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Human TIMP-2

Human TIMP-2 Quantikine ELISA Kit Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

4.5   hours

Sample   Type & Volume Required Per Well

Cell   Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin   Plasma (10 uL), Saliva (10 uL), Urine (25 uL)

Sensitivity

0.064   ng/mL

Assay   Range

0.2 -   10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma,   Saliva, Urine)

Specificity

Natural   and recombinant human TIMP-2. This assay detects approximately 50% of   recombinant human TIMP-2 when complexed with recombinant human active MMP-9   in a 1:1 molar ratio

 

Cross-reactivity

<   0.5% cross-reactivity observed with available related molecules.< 50%   cross-species reactivity observed with species tested

Interference

No   significant interference observed with available related molecules.

Product Summary

The Quantikine Human TIMP-2 Immunoassay is a 4.5 hour solid phase ELISA designed to measure TIMP-2 levels in cell culture supernates, serum, plasma, and saliva. It contains CHO cell-expressed recombinant human TIMP-2, and antibodies raised against the recombinant protein. Natural human TIMP-2 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human TIMP-2.

Preparation and Storage

Shipping

The   product is shipped at ambient temperature. Upon receipt, store it immediately   at the temperature recommended below.

Storage

Store   the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TIMP-2

TIMPs-1 through -4 regulate the activity of zinc metalloproteases known as MMPs, ADAMs and ADAMTSs. Structurally, TIMPs contain two domains. The N-terminal domain binds to the active site of mature metalloproteases via a 1:1 non-covalent interaction, blocking access of substrates to the catalytic site. In addition, The C-terminal domain of TIMP-1 and TIMP-2 binds to the hemopexin- like domain of pro-MMP-9 and pro-MMP-2, respectively. The latter binding is essential for the cell surface activation of MMP-2 by MMP-14.

Long   Name:

Tissue   Inhibitors of Metalloproteinases 2

Entrez   Gene IDs:

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

Alternate   Names:

CSC-21Ktissue   inhibitor of metalloproteinase 2; metalloproteinase inhibitor 2; TIMP   metallopeptidase inhibitor 2; TIMP2; TIMP-2; Tissue inhibitor of   metalloproteinases 2

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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