Strong Specificity Mouse Vegf Elisa Kit 96 Wells / 48 Wells 2 - 8°c For Storage

Mouse Specificity and Precision VEGF ELISA Kit 96 Wells 48 Wells

Cat.No E0114Mo

Standard Curve Range: 5ng/L - 1800ng/L

Sensitivity: 2.48ng/L

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.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

CV(%) = SD/mean x 100

Intra-Assay: CV<8%

Inter-Assay: CV<10%

Intended Use

This sandwich kit is for the accurate quantitative detection of Mouse Vascular Endothelial Cell Growth Factor (also known as VEGF) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.

Material Required But Not Supplied

• 37°C±0.5°C incubator
• Absorbent paper
• Precision pipettes and disposable pipette tips
• Clean tubes
• Deionized or distilled water
• Microplate reader with 450 ± 10nm wavelength filter

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.

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-VEGF 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.

Shanghai Korain Biotech Co. was built in 2010. As an creator in reagents and tools for life science, Korainbio provide researchers with tools and scientific support including 30,000 antibodies, 1000+ proteins and 5000 ELISA kits. We aim to be a leading provider with world-class level for the researchers all over the world. Our products line covers a set of research areas including Immunology, Neuroscience, Cancer, Kinases, Phosphatases and Cell Biology.

Bioassay Technology Laboratory (BT Lab) as a leading brand of Korainbio focused on helping researchers to optimize life science work and serves as a provider of test and developing services, including elisa tests, WB test and antigen development. Founding in 2010, our strategic focus has been on the development of enabling technologies in the research, development, manufacture and marketing of innovative immuno products and services based on molecular technologies.

Korain’s reagents are supported by superior processional team and a quality management system that is certified for CE and ISO. Our product development program for use in a variety of applications including:

ELISA and immunohistochemistry

• Immunoprecipitation
• Immunohistochemistry
• Immunofluorescence microscopy
• Immunohistochemistry
• Quantitative Multiplexing

About BT Lab

As a leading brand of Korainbio, Bioassay Technology Laboratory (BT Lab) offers a variety of cost-effective ELISA kits and sets to measure cytokines, chemokines, and soluble biomarkers consistently and reliably. Sandwich kit and Competitive that provide the core reagents our selection of ELISA products meets the demand of ELISA beginners to experts alike at a very economical price.

Worldwide Distributors

ELISA Kit Categories

Sandwich Kit
Sandwich kits are fully validated and ready-to-use, containing 96- well strip plates pre-coated with capture antibody to detect sample antigen.

Competitive Kit
In a competitive ELISA assay, sample antigen and labeled antigen compete for capture antibody binding. The more target protein there is in the sample, the less labeled antigen will be captured and the weaker the signal.

Qualitative Kit
Qualitative results provide a simple positive or negative result for a sample. In quantitative ELISA, the optical density (OD) of the sample is compared to a standard curve, which is typically a serial dilution of a known-concentration solution of the target molecule. It is suitable for detecting small antigens.

Advantages of our Sandwich kit

Without Diluting Sample

• Save assay time
• Wide detection range
• Reducing non-specific interactions between the sample matrix proteins

Streptavidin-biotin system

• Significant non-specific binding can be prevented by strepvidin
• Reducing interference from residual biotin in sample
• A non-covalent interaction with high affinity

Washing Step

• Only 5 times for one step

Pre-Coated plate

What We Provide

• Product datasheet, product image, MSDS, publican references
• Order by emailor phone.
• Quality Guarantee with ISO and CE certificate
• Within 36 hours to your inquiry
• Support by professional team who are specializes in life science.
• Delivery to more than 100 countries
• Special packaging according your request

What We Aim to

• Optimize your R&D with our tools
• Build a friendship with every single customer
• Devoted ourselves to providing researchers with professional support

If you’re interested in working with us to expand the market, please feel free to contact us by: save@bt-laboratory.com