Invitrogen

TIMP2 Monoclonal Antibody (F27 P3 A4)

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

4 References

View all (52) TIMP2 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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TIMP2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of TIMP2 was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with TIMP2 (F27 P3 A4) Mouse Monoclonal Antibody (Product # MA1-774) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

TIMP2 Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse pancreas tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature and probed with a TIMP2 monoclonal antibody (Product # MA1-774) at a dilution of 1:100 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

TIMP2 Antibody in Western Blot (WB) — Western blot was performed using Anti-TIMP2 Monoclonal Antibody (F27 P3 A4) (Product # MA1-774) and a 24kDa band corresponding to TIMP2 was observed in HT-1080 but was absent in Daudi which is reported to be negative. Whole cell extracts (30 µg lysate) of HT-1080 (Lane 1), HT-1080 treated with Protein Transport Inhibitor (PTI; 1x for 4 hours) (Lane 2), Daudi (Lane 3) and Daudi treated with Protein Transport Inhibitor (1x for 4 hours) (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). An uncharacterized band (*) was observed at ~18kDa in the negative models.

TIMP2 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lysates tested owing to their inherent genetic constitution. Relative expression of TIMP2 was observed in HT-1080 as compared to Daudi using Anti-TIMP2 Monoclonal Antibody (F27 P3 A4) (Product # MA1-774) in Western Blot. {RE}

Product Details

MA1-774

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500	View 3 publications 3 publications
Immunohistochemistry (IHC)	-	View 1 publication 1 publication
Immunohistochemistry (Paraffin) (IHC (P))	1:50-1:500	-
Immunocytochemistry (ICC/IF)	1-2 µg/mL	-

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human, Mouse, Plant
Host/Isotype	Mouse / IgG2a, kappa
Class	Monoclonal
Type	Antibody
Clone	F27 P3 A4
Immunogen	Synthetic peptide corresponding to residues D(56) S G N D I Y G N P I(66) of mouse TIMP-2. View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein A
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_889483

Product Specific Information

MA1-774 contains 100 µg of in vitro produced, protein A purified antibody (1 mg/mL) in PBS containing 1 mg/mL BSA and 0.05% sodium azide.

MA1-774 detects TIMP-2 from mouse samples.

MA1-774 has been successfully used in Western blot, IF and immunohistochemistry procedures. By Western blot, MA1-774 detects a ~18-24 kDa band representing TIMP-2 from mouse platelets. It is recommended to block with 3% BSA.

The MA1-774 immunogen is a synthetic peptide corresponding to residues D(56) S G N D I Y G N P I(66) of mouse TIMP-2. This sequence is 100% conserved in human, rat, rabbit, chicken, and bovine TIMP-2

Target Information

TIMP-1 and TIMP-2 are inhibitory enzymes of matrix metalloproteinase family, and are thought to be of great importance in the maintenance of connective tissue integrity. TIMP-2 shows the highest binding affinity to both the latent (pro) and active forms of 72 kDa Type IV collagenase (also known as MMP-2 or gelatinase A). It also has affinity for the active form of 92 kDa Type IV collagenase (also known as MMP-9 or gelatinase B). TIMP-2 inhibits the proteolytic invasiveness of tumor cells and normal placental trophoblast cells.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Protective Effects of Kuding Tea (Ilex kudingcha C. J. Tseng) Polyphenols on UVB-Induced Skin Aging in SKH1 Hairless Mice.

Molecules (Basel, Switzerland)

Yi R,Zhang J,Sun P,Qian Y,Zhao X

MA1-774 was used in Western Blotting to study the protective effects of Kuding tea polyphenols on ultraviolet B-induced skin injury of SKH1 hairless mice.

Wed Mar 13 00:00:00 EDT 2019

A Genetic Screen Identifies PRP18a, a Putative Second Step Splicing Factor Important for Alternative Splicing and a Normal Phenotype in Arabidopsis thaliana.

G3 (Bethesda, Md.)

Kanno T,Lin WD,Chang CL,Matzke M,Matzke AJM

MA1-774 was used in Western Blotting to identify a putative second step splicing factor, important for alternative splicing and a normal phenotype in Arabidopsis thaliana.

Wed Mar 28 00:00:00 EDT 2018

CHD4 Has Oncogenic Functions in Initiating and Maintaining Epigenetic Suppression of Multiple Tumor Suppressor Genes.

Cancer cell

Xia L,Huang W,Bellani M,Seidman MM,Wu K,Fan D,Nie Y,Cai Y,Zhang YW,Yu LR,Li H,Zahnow CA,Xie W,Chiu Yen RW,Rassool FV,Baylin SB

MA1-774 was used in Immunohistochemistry to identify that CHD4 knockdown activates silenced tumour suppressor genes, revealing their role for blunting colorectal cancer cell proliferation, invasion, and metastases.

Mon May 08 00:00:00 EDT 2017

Inhibition of Smooth Muscle β-Catenin Hinders Neointima Formation After Vascular Injury.

Arteriosclerosis, thrombosis, and vascular biology

Riascos-Bernal DF,Chinnasamy P,Gross JN,Almonte V,Egaña-Gorroño L,Parikh D,Jayakumar S,Guo L,Sibinga NES

MA1774 was used in western blot to determine the impact of smooth muscle cell beta-catenin to vascular homeostasis and arterial injury

Mon May 01 00:00:00 EDT 2017

Bioinformatics

Protein Aliases: CSC-21K; Metalloproteinase inhibitor 2; testicular secretory protein Li 57; TIMP 2; TIMP-2; Tissue inhibitor of metalloproteinases 2

Gene Aliases: CSC-21K; D11Bwg1104e; DDC8; Timp-2; TIMP2

UniProt ID: (Human) P16035

Entrez Gene ID: (Human) 7077, (Mouse) 21858

Molecular Function: protease inhibitor