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ELISA protocols and Troubleshooting guide for high quality and reliable data.Browse Boster Featured Products
ELISA (enzyme-linked immunosorbent assay) is a plate-based assay technique designed for detecting and quantifying peptides, proteins, antibodies, and hormones. In ELISA, an antigen must be immobilized to a solid surface and then complexed with an antibody that is linked to an enzyme. Detection is accomplished by assessing the conjugated enzyme activity via incubation with a substrate to produce a measurable product. The most crucial element of the detection strategy is a highly specific antibody-antigen interaction.
ELISAs are typically performed in 96-well (or 384-well) polystyrene plates, which will passively bind antibodies and proteins. The binding and immobilization of reagents makes ELISAs simple to design and perform. Having the reactants of the ELISA immobilized to the microplate surface enables easy separation of bound from non-bound material during the assay. This ability to wash away non-specifically bound materials makes the ELISA a powerful tool for measuring specific analytes within a crude preparation.
Enzyme-linked immunosorbent assays (ELISA) principles are very similar to other immunoassay technologies. ELISAs rely on specific antibodies to bind the target antigen, and a detection system to indicate the presence and quantity of antigen binding. In order to maximize the sensitivity and precision of the assay, the plate must be carefully coated with high-affinity antibodies – a process that Boster Bio has mastered.
Did you know that Boster, the leading ELISA manufacturers are providing research-grade CRO services? Outsource your ELISA experiments today and save your time and money.
Our 2300+ ELISA kits are validated in multiple sample matrices from serum and saliva to urine and feces, ensuring wide application ranges for you to select from. Boster's mission is to support research in areas such as immunology, neuroscience, cancer, and more by providing the high-quality ELISA kits needed to get better results. Here are the list of our most popular ELISA kit.
Comparisons on Direct, Indirect, Sandwich, Competitive ELISA
ELISAs can be performed with a number of modifications to the basic procedure: direct, indirect, sandwich or competitive. The key step, immobilization of the antigen of interest, can be accomplished by direct adsorption to the assay plate or indirectly via a capture antibody that has been attached to the plate. The antigen is then detected either directly (enzyme-labeled primary antibody) or indirectly (enzyme-labeled secondary antibody). The detection antibodies are usually labeled with alkaline phosphatase (AP) or horseradish peroxidase (HRP). A large selection of substrates is available for performing the ELISA with an HRP or AP conjugate. The choice of substrate depends upon the required assay sensitivity and the instrumentation available for signal-detection (spectrophotometer, fluorometer or luminometer).
Among the standard assay formats discussed and illustrated below, where differences in both cpture and detection were the concern, it is important to differentiate between the particular strategies that exist specifically for the detection step. However an antigen is captured to the plate (by direct adsorption to the surface or through a pre-coated "capture" antibody, as in a sandwich ELISA), it is the detection step (as either direct or indirect detection) that largely determines the sensitivity of an ELISA.
For direct detection, an antigen coated to a multi-well plate is detected by an antibody that has been directly conjugated to an enzyme. This detection method is a good option if there is no commercially available ELISA kits for your target protein.Hover to See More
For indirect detection, the antigen coated to a multi-well plate is detected in two stages or layers. First an unlabeled primary antibody, which is specific for the antigen, is applied. Next, an enzyme-labeled secondary antibody is bound to the first antibodyHover to See More
Sandwich ELISAs typically require the use of matched antibody pairs, where each antibody is specific for a different, non-overlapping part (epitope) of the antigen molecule. A first antibody (known as capture antibody) is coated to the wells.Hover to See More
Sandwich ELISAs typically require the use of matched antibody pairs, where each antibody is specific for a different, non-overlapping part (epitope) of the antigen molecule. A first antibody (known as capture antibody) is coated to the wells. The sample solution is then added to the well. A second antibody (known as detection antibody) follows this step in order to measure the concentration of the sample. This type of ELISA has the following advantages:
This ELISA kit is of competitive format. Competitive ELISA, also known as inhibition ELISA, is a surface/plate based assay, where the plate is coated with capture antibodies reactive to the molecule of interest.Hover to See More
This ELISA kit is of competitive format. Competitive ELISA, also known as inhibition ELISA, is a surface/plate based assay, where the plate is coated with capture antibodies reactive to the molecule of interest. The sample (containing native molecule of interest) and enzyme conjugated recombinant protein (the competing molecule) are added to the coated wells. Since the amount of enzyme conjugated molecule in each well is constant, the level of native molecule in the sample will determine the binding ratio of enzyme conjugated molecule vs. native molecule. After an incubation period, any unbound antibody is washed off. Enzyme substrate (for example, TMB for HRP) is added to each well and will be transformed into a blue precipitate, the amount of which is linearly proportional to the amount of enzyme in the well. The precipitate is then turned into yellow by adding the acid stop solution and the concentration of yellow precipitate is read at 450nm for light absorbance (O.D. value). The O.D. is then used to calculate the amount of molecule of interest in each well, by comparing each sample well against the standard curve. The standard curve is generated using the same principle but instead of adding samples, a series of recombinant molecules with known concentrations are added to 6-8 wells.
General ELISA Workflow, a step-by-step ELISA protocol
96-Well Plate Pre-Coated with Capture Antibody (Choose from Boster's ELISA Kit)
|Capture Ab Coating|
|Sample (Antigen) Incubation|
|Primary Ab Incubation|
|Secondary Ab Incubation|
ELISA data is typically graphed with optical density (or fluorescence) vs concentration to produce a sigmoidal curve as shown above.
This guide will teach you everything you need to become an ELISA expert, including a critical review of principles, all-in-one FAQs, and more.
Picokine ELISA kits are Boster Bio manufactured ELISA kits that have Picokine level sensitivity. Our ELISA kits come with over 20 years of manufacturing expertise and proprietary methods which provide the precision you need.
Boster Bio is proud to offer over 1,000 ELISA kits for a wide range of targets. With over 20 years of experience in antibody and ELISA kit manufacturing, Boster Picokine™ enzyme linked immunosorbent assay (ELISA) kits are guaranteed to be sensitive, specific, and stable. We rigorously validate every lot against a wide range of samples to ensure consistent, reliable results. Join over 14,000 scientists who put their trust in Boster Picokine™ ELISA kits.
Boster's Picokine™ ELISA kits are made with high affinity antibodies that can detect native form proteins with picogram and subpicogram level sensitivity.
Boster's QC department validates our ELISA kits against proteins in relevant superfamilies and proteins with similar immunogenicities to ensure specificity to the analytes of interests.
Our 2300+ ELISA kits are validated in multiple sample matrices from serum and saliva to urine and feces, ensuring wide application ranges for you to select from.
Boster has been serving the research community since 1993 and cited by 23,000+ publications. Our team of experts are dedicated to provide you the best customer service.
Cited by more than 1000+ publications
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Here are the 212 most popular ELISA kits.
|TNF Alpha elisa||IL6 ELISA||Cortisol ELISA||VEGF elisa|
|BDNF elisa||IFN Gamma elisa||Adiponectin elisa||IL-1 Beta elisa|
|IL10 ELISA||IL-8 elisa||Leptin elisa||IL2 ELISA|
|IL-12 elisa||Granzyme B elisa||MPO elisa||ADA elisa|
|APP elisa||TGF Beta 1 elisa||MAG elisa||IL4 ELISA|
|MMP-9 elisa||PLAT elisa||Cystatin C elisa||CCL2 ELISA|
|IL-17 elisa||PD-L1 elisa||APOE elisa||NGF elisa|
|CXCL10 elisa||PAI-1 elisa||S100B elisa||Galectin-3 elisa|
|EGF elisa||Fibronectin elisa||GM-CSF elisa||MMP-3 elisa|
|Cortisol elisa Kit||Insulin elisa Kit||IL33 ELISA||GDF-15 elisa|
|Resistin elisa||FGF21 elisa||AFP elisa||Angiopoietin-2 elisa|
|Clusterin elisa||P53 elisa||IDS elisa||Ferritin elisa Kit|
|MMP-1 elisa||OPN elisa||Endothelin 1 elisa||PCSK9 elisa|
|HGF elisa||G-CSF elisa||VWF ELISA||CXCL1 elisa|
|PD-1 elisa||Caspase 3 elisa||TIMP1 ELISA||P-Selectin elisa|
|Tissue Factor elisa||TRAIL elisa||Fetuin A elisa||Chemerin elisa|
|IL-15 elisa||COMP elisa||IL-22 elisa||ANG ELISA|
|CEA elisa||Periostin elisa||Galectin-9 elisa||MMP2 ELISA|
|TEK ELISA||Cathepsin B elisa||CXCL5 elisa||CXCL9 elisa|
|VEGFC ELISA||CCL17 elisa||CXCL13 elisa||IL-27 elisa|
|PEDF elisa||ADAMTS13 elisa||APOA1 elisa||Eotaxin elisa|
|M-CSF elisa||PLGF elisa||RANK elisa||Thrombomodulin elisa|
|MIA elisa||HE4 elisa||IL7 ELISA||PDGF-AB elisa|
|C-MET elisa||IL-1RA elisa||Renin elisa||FABP2 elisa|
|BMP2 ELISA||IL17C ELISA||LCN2 ELISA||MMP12 ELISA|
|Fractalkine elisa||CCL19 elisa||CCL21 elisa||Angiopoietin-1 elisa|
|Growth Hormone elisa||CCL18 elisa||THBS1 elisa||TSLP elisa|
|SHBG elisa||Hemopexin elisa||TIM-3 elisa||MMP7 ELISA|
|FAS elisa||TREM2 elisa||Myoglobin elisa Kit||FGF2 elisa|
|GDNF elisa||PTX3 elisa||TGF-Beta 2 elisa||Mesothelin elisa|
|Transthyretin elisa||DKK1 ELISA||FASL elisa||CCL4 elisa|
|MMP-8 elisa||OPG elisa||Rantes elisa||ACE elisa|
|CD40 elisa||CXCL11 elisa||Prostate Specific Antigen elisa Kit||FGF7 elisa|
|Midkine elisa||Uromodulin elisa||PROC elisa||IL11 ELISA|
|IL31 ELISA||IL-3 elisa||CCL8 elisa||MIF elisa|
|RBP4 elisa||TLR2 elisa||FABP4 elisa||B2M elisa|
|IL1A ELISA||LTA ELISA||ERBB2 ELISA||FAP ELISA|
|SERPINA1 ELISA||COL1A1 ELISA||IGFBP2 elisa||CCL22 elisa|
|CD163 elisa||MICA elisa||Progranulin elisa||FGF19 elisa|
|FOLR1 elisa||Syndecan-1 elisa||CEACAM1 elisa||MUC1 ELISA|
|Amphiregulin elisa||IL-5 elisa||Decorin elisa||S100A12 elisa|
|Tenascin-C elisa||IL-32 elisa||TFPI ELISA||IL18BP ELISA|
|AXL elisa||Aggrecan elisa||ALCAM elisa||ICAM1 ELISA|
|CCL20 ELISA||TIMP2 ELISA||TIMP4 ELISA||PROK1 ELISA|
|DPP4 ELISA||ANXA1 ELISA|
|Haptoglobin elisa||FGF23 elisa||CRP elisa||EPO elisa|
|SAA elisa||Elastase elisa||IL-13 elisa||S100A8 elisa|
|LBP elisa||PF4 ELISA||Stat3 ELISA||IL-23 elisa|
|CD14 elisa||CTLA4 elisa||LIF elisa||IL-21 elisa|
|CD30 elisa||S100A9 elisa||TREM-1 elisa||TrkB elisa|
|SAP elisa||Thrombopoietin elisa||SCF elisa||MFGE8 elisa|
|GAS6 elisa||CXCL2 ELISA||Sclerostin elisa||Mpl ELISA|
ELISAs can accurately assess soluble proteins in their native state, so they are ideal for samples such as urine or saliva. Check out the ELISA sample preparation guides to learn how to get the best results from your sample type.Learn our ELISA Sample Preparation
Learn stepwise ELISA protocols from reagent preparation to data analysis. Check out their differences to learn how they compare and their advantages and disadvantage.Learn our ELISA protocol
Get to know some ELISA troubleshooting tips with this Guide. It has some commonly encountered problems and solutions to ELISA.Check our ELISA troubleshooting tips
Learn how to perform ELISA data analysis. Get to know the different aspects to consider for more consistent and accurate ELISA data. Furthermore, we provide a step-by-step guide to create a standard curve for analysis.Browse ELISA Data analysis