Anti-TIMP1 antibody [EPR1550]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR1550] (AB109125)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR1550] (AB109125)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR1550] (AB109125)

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

• WB

Unknown

Western blot - Anti-TIMP1 antibody [EPR1550] (AB109125)

All lanes:

Western blot - Anti-TIMP1 antibody [EPR1550] (ab109125) at 1/1000 dilution

Lane 1:

HL60+TPA cell lysates at 10 µg

Lane 2:

PBMC cell lysates at 10 µg

Predicted band size: 23 kDa

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• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR1550] (AB109125)

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.

All lanes:

Western blot - Anti-TIMP1 antibody [EPR1550] (ab109125) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TIMP1 knockout HeLa cell lysate at 20 µg

Lane 3:

HT-1080 treated with 200ng/ml 12-O-Tetradecanoylphorbol-13-acetate (TPA) for 24 hours, cell lysate at 20 µg

Lane 4:

Untreated HT-1080 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 26 kDa

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• WB

Lab

Western blot - Anti-TIMP1 antibody [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.

All lanes:

Western blot - Anti-TIMP1 antibody [EPR1550] (ab109125) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TIMP1 knockout HeLa cell lysate at 20 µg

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Lane 5:

U-87 MG cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 30 kDa

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• WB

Unknown

Western blot - Anti-TIMP1 antibody [EPR1550] (AB109125)

All lanes:

Western blot - Anti-TIMP1 antibody [EPR1550] (ab109125) at 1/1000 dilution

All lanes:

TIMP1 recombinant protein at 10 µg

Predicted band size: 23 kDa

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• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR1550] (AB109125)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-TIMP1 antibody [EPR1550] (ab109125) at 1/1000 dilution

Lane 1:

Human prostate lysates at 20 µg

Lane 2:

HT-1080 (Human fibrosarcoma epithelial cell) treated with 200ng/ml TPA for 24 hours whole cell lysates at 15 µg

Lane 3:

Untreated with HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 23 kDa

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