Anti-TIMP1 antibody [EPR18352]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] (AB211926)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human lung cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TIMP1 antibody [EPR18352] (AB211926)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (Human colorectal adenocarcinoma cell line) cells labeling TIMP1 with ab211926 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HT-29 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab211926 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] (AB211926)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human colon neuroendocrine cell is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] (AB211926)

Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human prostate cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] (AB211926)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human islet is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TIMP1 antibody [EPR18352] (AB211926)

Immunohistochemical analysis of paraffin-embedded human medullary thyroid carcinoma tissue labeling TIMP1 with ab211926 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human medullary thyroid carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TIMP1 antibody [EPR18352] (AB211926)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SK-OV-3 (Human ovarian cancer cell line) cells labeling TIMP1 with ab211926 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SK-OV-3 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab211926 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR18352] (AB211926)

False colour image of Western blot : Anti-TIMP1 antibody [EPR18352] 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, ab211926 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 [EPR18352] (ab211926) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lanes 1 - 5:

Western blot - Human TIMP1 knockout HeLa cell line (ab264022)

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

Secondary

Lanes 1 - 5:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 5:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 30 kDa

false

• WB

Supplier Data

Western blot - Anti-TIMP1 antibody [EPR18352] (AB211926)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16517973).

All lanes:

Western blot - Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution

All lanes:

Human prostate cancer lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 26 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-TIMP1 antibody [EPR18352] (AB211926)

Blocking/Dilution buffer : 5% NFDM/TBST.

Binding in these cell lines was extremely weak and requires optimization.

All lanes:

Western blot - Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution

Lane 1:

HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 2:

SK-OV-3 (Human ovarian cancer cell line) whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 26 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR18352] (AB211926)

False colour image of Western blot : Anti-TIMP1 antibody [EPR18352] 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, ab211926 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 [EPR18352] (ab211926) 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

false

• WB

Lab

Western blot - Anti-TIMP1 antibody [EPR18352] (AB211926)

False colour image of Western blot : Anti-TIMP1 antibody [EPR18352] staining at 1/1000 dilution, shown in black. In Western blot, ab211926 was shown to bind specifically to TIMP1. 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 before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) at 1/50000 dilution and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution

Lane 1:

A549 Vehicle Control BFA (0 u/mL, 6 h) cell lysate at 20 µg

Lane 2:

A549 Treated BFA (5 u/mL, 6 h) cell lysate at 20 µg

Predicted band size: 23 kDa

Observed band size: 30 kDa

false