5-NT Mouse 5-Nuclease ELISA Detection Kit

The products supplied by our company are for scientific research purposes only. The following are the product attributes:
Product full name:5-NT Mouse 5-Nuclease ELISA Detection Kit

English name: 5-NT ELISA Kit

Item number: BKE6322
Specification: 48T/96T

Service: We can provide free testing services, as well as product manuals and technical guidance

Storage environment: 2-8 ℃ low temperature, away from light, moisture-proof

Main ingredients: enzyme-linked immunosorbent assay (ELISA) plates, reagents, standards, etc.

Purpose of testing: Used to determine samples such as serum, plasma, and related fluids. For example, suitable for detecting specimens including serum, plasma, urine, pleural and peritoneal fluid, lavage fluid, cerebrospinal fluid, cell culture supernatant, tissue homogenate, etc Needed but not provided.

Reagent preparation:

1. Enzyme linked plate: one piece (96 well or 48 well).

2. Standard: 2 bottles (freeze-dried).

3. Sample Diluent: 1 × 20ml/bottle.

4. Biotin antibody diluent: 1 × 10ml/bottle.

5. HRP avidin diluent labeled with horseradish peroxidase: 1 × 10ml/bottle.

6. Biotin antibody labeling: 1 × 120 μ l/bottle (1:100)

7. Horseradish peroxidase labeled avidin (HRP avidin): 1 × 120 μ l/bottle (1:100)

8. TMB Substrate: 1 × 10ml/bottle.

9. Wash Buffer: 1 × 20ml/bottle, diluted 25 times with distilled water per bottle during use.

10. Stop Solution: 1 × 10ml/bottle (2N H2SO4).
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Composition and reagent preparation:

1. Enzyme linked plate: one piece (96 well)
2. Standard (freeze-dried): 2 bottles, dilute each bottle with sample diluent to 1ml before use, cover and let it stand for more than 10 minutes, then repeatedly invert/rub to aid dissolution, with a concentration of 100 ng/ml. After serial dilutions (note: do not directly dilute in the plate), dilute each bottle to 100 ng/ml ng/ml，50 ng/ml，25 ng/ml，12.5 ng/ml，6.25 ng/ml，3.12 ng/ml，1.56 ng/ml， The sample diluent is directly used as the standard concentration of 0 ng/ml and prepared within 15 minutes before use.
To prepare a 50 ng/ml standard: Take 0.5ml (not less than 0.5ml) of the above standard at 100 ng/ml and add it to an Eppendorf tube containing 0.5ml of sample diluent. Mix well, and repeat the same process for other concentrations.
3. Sample diluent: 1 × 20ml/bottle.
4. Test diluent A: 1 × 10ml/bottle.
5. Test diluent B: 1 × 10ml/bottle.
6. Test solution A: 1 × 120ul/bottle (1:100). Before use, dilute with test diluent A at a ratio of 1:100. Before dilution, prepare according to the pre calculated total amount required for each experiment (100ul/well). In actual preparation, an additional 0.1-0.2ml should be prepared. For example, a ratio of 10ul of test solution A to 990ul of test diluent A should be prepared, gently mixed, and prepared within one hour before use.
7. Dilute the detection solution B at a ratio of 1:100 with a dilution of 1:100 before use. The dilution method is the same as the detection solution A.
8. Substrate solution: 1 × 10ml/bottle.
9. Concentrated detergent: 1 × 30ml/bottle, diluted 25 times with distilled water per bottle during use.
10. Termination solution: 1 × 10ml/bottle (2N H2SO4).

Lactose Fermentation Medium (Lactose Fermentation Medium) 1000 (g)

Deoxycholic acid salt agar (DC) 250 (g)

Deoxycholic acid salt agar (DC) 1000 (g)

Hershey's medium 250 (g)

Hershey's bacterial enrichment solution 250 (g)

Hemolytic agar base 250 (g)

Gentian Purple Blood Agar Base 250 (g)

Strain culture medium 250 (g)

Tetracycline detection agar 250 (g)

MR-VP medium 250 (g)

Cha's culture medium 250 (g)

Crystal Violet Neutral Red Bile Salt Agar VRBA 250 (g)

Crystal Purple Neutral Red Bile Salt Agar 1000 (g)

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Operation steps:

1. Dilution of standard samples: This kit provides one original standard sample, and users can dilute it in a small test tube according to the following chart.

2. Sample addition: Set up blank wells (blank control wells do not contain samples or enzyme-linked immunosorbent assay reagents, all other steps are the same), standard wells, and wells for the test sample. Accurately add 50 μ l of the standard sample on the enzyme-linked immunosorbent assay (ELISA) coated plate, first add 40 μ l of sample diluent to the well of the test sample, and then add 10 μ l of the test sample (the final dilution of the sample is 5 times). Add the sample to the bottom of the enzyme-linked immunosorbent assay plate well, avoiding touching the well wall as much as possible, and gently shake and mix well.

3. Incubation: Cover the plate with a sealing film and incubate at 37 ℃ for 30 minutes.

4. Liquid preparation: Dilute 30 times the concentrated washing solution with distilled water and set aside for later use.

5. Washing: Carefully remove the sealing film, discard the liquid, shake dry, fill each hole with washing solution, let it stand for 30 seconds, then discard. Repeat this process 5 times and pat dry.

6. Enzyme addition: Add 50 μ l of enzyme labeled reagent to each well, except for blank wells.

7. Incubation: Follow the same procedure as in 3.

8. Washing: Follow the same procedure as in 5.

9. Color development: Add 50 μ l of color developing agent A50 to each well, then add 50 μ l of color developing agent B50, gently shake and mix, and develop color at 37 ℃ in the dark for 15 minutes.

10. Termination: Add 50 μ l of termination solution to each well to terminate the reaction (at this point, blue will turn yellow).

11. Measurement: Measure the absorbance (OD value) of each well in sequence using blank air conditioner zero and 450nm wavelength. The measurement should be conducted within 15 minutes after adding the termination solution.