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Human TIMP-3

Human TIMP-3 DuoSet ELISA Summary

Assay Type

Solid Phase   Sandwich ELISA

Format

96-well strip   plate

Assay Length

4.5 hours

Sample Type   & Volume Required Per Well

100 μL

Sensitivity

0.051 ng/mL

Assay Range

62.5 - 4,000   pg/mL (Cell Culture Supernates, Serum, Heparin Plasma, Saliva)

Specificity

Natural and   recombinant human TIMP-3

 

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

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human TIMP-3. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

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-3

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 3

Entrez Gene   IDs:

7078 (Human);   21859 (Mouse); 25358 (Rat)

Alternate   Names:

HSMRK222;   K222; K222TA2; metalloproteinase inhibitor 3; MIG-5 protein; Protein MIG-5;   pseudoinflammatory); SFD; TIMP metallopeptidase inhibitor 3; TIMP3; TIMP-3;   tissue inhibitor of metalloproteinase 3 (Sorsby fundus dystrophy; Tissue   inhibitor of metalloproteinases 3

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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