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Rat GDF5 Elisa Test Kit CAT E1638Ra For Tissue Homogenates / Cell Lysates

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Rat GDF5 Elisa Test Kit CAT E1638Ra For Tissue Homogenates / Cell Lysates

Rat GDF5 Elisa Test Kit CAT E1638Ra For Tissue Homogenates / Cell Lysates
Rat GDF5 Elisa Test Kit CAT E1638Ra For Tissue Homogenates / Cell Lysates

Large Image :  Rat GDF5 Elisa Test Kit CAT E1638Ra For Tissue Homogenates / Cell Lysates

Product Details:

Place of Origin: Shanghai, China
Brand Name: BT Lab
Certification: CE, ISO9001:2005, MSDS
Model Number: Cat.No E1638Ra

Payment & Shipping Terms:

Minimum Order Quantity: Negotiation
Price: Negotiation
Packaging Details: Wrapped with ice pack and styrofoam package
Delivery Time: 1-3 business days, bulk order within one week
Supply Ability: Western Union, T/T
Detailed Product Description
Uniprot No.: M0RAY4 Customized: Acceptable
Storage: 2-8°C Brand: BT Lab
Cat.No: E1638Ra Shipping: DHL/FedEX
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immunoassay test kits

Strong Sensitivity Rat GDF5 ELISA Test Kit Customized Rat Growth Differentiation Factor 5 ELISA Kit

 

Cat.No E1638Ra

Standard Curve Range: 10ng/L - 2000ng/L

Sensitivity: 5.06ng/L

Size: 96 wells

 

Intended Use

This sandwich kit is for the accurate quantitative detection of Rat Growth Differentiation Factor 5 (also known as GDF5) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.

 

Precautions

  • Prior to use, the kit and sample should be warmed naturally to room temperature 30 minutes.
  • This instruction must be strictly followed in the experiment.
  • Once the desired number of strips has been removed, immediately reseal the bag to protect the remain from deterioration. Cover all reagents when not in use.
  • Make sure pipetting order and rate of addition from well-to-well when pipetting reagents.
  • Pipette tips and plate sealer in hand should be clean and disposable to avoid cross-contamination.
  • Avoid using the reagents from different batches together.
  • Substrate solution B is sensitive to light, don’t expose substrate solution B to light for a long time.
  • Stop solution contains acid. Please wear eye, hand and skin protection when using this material. Avoid contact of skin or mucous membranes with kit reagent.
  • The kit should not be used beyond the expiration date.

 

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.
  • Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.

*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 (2400ng/L) with 120μl of standard diluent to generate a 1200ng/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 (1200ng/L) 1:2 with standard diluent to produce 600ng/L, 300ng/L, 150ng/L and 75ng/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:

 

1200ng/L Standard No.5 120μl Original Standard + 120μl Standard Diluent
600ng/L Standard No.4 120μl Standard No.5 + 120μl Standard Diluent
300ng/L Standard No.3 120μl Standard No.4 + 120μl Standard Diluent
150ng/L Standard No.2 120μl Standard No.3 + 120μl Standard Diluent
75ng/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
2400ng/L 1200ng/L 600ng/L 300ng/L 150ng/L 75ng/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.

 

Assay Procedure

1. Prepare all reagents, standard solutions and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature.

2. Determine the number of strips required for the assay. Insert the strips in the frames for use. The unused strips should be stored at 2-8°C.

3. Add 50μl standard to standard well. Note: Don’t add antibody to standard well because the standard solution contains biotinylated antibody.

4. Add 40μl sample to sample wells and then add 10μl anti-GDF5 antibody to sample wells, then add 50μl streptavidin-HRP to sample wells and standard wells (Not blank control well). Mix well. Cover the plate with a sealer. Incubate 60 minutes at 37°C.

5. Remove the sealer and wash the plate 5 times with wash buffer. Soak wells with at least 0.35 ml wash buffer for 30 seconds to 1 minute for each wash. For automated washing, aspirate all wells and wash 5 times with wash buffer, overfilling wells with wash buffer. Blot the plate onto paper towels or other absorbent material.

6. Add 50μl substrate solution A to each well and then add 50μl substrate solution B to each well. Incubate plate covered with a new sealer for 10 minutes at 37°C in the dark.

7. Add 50μl Stop Solution to each well, the blue color will change into yellow immediately.

8. Determine the optical density (OD value) of each well immediately using a microplate reader set to 450 nm within 10 minuets after adding the stop solution.

 

Referances

"6-Hydroxydopamine induces distinct alterations in GDF5 and GDNF mRNA expression in the rat nigrostriatal system in vivo."
Gavin A.M., Walsh S., Wyatt S., O'Keeffe G.W., Sullivan A.M.
Neurosci. Lett. 561:176-181(2014)

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