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Shanghai Korain Biotech Co Ltd
7 Years
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| Brand Name : | BT Lab |
| Model Number : | Cat.No E0896Ra |
| Certification : | CE, ISO9001:2005, MSDS |
| Price : | Negotiation |
| Supply Ability : | Western Union, T/T |
| Delivery Time : | 1-3 business days, bulk order within one week |
96 wells Specificity and sensitivity Rat GHRL ELISA Kit
Cat.No E0896Ra
Standard Curve Range: 40ng/L - 12000ng/L
Sensitivity: 20.56ng/L
Size: 96 wells
Storage: Store the reagents at 2-8°C. For over 6-month storage refer to
the expiration date keep it at -20°C. Avoid repeated thaw cycles.
If individual reagents are opened it is recommended that the kit be
used within 1 month.
* This product is for research use only, not for use in diagnosis
procedures. It’s highly recommend to read this instruction entirely
before use.
Assay Principle
This kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate
has been pre-coated with Rat GHRL antibody. GHRL present in the
sample is added and binds to antibodies coated on the wells. And
then biotinylated Rat GHRL Antibody is added and binds to GHRL in
the sample. Then Streptavidin-HRP is added and binds to the
Biotinylated GHRL antibody. After incubation unbound
Streptavidin-HRP is washed away during a washing step. Substrate
solution is then added and color develops in proportion to the
amount of Rat GHRL. The reaction is terminated by addition of
acidic stop solution and absorbance is measured at 450 nm.
Specimen Collection
Serum Allow serum to clot for 10-20 minutes at room temperature.
Centrifuge at 2000-3000 RPM for 20 minutes.
Plasma Collect plasma using EDTA or heparin as an anticoagulant.
Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C
within 30 minutes of collection.
Urine Collect by sterile tube. Centrifuge at 2000-3000 RPM for
approximately 20 minutes. When collecting pleuroperitoneal fluid
and cerebrospinal fluid, please follow the procedures
above-mentioned.
Cell Culture Supernatant Collect by sterile tubes when examining secrete components.
Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect
the supernatants carefully. When examining the components within
the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the
cell concentration of approximately 1 million/ml. Damage cells
through repeated freeze-thaw cycles to let out the inside
components. Centrifuge at 2000-3000 RPM for approximately 20
minutes.
Tissue Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly
and weigh before homogenization. Mince tissues and homogenize them
in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or
freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20
minutes.
Note
Sample concentrations should be predicted before being used in the assay. If the sample concentration is not within the range of the standard curve, users must contact us to determine the optimal sample for their particular experiments.
Samples to be used within 5 days should be stored at 2-8°C. Samples should be aliquoted or must be stored at -20°C within 1 month or -80°C within 6 months. Avoid repeated freeze thaw cycles.
Samples should be brought to room temperature before starting the assay.
Centrifuge to collect sample before use.
Samples containing NaN3 can’t be tested as it inhibits the activity of Horse Radish Peroxidase (HRP).
Collect the supernatants carefully. When sediments occurred during storage, centrifugation should be performed again.
* Sample can't be diluted with this kit. Owing to the the material we use to prepare the kit, the sample
matrix interference may falsely depress the specificity and
accuracy of the assay.
Reagent Preparation
All reagents should be brought to room temperature before use.
Standard Reconstitute the 120μl of the standard (12800ng/L) with 120μl of
standard diluent to generate a 6400ng/L standard stock solution.
Allow the standard to sit for 15 mins with gentle agitation prior
to making dilutions. Prepare duplicate standard points by serially
diluting the standard stock solution (6400ng/L) 1:2 with standard
diluent to produce 3200ng/L, 1600ng/L, 800ng/L and 400ng/L
solutions. Standard diluent serves as the zero standard(0 ng/L).
Any remaining solution should be frozen at -20°C and used within
one month. Dilution of standard solutions suggested are as follows:
| 6400ng/L | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 3200ng/L | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 1600ng/L | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 800ng/L | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 400ng/L | Standard No.1 | 120μl Standard No.2 + 120μl Standard Diluent |
| Standard Concentration | Standard No.5 | Standard No.4 | Standard No.3 | Standard No.2 | Standard No.1 |
| 12800ng/L | 6400ng/L | 3200ng/L | 1600ng/L | 800ng/L | 400ng/L |
Wash Buffer Dilute 20ml of Wash Buffer Concentrate 30x into deionized or
distilled water to yield 500 ml of 1x Wash Buffer. If crystals have
formed in the concentrate, mix gently until the crystals have
completely dissolved.
Summary
1. Prepare all reagents, samples and standards.
2. Add sample and ELISA reagent into each well. Incubate for 1 hour at 37°C.
3. Wash the plate 5 times.
4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.
5. Add stop solution and color develops.
6. Read the OD value within 10 minutes.
References
Baldanzi G., Filigheddu N., Cutrupi S., Catapano F., Bonissoni S.,
Fubini A., Malan D., Baj G., Granata R., Broglio F., Papotti M.,
Surico N., Bussolino F., Isgaard J., Deghenghi R., Sinigaglia F.,
Prat M., Muccioli G., Ghigo E., Graziani A.
J. Cell Biol. 159:1029-1037(2002)
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