ab39184 binds to TIMP3. It recognizes the glycosylated and unglycosylated forms of TIMP3, and works against native or reduced TIMP3. Ab39184 does not cross react with the other TIMP family members (TIMP1, TIMP2, TIMP4).
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml.
1/1000 - 1/5000. Detects a band of approximately 24, 30 kDa (predicted molecular weight: 24 kDa). Dilution optimised using Chromogenic detection.
Use at an assay dependent concentration. PubMed: 20448060
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Complexes with metalloproteinases (such as collagenases) and irreversibly inactivates them by binding to their catalytic zinc cofactor. May form part of a tissue-specific acute response to remodeling stimuli. Known to act on MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MMP-13, MMP-14 and MMP-15.
Involvement in disease
Defects in TIMP3 are the cause of Sorsby fundus dystrophy (SFD) [MIM:136900]. SFD is a rare autosomal dominant macular disorder with an age of onset in the fourth decade. It is characterized by loss of central vision from subretinal neovascularization and atrophy of the ocular tissues. Generally, macular disciform degeneration develops in the patients eye within 6 months to 6 years.
Belongs to the protease inhibitor I35 (TIMP) family. Contains 1 NTR domain.
Secreted > extracellular space > extracellular matrix.
ICC/IF image of ab39184 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab39184, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Frozen sections) - Anti-TIMP3 antibody (ab39184)This image is courtesy of an Abreview submitted by Julia Watson
ab39184 staining TIMP3 in mouse fallopian tube tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with PBST (PBS / 0.1% v/v Triton X-100) and blocked with 3% donkey serum / 1% BSA in PBST for 1 hour at 25°C. Samples were incubated with primary antibody (1/200 in blocking buffer) for 48 hours at 25°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
Positive TIMP3 staining is seen in the extracellular space. The image shows DAPI counterstain, red field (autofluorescence), green field antibody staining and overlay.
Yun J et al. IL-32 gamma reduces lung tumor development through upregulation of TIMP-3 overexpression and hypomethylation. Cell Death Dis9:306 (2018).
Read more (PubMed: 29467412) »
Han M et al. Hispidulin inhibits hepatocellular carcinoma growth and metastasis through AMPK and ERK signaling mediated activation of PPAR?. Biomed Pharmacother103:272-283 (2018).
Read more (PubMed: 29656183) »