Strong Specificity ELISA Assay Kit for Human Interleukin 1 Beta Detection 96 Wells

Strong specificity ELISA Kit for Human Interleukin 1 Beta 96 wells

Cat.No E0143Hu

Standard Curve Range: 20pg/L - 6000pg/L

Sensitivity: 10.07pg/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.

Reagent Provided

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

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.

Reagent Preparation

All reagents should be brought to room temperature before use.

Standard Reconstitute the 120μl of the standard (8000pg/L) with 120μl of standard diluent to generate a 4000pg/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 (4000pg/L) 1:2 with standard diluent to produce 2000pg/L, 1000pg/L, 500pg/L and 250pg/L solutions. Standard diluent serves as the zero standard(0 pg/L). Any remaining solution should be frozen at -20°C and used within one month. Dilution of standard solutions suggested are as follows:

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-IL-B 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.

References

Andrei C., Margiocco P., Poggi A., Lotti L.V., Torrisi M.R., Rubartelli A.
Proc. Natl. Acad. Sci. U.S.A. 101:9745-9750(2004)

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

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