Name: Rat NACHT,LRRandPYDdomains containing protein 3 (NLRP3) ELISA Kit
Brand: Biological test kits, ELISA (enzyme-linked immunosorbent assay) kits, chemical reagents-Shanghai IDEAL MEDICAL TECH Co. Ltd.
SKU: ADL-EL-RT01063
Availability: InStock
Rating: 5 (1 reviews)

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Home > ELISA Kit > Researchuse ELISA kit > Rat ELISA kit

Rat NACHT,LRRandPYDdomains containing protein 3 (NLRP3) ELISA Kit

Description

Experimental principle:

Coat the target antibody in a 96 well microplate to form a solid phase carrier. Add standard or specimen to each well, where the target is bound to the antibody attached to the solid phase carrier. Then add the microbialized target antibody. Wash the unbound biotin antibody, add HRP label and avidin, wash thoroughly again, and add TMB substrate for color development. TMB is converted to blue under the catalysis of peroxidase and to the final yellow under the action of acid. The depth of color is positively correlated with the target in the sample. Measure the absorbance (O.D. value) at a wavelength of 450nm using an enzyme-linked immunosorbent assay (ELISA) reader and calculate the sample concentration.

Kit composition:

Name	96 determinations	48 determinations
Microelisa stripplate	12*8strips	12*4strips
Standard	0.3ml*6tubes	0.3ml*6tubes
Sample Diluent	6.0ml	3.0ml
HRP-Conjugate reagent	10.0ml	5.0ml
20X Wash solution	25ml	15ml
Chromogen Solution A	6.0ml	3.0ml
Chromogen Solution B	6.0ml	3.0ml
Stop Solution	6.0ml	3.0ml
Closure plate membrane	2	2
User manual	1	1
Sealed bags	1	1

Test kit operation steps:
1. Prepare all r e a g e n ts before starting assay procedure. It is recommended that

all Standards and Samples be added in duplicate to the Microelisa Stripplate.

2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to

standard well.

3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing

sample well; Blank well doesn’t add anyting.

4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip

and incubate for 60 minutes at 37°C.

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

washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle,

manifold dispenser or autowasher. Complete removal of liquid at each step is

essential to good performance. After the last wash, remove any remaining Wash

Solution by aspirating or decanting. Invert the plate and blot it against clean paper

towels.

6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well.

Gently mix and incubate for 15 minutes at 37°C. Protect from light.

7. Add 50μl Stop Solution to each well. The color in the wells should change

from blue to yellow. If the color in the wells is green or the color change does not

appear uniform, gently tap the plate to ensure thorough mixing.

8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader

within 15 minutes

Self provided experimental equipment required for the experiment:

1. ELISA reader (450nm)

2. High precision sampler and nozzle: 0.5-10uL, 2-20uL, 20-200uL, 200-1000uL

3. 37 ℃ constant temperature box

4. Distilled water or deionized water

Experimental result calculation:

Draw standard curve: In an Excel worksheet, use the standard concentration as the horizontal axis and the corresponding OD value as the vertical axis to draw a linear regression curve of the standard. Calculate the concentration values of each sample according to the curve equation.

Test kit performance: Please request detailed electronic instructions from customer service personnel!

Technical tip:

1.When mixing or redissolving protein solutions, try to avoid foaming as much as possible.

2.To avoid cross infection, it is necessary to replace the nozzle when configuring different concentration standards, loading samples, and adding different reagents. Additionally, please use different pipettes for different reagents.

3. During each incubation, please use the sealing glue correctly to ensure the accuracy of the results.

4. The mixed chromogenic substrate should be colorless before being loaded onto the plate. Please store it away from light; If the microplate changes from being searched to different depths of blue.

5. The order of terminating the liquid on the plate should be consistent with the order of the color developing substrate on the plate; After adding the termination solution, the color inside the hole changed from blue to yellow; If there is green inside the hole, it indicates that the liquid inside the hole is not mixed evenly; Please mix thoroughly.

Product expiration date: 6 months

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Rat NACHT,LRRandPYDdomains containing protein 3 (NLRP3) ELISA Kit

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