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Human MMP-2(Matrix Metalloproteinase 2) ELISA Kit

Cat:E-EL-H1445 Citations (19)
Manual MSDS
Synonyms:

MMP2、CLG4、CLG4A、MMP-II、MONA、TBE-1、Gelatinase A

Price: $ 495

Price: $ 396

Price: $ 150

Price: Inquire

Price: Inquire

Size:
96T 48T 24T 96T*10 96T*5
Quantity:
  • Reactivity: Human
  • Detection Range: 0.78-50 ng/mL
  • Sensitivity: 0.47 ng/mL
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Product Details
Test Principle This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human MMP-2. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MMP-2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MMP-2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human MMP-2. You can calculate the concentration of Human MMP-2 in the samples by comparing the OD of the samples to the standard curve.
Assay Type Sandwich-ELISA
size 96T / 48T / 24T / 96T*10 / 96T*5
Assay Time 3.5h
Detection Method Colormetric
Detection Range 0.78-50 ng/mL
Sensitivity 0.47 ng/mL
Sample Volume 100μL
Sample Type serum, plasma and other biological fluids
Specificity This kit recognizes Human MMP-2 in samples. No significant cross-reactivity or interference between Human MMP-2 and analogues was observed
Precision Both intra-CV and inter-CV are < 10%.
Recovery 80%-120%
Introduction This ELISA kit applies to the in vitro quantitative determination of Human MMP-2 concentrations in serum, plasma and other biological fluids.
Applications ELISA
Storage 2-8℃/-20℃
Database SwissProt:  P08253
Syonoyms MMP2,CLG4,CLG4A,MMP-II,MONA,TBE-1,Gelatinase A
Research Area Cancer Cell Biology Cardiovascular Metabolism
  • Q1:Can ELISA measure the cell supernatant MMP-2 and MMP-9? I read the papers written by others, in addition to measuring the expression of these two indicators, but also to measure the activity, can ELISA measure the activity? What's the difference with gelatin enzymology?

    These two kits can be used for the detection of your cell supernatant. However, ELISA detection of cell supernatants is affected by factors such as media components, cell growth, and external drug stimulation conditions, and it is recommended that you conduct a preliminary experiment before the formal experiment. Gelatin enzyme spectrometry uses the reversible combination of SDS and MMPs in the sample to separate MMP-2 and MMP-9 in the sample, and then restores the activity of both through the bivalent metal ion buffer system. This test method is specifically aimed at the activity detection of MMP-2 and MMP-9. ELISA detects the content (that is, concentration) of the tested substance through the binding reaction of antigen and antibody, that is, captures the MMP-2 and MMP-9 of the sample through the specific antibodies of MMP-2 and MMP-9. The experimental principles of the two are different, and the activity of MMP-2 and MMP-9 cannot be detected by ELISA.

  • Q2:How to operate the "repeated freeze-thaw" process mentioned in tissue or cell sample processing?

    Tissue sample: After finishing tissue grinding, place it at -80℃ for 1h/ liquid nitrogen for 0.5h, and then gently shake it in a water bath at 30℃ to melt it quickly. Repeat this operation 1-2 times. Cell sample: Repeat the above freeze-thaw operation 2-3 times. If it is a membrane protein, it can be appropriately sonicated, but the temperature and frequency of ultrasound need to be controlled. It is recommended to add protease inhibitors to the sample in advance. PMSF(Cat.E-EL-SR002) is generally recommended.

  • Q3:Can the standard curve in the manual be used directly for calculation?

    The standard curve on the manual is an indicator curve, mainly used to show the shape of the standard curve after fitting and the quality control range of the highest/lowest OD value, you can not directly use. In addition, due to the influence of experimental operation, experimental environment, instrument parameter setting and other factors, the OD value of your actual standard curve may not be completely consistent with the instructions (overall high or low). In this case, it is recommended to control the first 4 blue wells in the color development stage with obvious color gradient and the maximum OD value of the standard curve above 1.2. If the correlation coefficient of the standard tune is above 0.99, it can be used as an effective standard curve.

  • Q4:The sample/reagent volume is not enough, can I reduce the sample volume uniformly?

    Neither can be reduced. The sample volume of each step should be strictly in accordance with the instructions, otherwise the experimental system will be changed, and the results will be inaccurate. If the customer really does not have enough sample volume, you can consult the technology to confirm whether the sample can be diluted, and judge the sample concentration through pre-experiment Degree situation. If the volume of the reagent is not enough, the number of holes to be tested can only be reduced according to the actual volume, and the detection can not be diluted.

  • Q5:How long was the ELISA test?

    Traditional Competion Method: about 2.0 h; Traditiona three-steps Sandwich method: about 3.5h; QuicKey ELISA: about 2.5h; QuicKey Pro ELISA: about 1.5h.

  • Q6:Does ELISA require technical replication? How to set the positive or negative control?

    It is recommended that customers do technical repetition, on the one hand can confirm the operation method, on the other hand can verify and improve the accuracy of the experimental results. A positive control in an ELISA test can be considered the standard product. Negative control is a blank matrix or a matrix solution with known low concentration that has homology and homogeneity with the object to be picked up, which is generally difficult to obtain, and can be set as a negative control if available. The routine is to set a blank control, that is, a sample diluent.

  • Q7:Why should a product with two item numbers of a target test different sample types?

    Different sample types of matrix are different, in order to reduce the influence of the matrix itself and improve the accuracy of routine sample measurement, usually set different systems of ELISA kits, and display with different product numbers. In addition, the concentration of some indicators varies greatly between different samples, so different products are used to meet customers' needs for ease of operation.

  • Q8:How can I tell if my sample needs to be diluted or processed?

    The detection range of the kit is not the same as the concentration range of the substance to be measured in the sample. It is recommended to estimate the concentration of the substance to be measured in the sample through relevant literature before the experiment and determine the actual concentration of the sample through pre-experiment. If it is a routine sample type, you can consult ELISA technical support to obtain the recommended dilution of the sample and schedule pre-laboratory testing. If the concentration of the substance to be measured in the sample is too high or too low, dilute or concentrate the sample appropriately. If the concentration of analyte in the sample is much lower than the minimum detection line of the kit, it is recommended to choose a kit with higher sensitivity for detection.

  • Q9:Is it necessary to set up multiple Wells for ELISA?

    In order to ensure the accuracy of the experimental results, we suggest that both standards and samples should be tested in duplicate. Of course, ELISA experiments do not necessarily have to be repeated /3 repeated Wells, customers can set the repeat or not according to their own needs.

  • Q10:How to perform ultrasonic processing?

    Using ultrasonic cell crusher, ultrasonic treatment of the suspension to lyse the cells (reference: Soniprep150 ultrasonic generator, ultrasonic treatment of 30s at an amplitude of 14μm, cell fragmentation; Or use an ultrasonic crusher, 200W, 2s/ times, gap 3s, total time 3-5min or 400 amp, 5s/ times, gap 10s, repeated 3-5 times), and then centrifuge at 2 to 8℃ at 1500×g for 10min, remove the cell debris, collect the supernatant. Attention should be paid to temperature during ultrasound.