Human cyclic guanosine phosphate (cGMP) Elisa kit
Instructions for use of human cyclic guanosine phosphate (cGMP) Elisa kit
The human cyclic guanosine phosphate (cGMP) Elisa kit is for research use only.
Detection range: 96T 1.5 ng / ml-48 ng / ml
Purpose: This kit is used to determine the content of cGMP in human serum, plasma and related liquid samples.
Experimental principle: This kit uses the double antibody sandwich method to determine the level of human cyclic guanosine phosphate (cGMP) in the specimen. The microplate was coated with purified human cyclic guanosine monophosphate (cGMP) antibody to make a solid-phase antibody, and cyclic guanosine monophosphate (cGMP) was added to the monoclonal antibody-coated microwells in turn, followed by HRP-labeled cyclic guanosine phosphate The glycoside (cGMP) antibody binds to form an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and into the final yellow under the action of acid. The color depth is positively correlated with cGMP in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450 nm, and the concentration of human cyclic guanosine phosphate (cGMP) in the sample was calculated by a standard curve.
Human cyclic guanosine phosphate (cGMP) Elisa kit consists of:
1 30 times concentrated washing solution 20ml × 1 bottle 7 stop solution 6ml × 1 bottle
2 Enzyme label reagent 6ml × 1 bottle 8 standard (96 ng / ml) 0.5ml × 1 bottle
3 Enzyme label coated plate 12 wells × 8 strips 9 standard dilutions 1.5ml × 1 bottle
4 Sample diluent 6ml × 1 bottle 10 instructions 1 copy
5 Developer A solution 6ml × 1 bottle 11 2 sealing film
6 Developer B liquid 6ml × 1 / bottle 12 1 sealed bag
Specimen requirements:
1. Samples containing NaN3 cannot be detected because NaN3 inhibits horseradish peroxidase (HRP) activity.
2. The specimens should be extracted as soon as possible after collection. The extraction should be carried out according to relevant literature. If the test cannot be performed immediately, the specimen can be stored at -20 ℃, but repeated freezing and thawing should be avoided
Steps:
1. Dilution of standard products: This kit provides one original standard product. The user can perform dilution in a small test tube according to the following chart.
48 ng / ml No. 5 standard 150μl primary standard added 150μl standard dilution
24 ng / ml No. 4 standard 150μl No. 5 standard added 150μl standard dilution
12 ng / ml No. 3 standard 150μl No. 4 standard added 150μl standard diluent
6 ng / ml No. 2 standard 150μl No. 3 standard added 150μl standard dilution
3 ng / ml No. 1 standard 150μl No. 2 standard added 150μl standard dilution
2. Adding samples: set up blank wells (the blank control wells do not add samples and enzyme reagents, the rest of the steps are the same), standard wells, sample wells to be tested. Accurately add 50μl of standard on the enzyme-coated plate, add 40μl of sample diluent to the sample well, and then add 10μl of sample to be tested (the final dilution of the sample is 5 times) Add the sample and add the sample to the bottom of the well of the microplate, try not to touch the wall of the well, shake gently to mix.
3. Incubation: seal the plate with a sealing film and incubate at 37 ° C for 30 minutes.
4. Mixing solution: Dilute 30 times concentrated washing liquid with distilled water 30 times and reserve for use
5. Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with the washing liquid, let it stand for 30 seconds and then discard, repeat this 5 times and pat dry.
6. Add enzyme: add 50μl of enzyme label reagent to each well, except blank well.
7. Incubation: The operation is the same as 3.
8. Washing: The operation is the same as 5.
9. Color development: add 50μl of developer A to each well, and then add 50μl of developer B, mix gently, and develop at 37 ° C in the dark for 15 minutes.
10. Termination: Add 50 μl of stop solution to each well to stop the reaction (at this time, the blue color turns to yellow).
11. Determination: Measure the absorbance (OD value) of each well in sequence with the blank air conditioner at zero and 450 nm wavelength. The measurement should be carried out within 15 minutes after adding the stop solution.
Calculation: take the concentration of the standard as the abscissa and the OD as the ordinate, draw a standard curve on the coordinate paper, and find the corresponding concentration from the standard curve according to the OD value of the sample; multiply by the dilution factor; or use the standard The linear regression equation of the standard curve is calculated by the concentration and OD value of the sample, the OD value of the sample is substituted into the equation, the sample concentration is calculated, and then multiplied by the dilution factor, which is the actual concentration of the sample.
Precautions:
1. The kit should be taken out of the refrigerated environment and equilibrated at room temperature for 15-30 minutes before use. If the enzyme-coated plate is unsealed after opening, the strip should be stored in a sealed bag.
2. Crystals may be precipitated in the concentrated washing liquid, which can be heated and dissolved in the water bath during dilution, and the results will not be affected during washing.
3. The sample adder should be used in each step of sample addition, and its accuracy should be regularly checked to avoid test errors. It is best to control the sampling time within 5 minutes. If there are many specimens, it is recommended to use a volley gun to add samples.
4. Please make a standard curve at the same time of each measurement, it is best to make a double hole. If the content of the substance to be tested in the specimen is too high (the OD value of the sample is greater than the OD value of the first well of the standard well), please dilute it with a certain multiple of the sample diluent (n times) before measuring, and finally multiply the total dilution when calculating Multiple (× n × 5).
5. The sealing film is limited to one-time use to avoid cross contamination.
6. Please keep the substrate away from light.
7. Strictly follow the instructions, and the test results must be determined based on the reading of the microplate reader.
8. All samples, washing liquid and various wastes should be treated as infectious agents.
9. The components of different batches of this reagent shall not be mixed.
10. If there is any difference with the English manual, the English manual shall prevail.
Storage conditions and validity period:
1. Kit storage: 2-8 ℃.
2. Validity: 6 months
Dog
Products
There are over six hundred million dogs in the world today. Dogs have been a big part of human life for thousands of years.
Many dog owners see their dogs as their own family and they are always looking for great Dog Products to keep their furry friends happy and healthy.
As a supplier of Dog Products, we supply over thousands of great items for your dog's daily life, for their enjoyment, and in turn, your enjoyment as well.
For fun and
entertainment, we have a huge range of Dog Toys. Including nylon
chew toys, rubber dog toys, vinyl toys, latex toys, plastic dog toys, and
plush toys. Dog toys are a great way for dogs to enjoy themselves, either alone,
or with their human friends. Some dog toys are designed for the purpose of
dental hygiene and most well formulated well designed toys promote good
chewing habits.
For sleeping or
resting, we provide outdoor wooden or plastic dog houses. If your dog is an
indoor type, we provide soft foam and cotton dog beds to keep them comfortable
and warm. Stainless steel and metal wire dog cages are also available.
For walking your
dog and for basic safety, we have a huge range of collar, leads, harnesses,
safety chains and leather or plastic muzzles.
We have a very
large range of food bowls, water bowls and water drinkers. Many designs and
colours of plastic bowls, classic ceramic bowls and long lasting stainless steel
bowls. We also have portable drinkers and foldable silicone bowls for
travelling with your dog, or for use when taking your dogs for walks.
Nowadays, dog
owners like to have their dogs with them whenever possible. Holidays, shopping,
or basically everywhere they go. We have an extensive range of products for
this purpose. Plastic or metal carry cages, fashion pet bags, pet sling bags
and pet backpacks. These products are available in many different sizes,
designs and materials. Our pet strollers for small dogs have been a great
success in the pet market in recent years.
Grooming is also
an important part of a dog's life. We supply shampoos and conditioners for
different types of dog coats. We have a huge range of brushes and combs to
handle different grooming requirements.
Dog wear has been
a fashionable item worldwide in recent years. Many dog lovers like to dress
their pets for occasions like Halloween and Christmas. We provide
OEM services for dog wear or our own design of dog wear and dog shoes.
To maintain a
clean environment for dog owner's homes, we have dog toilets and also puppy training
pads to go with toilets. Both items are good for training puppies and also for
older dogs where mobility may be a problem.
Order Quantity:
One of our
advantages in supplying you our Dog Products is that we have been in the business
for almost four decades. We have a massive range of quality products. Our
advantage is that we can supply a very large variety of products to you in smaller
volumes than you may be required to purchase elsewhere.
We are flexible in
order volume, orders can be done in LCL or full containers. MOQ can be
discussed accordingly.
OEM Service:
Many of our customers have great ideas for Dog Products.
If you have an idea, we are happy to discuss details with you, including volumes,
costs and OEM etc. We can also help with developing products.
Dog Products
Dog Products,Dog Bedding,Dog Safety Products,Wear-Resistant Pet Bowl
PERCELL PET SYSTEM CO., LTD , https://www.percell-pet.com