Knockout Tested Rabbit Recombinant Monoclonal TIMP1 antibody. Suitable for WB, ICC/IF, IHC-P and reacts with Human samples. Cited in 36 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
WB | ICC/IF | IHC-P | |
---|---|---|---|
Human | Tested | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Metalloproteinase inhibitor that functions by forming one to one complexes with target metalloproteinases, such as collagenases, and irreversibly inactivates them by binding to their catalytic zinc cofactor. Acts on MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13 and MMP16. Does not act on MMP14. Also functions as a growth factor that regulates cell differentiation, migration and cell death and activates cellular signaling cascades via CD63 and ITGB1. Plays a role in integrin signaling. Mediates erythropoiesis in vitro; but, unlike IL3, it is species-specific, stimulating the growth and differentiation of only human and murine erythroid progenitors.
CLGI, TIMP, TIMP1, Metalloproteinase inhibitor 1, Erythroid-potentiating activity, Fibroblast collagenase inhibitor, Tissue inhibitor of metalloproteinases 1, EPA, Collagenase inhibitor, TIMP-1
Knockout Tested Rabbit Recombinant Monoclonal TIMP1 antibody. Suitable for WB, ICC/IF, IHC-P and reacts with Human samples. Cited in 36 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Tissue Inhibitor of Metalloproteinase 1 (TIMP1) is a protein with a molecular mass of approximately 28 kDa. The protein inhibits metalloproteinases by binding to them preventing the breakdown of the extracellular matrix. This action regulates extracellular environment and tissue remodeling. TIMP1 is also known by the name Erythroid-Potentiating Activity (EPA). It is expressed in various tissues including the liver and the brain and can be found in high levels in the blood serum.
TIMP1 plays several essential roles beyond inhibiting metalloproteinases. It supports cell proliferation and influences apoptosis. It is part of a larger TIMP family and does not function as part of a complex per se but interacts closely with matrix metalloproteinases (MMPs). By regulating MMP activity TIMP1 balances processes like tissue growth and inflammatory responses which are important for normal development and repair.
TIMP1 participates in the regulation of the matrix metalloproteinase pathway impacting tissue homeostasis and repair. It closely associates with proteins such as MMP-9 and MMP-2 within this pathway. TIMP1 also plays a role in cytokine signaling pathways which influence immune responses by interacting with other signaling molecules and receptors that govern these pathways.
TIMP1's regulation of MMPs links it to cancer and cardiovascular disease progression. Overexpression of TIMP1 associates with tumor growth and metastasis where it can modulate interactions with other proteins like MMP-9 leading to altered tissue invasion and angiogenesis. Additionally TIMP1 imbalance connects to tissue fibrosis in cardiovascular disorders working in tandem with proteins such as MMP-2 which affects the structural remodeling and function of tissues within the cardiovascular system.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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False colour image of Western blot: Anti-TIMP1 antibody [EPR18352] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control 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
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 23 kDa
Observed band size: 26 kDa
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) (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) (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.
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
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
Exposure time: 5s
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) (Goat Anti-Rabbit IgG H&L (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 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 23 kDa
Observed band size: 26 kDa
Exposure time: 3min
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) (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) (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.
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) (Goat Anti-Rabbit IgG H&L (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 (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
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) (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) (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.
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) (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) (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.
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) (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) (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.
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 Anti-PKC delta (phospho S299) antibody [EPNCI119] 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
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 30 kDa
False colour image of Western blot: Anti-TIMP1 antibody [EPR18352] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control 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
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
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 30 kDa
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