Antibodies, Proteins, Kits and Reagents for Life Science

351 792 ₸ Product size

100 µl 351 792 ₸

Order now and get it on Tuesday March 02, 2021

Anti-TIMP1 antibody [EPR18352] (ab211926)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18352] to TIMP1
Suitable for: IHC-P, WB, ICC/IF
Knockout validated
Reacts with: Human

Product name

Anti-TIMP1 antibody [EPR18352]
See all TIMP1 primary antibodies
Description

Rabbit monoclonal [EPR18352] to TIMP1
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human prostate cancer lysate; HeLa, SK-OV-3 and HT-29 whole cell lysates. IHC-P: Human colon, pancreas, lung cancer, medullary thyroid carcinoma and prostate cancer tissues. ICC/IF: SK-OV-3 and HT-29 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18352
Isotype

IgG
Research areas

Western blot - Anti-TIMP1 antibody [EPR18352] (ab211926)

All lanes : Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : TIMP1 knockout HeLa cell lysate
Lane 3 : HT-1080 treated with 200ng/ml 12-O Tetradecanoylphorbol-13-acetate (TPA) for 24 hours; cell lysate
Lane 4 : Untreated HT-1080 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 23 kDa
Observed band size: 26 kDa
why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab211926 observed at 26 kDa. Red - loading control ab7291 observed at 50 kDa.

ab211926 Anti-TIMP1 antibody [EPR18352] 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. ab211926 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) - 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.
Western blot - Anti-TIMP1 antibody [EPR18352] (ab211926)

Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution + Human prostate cancer lysate at 10 µg

Secondary
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 why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 16517973).
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.
Western blot - Anti-TIMP1 antibody [EPR18352] (ab211926)

All lanes : Anti-TIMP1 antibody [EPR18352] (ab211926) at 1/1000 dilution

Lane 1 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 2 : SK-OV-3 (Human ovarian cancer cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 23 kDa
Observed band size: 26 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

Binding in these cell lines was extremely weak and requires optimization.
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.
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.
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.
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.
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.
Anti-TIMP1 antibody [EPR18352] (ab211926)

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