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. View Details

4 References

View all (48) TIMP2 antibodies

Datasheet

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Datasheet

Questions & Answers

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TIMP2 Antibody in Immunofluorescence (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 Human 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 1:500 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 in Relative expression — 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

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

1-2 µg/mL

Immunofluorescence (IF)

1-2 µg/mL

Immunohistochemistry (Paraffin) (IHC (P))

1:50-1:500

Western Blot (WB)

1:500

View 3 publications 3 publications

Immunohistochemistry (IHC)

View 1 publication 1 publication

Product Specifications

Species

Human, Mouse

Published species

Human, Mouse, Plant

Expression System

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.

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.

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

Function(s)