重组Anti-TIMP1抗体[EPR1550] (ab109125)

False colour image of Western blot: Anti-TIMP1 antibody [EPR1550] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109125 was shown to bind specifically to TIMP1. A band was observed at 30 kDa in wild-type HeLa cell lysates with no signal observed at this size in TIMP1 knockout cell line ab264022 (knockout cell lysate ab260091). The identity of bands observed at higher molecular weights has not been determined. To generate this image, wild-type and TIMP1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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.

Lanes 1-4: Merged signal (red and green). Green - ab109125 observed at 26 kDa. Red - loading control ab7291 observed at 50 kDa.

ab109125 Anti-TIMP1 antibody [EPR1550] was shown to specifically react with TIMP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261740 (knockout cell lysate ab257291) was used. Wild-type and TIMP1 knockout samples were subjected to SDS-PAGE. ab109125 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue sections labeling TIMP1 with purified ab109125 at 1:8000 dilution (0.21 μg/ml). Heat mediated antigen retrieval was performed using EDTA buffer, pH 9.0. Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling TIMP1 with purified ab109125 at 1:8000 dilution (0.21 μg/ml). Heat mediated antigen retrieval was performed using EDTA buffer, pH 9.0. Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Unpurified ab109125, at 1/250 dilution staining TIMP1 in paraffin-embedded human liver tissue, by Immunohistochemistry.

Blocking and diluting buffer: 5% NFDM/TBST