting buffer and concentration 5% NFDM/TBST

The expression of MMP-9 can be stimulated by various agents, such as inflammatory cytokine, growth factor, and 12-O-tetradecanoylphorbol-13-acetate (TPA) (PMID : 21047770, 28969043).

Treatment increasing the expression of MMP-9 is recommended when using this antibody.

All lanes:

Western blot - Anti-MMP9 antibody [EP1255Y] (ab137867) at 1/1000 dilution

Lane 1:

LoVo (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

A549 (Human lung carcinoma epithelial cell) serum starved overnight whole cell lysate at 20 µg

Lane 7:

A549 (Human lung carcinoma epithelial cell) serum starved overnight, then treated with 80nM TPA for 24 hours whole cell lysate at 20 µg

Lane 8:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) serum starved overnight whole cell lysate at 20 µg

Lane 9:

MDA-MB-231 (Human breast adenocarcinoma epithelial cell) serum starved overnight, then treated with 200nM TPA for 24 hours whole cell lysate at 20 µg

Lane 10:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell 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: 78 kDa

Observed band size: 84 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-MMP9 antibody [EP1255Y] (AB137867)

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Western blot - Anti-MMP9 antibody [EP1255Y] (ab137867) at 1/1000 dilution

All lanes:

U937 cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 15 kDa,78 kDa

Observed band size: 78 kDa

false

• WB

Lab

Western blot - Anti-MMP9 antibody [EP1255Y] (AB137867)

Western blot : Rabbit Monoclonal[EP1255Y] to MMP9 ab137867 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 78 kDa in Wild-type A549 TPA-treated (80nM, 24h) cell lysates with no signal observed at this size in Wild-type A549 Vehicle Control TPA (0nM, 24h) cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-MMP9 antibody [EP1255Y] (ab137867) at 1/1000 dilution

Lane 1:

Wild-type A549 TPA-treated (80nM, 24h) at 20 µg

Lane 2:

Wild-type A549 Vehicle Control TPA (0nM, 24h) at 20 µg

Lanes 3 - 4:

Western blot - Human MMP9 knockout A549 cell line (ab286527) at 20 µg

Lane 5:

Human Lung at 20 µg

Lane 6:

MOLT-4 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 78 kDa

Observed band size: 78 kDa

false