TIMP-2 Antibody

R&D Systems | Catalog # AF971

R&D Systems

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Neutralization, Dual RNAscope ISH-IHC Compatible

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunocytochemistry, Immunoprecipitation, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all TIMP-2 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human TIMP‑2
Cys27-Pro220
Accession # P16035

Specificity

Detects human and mouse TIMP‑2 in direct ELISAs and Western blots. In direct ELISAs, less than 2% cross‑reactivity with recombinant human TIMP-4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for TIMP-2 Antibody

Detection of Human TIMP-2 antibody by Western Blot.

Detection of Human TIMP‑2 by Western Blot.

Western blot shows Recombinant Human TIMP-2 Western Blot Standard Protein (Catalog # WBC023) and lysates of HeLa human cervical epithelial carcinoma cell line and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse TIMP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF971) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for TIMP-2 at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
TIMP‑2 antibody in Human Ovarian Cancer Tissue by Immunohistochemistry (IHC-P).

TIMP‑2 in Human Ovarian Cancer Tissue.

TIMP-2 was detected in immersion fixed paraffin-embedded sections of human ovarian cancer tissue using 15 µg/mL Goat Anti-Human/Mouse TIMP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF971) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counter-stained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western Blot Shows Human TIMP‑2 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and human TIMP-2 knockout HeLa human cervical epithelial carcinoma cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse TIMP‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF971) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for TIMP‑2 at approximately 22 kDa (as indicated) in the parental HeLa human cervical epithelial carcinoma cell line, but is not detectable in knockout HeLa human cervical epithelial carcinoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Neutralization of TIMP‑2 Activity by Human TIMP‑2 Antibody.

Neutralization of TIMP‑2 Activity by Human TIMP‑2 Antibody.

Recombinant Human MMP-2 (0.2 µg/mL, Catalog # 902-MPNor 902-MP) activity is measured in the presence of Recombinant Human/Mouse TIMP-2 (0.143 µg/mL, Catalog # 971-TM) that has been preincubated with increasing concentrations of Goat Anti-Human TIMP-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF971). The ND50 is typically 2.6 µg/mL.

Detection of TIMP‑2 in Human Placenta.

Formalin-fixed paraffin-embedded tissue sections of human placenta were probed for TIMP2 mRNA (ACD RNAScope Probe, catalog #470358; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using goat anti-human TIMP2 polyclonal antibody (R&D Systems catalog # AF971) at 5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to decidual cells.
Human TIMP-2 ELISA Standard Curve

Human TIMP-2 ELISA Standard Curve

Recombinant Human TIMP‑2 (Catalog # 971-TM) was serially diluted and captured by Mouse Anti-Human TIMP‑2 Monoclonal Antibody (Catalog # MAB9711) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human/Mouse TIMP‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF971) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Detection of TIMP-2 by Western Blot

Detection of TIMP-2 by Western Blot

Knockout (KO) of lnc-FANCI-2 in CaSki cells affects expression of cellular soluble receptors.(A) Dot blots show differentially expressed soluble receptors in delta Pr-A9 cells when compared to those in the wild-type (WT) CaSki cells. A Proteome Profiler Human sReceptor Array was used to examine 105 cellular soluble receptors. Label in red, upregulated receptors; label in blue, downregulated receptors. (B) Quantification of differentially expressed and/or released soluble receptors (A) in delta Pr-A9 cells when compared to those in the WT CaSki cells. Error bars represent standard deviation of replicates in each proteome array. The dashed line indicates the threshold of fold change (FC) with a score above or below the threshold (-/+0.3 FC) being determined as differentially expressed and/or released receptors. (C) Validation of several differentially expressed soluble receptors by immunoblot analysis using specific antibodies. Tubulin served as an internal loading control. *Nonspecific band in MCAM immunoblot.Figure 4—source data 1.Knockout (KO) of lnc-FANCI-2 in CaSki cells affects expression of cellular soluble receptors.Figure 4—source data 2.Knockout (KO) of lnc-FANCI-2 in CaSki cells affects expression of cellular soluble receptors.Knockout (KO) of lnc-FANCI-2 in CaSki cells affects expression of cellular soluble receptors.Knockout (KO) of lnc-FANCI-2 in CaSki cells affects expression of cellular soluble receptors. Image collected and cropped by CiteAb from the following open publication (https://elifesciences.org/articles/102681), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for TIMP-2 Antibody

Application
Recommended Usage

Dual RNAscope ISH-IHC Compatible

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human placenta

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human ovarian cancer tissue and normal human ovarian array

Knockout Validated

TIMP‑2 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in TIMP‑2 knockout HeLa human cervical epithelial carcinoma cell line.

Western Blot

1 µg/mL
Sample: Recombinant Human TIMP-2 Western Blot Standard Protein (Catalog # WBC023), HeLa human cervical epithelial carcinoma cell line and human placenta tissue

Neutralization

Measured by its ability to neutralize Recombinant Human TIMP-2 (0.143 µg/mL, Catalog # 971-TM) inhibition of Recombinant Human MMP‑2 (0.2 µg/mL, Catalog # 902-MPN or 902-MP) cleavage of the fluorogenic peptide substrate Mca‑PLGL‑Dpa‑AR‑NH2 (10 µM, Catalog # ES001). The Neutralization Dose (ND50) is typically 2.6 µg/mL.

Reviewed Applications

Read 2 reviews rated 4 using AF971 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: TIMP-2

Tissue inhibitors of metalloproteinases or TIMPs are a family of proteins that regulate the activation and proteolytic activity of the zinc enzymes known as matrix metalloproteinases (MMPs). There are four members of the family, TIMP-1, TIMP-2, TIMP-3, and TIMP-4. TIMP-2 is a non N-glycosylated protein with a molecular mass of 22 kDa produced by a wide range of cell types, which inhibits MMPs non-covalently by the formation of binary complexes. TIMP-2 also has erythroid-potentiating and cell growth promoting activities.

Long Name

Tissue Inhibitors of Metalloproteinases 2

Alternate Names

TIMP2

Entrez Gene IDs

7077 (Human); 21858 (Mouse); 29543 (Rat)

Gene Symbol

TIMP2

UniProt

P16035

Additional TIMP-2 Products

Product Documents for TIMP-2 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for TIMP-2 Antibody

For research use only

Citations for TIMP-2 Antibody

Customer Reviews for TIMP-2 Antibody (2)

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  • Human/Mouse TIMP-2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Adult brain
    Species: Mouse
    Verified Customer | Posted 02/16/2022
    Dilution 1:300
    TIMP-2 Antibody AF971
  • Human/Mouse TIMP-2 Antibody
    Name: Ruhul Choudhury
    Application: Immunohistochemistry
    Sample Tested: Human first trimester decidua
    Species: Goat
    Verified Customer | Posted 01/23/2017
    sodium citrate antigen retrieval
    TIMP-2 Antibody AF971

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