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Product Info Summary
| SKU: | EK1302 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Rat |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Rat TIMP-2 ELISA Kit PicoKine®
SKU/Catalog Number
EK1302
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Rat TIMP-2 ELISA Kit PicoKine™ (96 Tests). Quantitate Rat Timp2 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.
Storage & Handling
Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
Cite This Product
Rat TIMP-2 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1302)
Clonality of Antibodies
See Datasheet for details
Immunogen
Expression system for standard: NS0; Immunogen sequence: C27-P220
Sensitivity
<10 pg/ml
Assay Range
156 pg/ml - 10,000 pg/ml
Standard Dilution Instructions
Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK1302 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK1302 is reactive to TIMP2 in Rat samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK1302 is guaranteed for ELISA in Rat by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of TIMP-2
TIMP-2 gen is encoded by 5 exons spanning 83 kb of genomic DNA. TIMP-2 is 83 kilobase pairs (kb) long with exon-intron splicing sites located in preserved positions among the three members of the TIMP family. The gene for tissue inhibitor of metalloproteinases-2 is localized on human chromosome arm 17q25. TIMP-2 abrogates angiogenic factor-induced endothelial cell proliferation in vitro and angiogenesis in vivo independent of MMP inhibition. The standard product used in this kit is recombinant human TIMP-2 with the molecular mass of 22Kda and 194 amino acid.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK1302-CAP | Anti-Rat TIMP2 Pre-coated 96-well strip microplate | 1 |
| EK1302-ST | Rat TIMP2 Standard | 2 vials, 10 ng/tube |
| EK1302-DA | Rat TIMP2 Biotinylated antibody (100x) | 100ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
| AR1106-1 | Sample Diluent | 30ml |
| AR1106-2 | Antibody Diluent | 12ml |
| AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
| AR1104 | Color Developing Reagent (TMB) | 10ml |
| AR1105 | Stop Solution | 10ml |
| AR1106-5 | Wash Buffer (25x) | 20ml |
| PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.028 | 0.056 | 0.118 | 0.205 | 0.402 | 0.849 | 1.345 | 1.970 |
Data Example Images
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Rat TIMP-2 PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect TIMP-2, Dilution ratio of 1:200, concentration in serum and plasma is 173-240 ng/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Rat TIMP-2 ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 240 | 1342 | 4533 | 227 | 1463 | 4769 |
| Standard deviation | 10.56 | 68.44 | 339.97 | 10.21 | 95.09 | 443.51 |
| CV (%) | 4.4% | 9.3% | 7.5% | 4.5% | 6.5% | 9.3% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of TIMP-2 in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 240 | 251 | 238 | 213 | 235 | 13.9 | 5.9% |
| Sample 2 | 1342 | 1286 | 1409 | 1375 | 1353 | 45.35 | 3.3% |
| Sample 3 | 4533 | 4407 | 4200 | 4479 | 4404 | 126.38 | 2.8% |
Protein Target Info & Infographic
Gene/Protein Information For TIMP2 (Source: Uniprot.Org, NCBI)
Gene Name
TIMP2
Full Name
Metalloproteinase inhibitor 2
Weight
24.399kDa
Superfamily
protease inhibitor I35 (TIMP) family
Alternative Names
CSC-21Ktissue inhibitor of metalloproteinase 2; metalloproteinase inhibitor 2; TIMP metallopeptidase inhibitor 2; TIMP2; TIMP-2; Tissue inhibitor of metalloproteinases 2 TIMP2 CSC-21K, DDC8 TIMP metallopeptidase inhibitor 2 metalloproteinase inhibitor 2|testicular secretory protein Li 57|tissue inhibitor of metalloproteinases 2
*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".For more info on TIMP2, check out the TIMP2 Infographic
We have 30,000+ of these available, one for each gene! check them out.
In this infographic you will see the following information for TIMP2: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].
Specific Publications For Rat TIMP-2 ELISA Kit PicoKine® (EK1302)
Hello CJ!
EK1302 has been cited in 7 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
High glucose stimulates cell proliferation and Collagen IV production in rat mesangial cells through inhibiting AMPK-KATP signaling
Zhang, B., Shi, Y.Q., Zou, J.J., Chen, X.F., Tang, W., Ye, F., & Liu, Z.M. (2017). High glucose stimulates cell proliferation and Collagen IV production in rat mesangial cells through inhibiting AMPK-KATP signaling. International Urology and Nephr...
Cavdar Z, Ozbal S, Celik A, Ergur Bu, Guneli E, Ural C, Camsari T, Guner Ga. Biotech Histochem. 2014 May;89(4):304-14. Doi: 10.3109/10520295.2013.847498. Epub 2013 Oct 28. The Effects Of Alpha-Lipoic Acid On Mmp-2 And Mmp-9 Activities In A Rat Ren...
Xu Xl, Ling Dy, Zhu Qy, Fan Wj, Zhang W. Eur J Pharmacol. 2013 Jan 5;698(1-3):370-8. Doi: 10.1016/J.Ejphar.2012.11.019. Epub 2012 Nov 21. The Effect Of 2,3,4',5-Tetrahydroxystilbene-2-0-??-D Glucoside On Neointima Formation In A Rat Artery Balloon...
Yu X, Zhu J, Mi M, Chen W, Pan Q, Wei M. Med Oncol. 2012 Mar;29(1):349-57. Doi: 10.1007/S12032-010-9770-2. Epub 2010 Dec 4. Anti-Angiogenic Genistein Inhibits Vegf-Induced Endothelial Cell Activation By Decreasing Ptk Activity And Mapk Activation.
Gishto A, Farrell K, Kothapalli Cr. J Biomed Mater Res A. 2015 Feb;103(2):693-708. Doi: 10.1002/Jbm.A.35217. Epub 2014 May 19. Tuning Composition And Architecture Of Biomimetic Scaffolds For Enhanced Matrix Synthesis By Murine Cardiomyocytes.
Kim Ks, Lee Ya, Choi Hm, Yoo Mc, Yang Hi. Rheumatol Int. 2012 Oct;32(10):3069-75. Epub 2011 Sep 13. Implication Of Mmp-9 And Urokinase Plasminogen Activator (Upa) In The Activation Of Pro-Matrix Metalloproteinase (Mmp)-13.
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Customer Reviews
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Customer Q&As
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8 Customer Q&As for Rat TIMP-2 ELISA Kit PicoKine®
Question
Q: What is the optimal O.D. value for TIMP-2 ELISA kit? I used your TIMP-2 ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain TIMP-2 even though the O.D. values are not very high?
Verified Customer
Verified customer
Asked: 2020-05-24
Answer
A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. an important assessment should be is whether your sample O.D. values are statistically significantly higher than your blank values. considering your assay, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in proportion to your positive controls. typically, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by converting cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this TIMP-2 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of TIMP-2 that can be declared/interpreted as positive by the above standard is 10pg/ml.
Boster Scientific Support
Answered: 2020-05-24
Question
Q: is it okay to use citrate plasma as samples in Rat TIMP-2 Picokine® ELISA Kit (Catalog # EK1302)?
G. Zhang
Verified customer
Asked: 2019-10-23
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can irreversibly bind metal ions from the functional domain of TIMP-2 causing degradation of its protein structure. TIMP-2 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the TIMP-2 ELISA, treating samples with various anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2019-10-23
Question
Why are there precipitates in the samples diluent buffer for EK1302?
Verified customer
Asked: 2019-05-29
Answer
For the Rat TIMP-2 ELISA Kit PicoKine™ (EK1302), the precipitates are fatty substances contained in BSA. It would not affect the performace of the kit.
Boster Scientific Support
Answered: 2019-05-30
Question
Do you have information regarding the target epitope for the antibody in your TIMP2 ELISA kit?
Verified customer
Asked: 2019-04-22
Answer
Unfortunately, we do not do epitope mapping for our antibodies, so we are only able to provide the immunogen sequence.
Boster Scientific Support
Answered: 2019-04-22
Question
Have you tested EK1302 on urine samples? In your opinion, it would work?
Verified customer
Asked: 2019-03-01
Answer
We have not validated Rat TIMP-2 ELISA Kit PicoKine™ EK1302 with urine samples. In theory, the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. For sample types that we have not tested for, we suggest your customer run pilot experiments to decide the optimal sample dilution.
Boster Scientific Support
Answered: 2019-03-01
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
D. Kumar
Verified customer
Asked: 2019-03-01
Answer
A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present in the product insert. Components in lysate and lysis buffer may impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2019-03-01
Question
Q: if the enzyme conjugated TIMP-2 antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the TIMP-2 antigen?
Verified Customer
Verified customer
Asked: 2019-02-28
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2019-02-28
Question
Q: we need your suggestion regarding the dilution ratio of serum samples for detection of TIMP-2 in Rat serum? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have low serum volumes so we like to dilute as much as possible.
Verified Customer
Verified customer
Asked: 2018-03-09
Answer
A: unable to understand the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the TIMP-2 ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.
Boster Scientific Support
Answered: 2018-03-09
