Choosing The Right ELISA Kit Can Help You Achieve More With Less Effort
1. What is ELISA Kit？
Enzyme-linked immunosorbent assay ( ELISA ) is the most widely used technology in enzyme immunoassay technology . The invention of ELISA Kit has helped scientific researchers save the tedious experimental process and improved scientific research efficiency. Choosing the right kit and combining good experimental skills will make your experimental efficiency twice as good with half the effort.
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2. How to choose an ELISA Kit?
2.1 Find the protein concentration range of the sample to be testedReference database: Search for reference values of target protein expression through databases such as NCBI, UniProt, and Pax-Db to ensure that the detection range of the kit is consistent with the reference value.
2.2 Application species of the kit and types of test samplesSpecies specificity: Samples from different species are usually not compatible unless otherwise specified in the kit.Sample type: Common sample types include serum, plasma (with different anticoagulants), cell supernatant and cell lysate . Different samples may require different kits. Make sure the kit is suitable for your sample type.
2.3 Detection range of the kitStandard curve range: Ensure that the concentration of the sample to be tested falls within the standard curve range of the kit. For high-concentration samples, it is recommended to conduct a preliminary experiment to determine the appropriate dilution factor. Due to individual differences between samples, it is generally recommended to select more than 4 samples for preliminary experiments to determine the optimal dilution factor. 2.4 Sensitivity of the kitSelect a sensitivity suitable for the content of the indicator to be tested. For low-content indicators, a high-sensitivity kit is more suitable.
2.2 Application species of the kit and types of test samplesSpecies specificity: Samples from different species are usually not compatible unless otherwise specified in the kit.Sample type: Common sample types include serum, plasma (with different anticoagulants), cell supernatant and cell lysate . Different samples may require different kits. Make sure the kit is suitable for your sample type.
2.3 Detection range of the kitStandard curve range: Ensure that the concentration of the sample to be tested falls within the standard curve range of the kit. For high-concentration samples, it is recommended to conduct a preliminary experiment to determine the appropriate dilution factor. Due to individual differences between samples, it is generally recommended to select more than 4 samples for preliminary experiments to determine the optimal dilution factor. 2.4 Sensitivity of the kitSelect a sensitivity suitable for the content of the indicator to be tested. For low-content indicators, a high-sensitivity kit is more suitable.
3. ELISA usage tips
3.1 Sample preparationFresh samples: Use fresh samples as much as possible and avoid using samples that are hemolyzed, lipolyzed , stored for too long, or incompletely coagulated.Storage conditions: If the test cannot be performed immediately, the sample should be stored at 4°C within 5 days. Samples tested after 1 week should be frozen at low temperature to avoid repeated freezing and thawing.
3.2 Read the instructions carefullyFamiliarize yourself with the process: It is recommended to draw a simple step-by-step diagram, prepare reagents in advance, and prepare working solutions of concentrated reagents in advance .Batch consistency: Do not mix reagents from different batches and check the expiration date of each component.
3.3 Pre-coated ELISA plate Balance of reagent componentsRoom temperature equilibrium: The pre-coated ELISA strips and reagent components need to be equilibrated at room temperature for more than 30 minutes to ensure that the temperature in the reaction microwells reaches the required temperature to avoid missing weak positive samples.
3.4 Adding samples to be testedBatch operation: When there are too many samples, it is recommended to operate in batches and dilute them in advance.Sample addition tips: control the sample addition time to avoid errors; add vertically to the bottom of the well to avoid contact with the well wall; use disposable pipette tips to prevent cross contamination.
3.5 Incubation and washingIncubation conditions: Select the incubation time and temperature according to the instructions. To avoid uneven temperature, try to use a constant temperature incubator for incubation .Plate washing tips: When washing plates manually, tap the plates vertically to avoid cross contamination; when washing plates with a plate washer, check whether the rinse head is unobstructed. 3.6 Color RenderingColor development conditions: TMB is used as the substrate reaction solution. The required amount needs to be measured and added. It is forbidden to pour out all of it and then pour the rest back . React at 37°C for 20 minutes ( the incubation time can be shortened or extended as appropriate, but not more than 30 minutes) . The color development reaction needs to be protected from light.
3.7 Reading and data analysisRead the results in time: Read the results within 15 minutes after terminating the reaction. The microplate reader needs to be preheated for 15-30 minutes. Pay attention to choosing the appropriate wavelength for reading . 4. Data Analysis: Use Curve Expert software to create a standard curve.Download address : http://www.elkbiotech.cn/list/83.htmlFor software usage, please refer to: http://www.elkbiotech.cn/view/683.html
3.2 Read the instructions carefullyFamiliarize yourself with the process: It is recommended to draw a simple step-by-step diagram, prepare reagents in advance, and prepare working solutions of concentrated reagents in advance .Batch consistency: Do not mix reagents from different batches and check the expiration date of each component.
3.3 Pre-coated ELISA plate Balance of reagent componentsRoom temperature equilibrium: The pre-coated ELISA strips and reagent components need to be equilibrated at room temperature for more than 30 minutes to ensure that the temperature in the reaction microwells reaches the required temperature to avoid missing weak positive samples.
3.4 Adding samples to be testedBatch operation: When there are too many samples, it is recommended to operate in batches and dilute them in advance.Sample addition tips: control the sample addition time to avoid errors; add vertically to the bottom of the well to avoid contact with the well wall; use disposable pipette tips to prevent cross contamination.
3.5 Incubation and washingIncubation conditions: Select the incubation time and temperature according to the instructions. To avoid uneven temperature, try to use a constant temperature incubator for incubation .Plate washing tips: When washing plates manually, tap the plates vertically to avoid cross contamination; when washing plates with a plate washer, check whether the rinse head is unobstructed. 3.6 Color RenderingColor development conditions: TMB is used as the substrate reaction solution. The required amount needs to be measured and added. It is forbidden to pour out all of it and then pour the rest back . React at 37°C for 20 minutes ( the incubation time can be shortened or extended as appropriate, but not more than 30 minutes) . The color development reaction needs to be protected from light.
3.7 Reading and data analysisRead the results in time: Read the results within 15 minutes after terminating the reaction. The microplate reader needs to be preheated for 15-30 minutes. Pay attention to choosing the appropriate wavelength for reading . 4. Data Analysis: Use Curve Expert software to create a standard curve.Download address : http://www.elkbiotech.cn/list/83.htmlFor software usage, please refer to: http://www.elkbiotech.cn/view/683.html
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By following the above steps, you can more effectively select and use ELISA Kits, thereby improving the accuracy and efficiency of your experiments.