--- title: Colorimetric?  Fluorometric?  How to choose the right platform for your ELISA description: Choose a clear plate or a black plate for your next ELISA assay based on knowledge, not hunches! Read on for answers! --- **Please Note:** MBL International will be shutting down its operations effective December 31, 2024. Distribution of MBL products in the United States will be transferred to Cosmo Bio US on January 1st while European Distributors will remain unchanged. For any US inquiries regarding orders or support during this transition, reach out to Cosmo Bio: [https://www.cosmobiousa.com/](https://www.cosmobiousa.com/). For Non-US inquiries reach out to MBL in Japan: [https://www.mblbio.com/](https://www.mblbio.com/). 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[Home](https://blog.mblintl.com/) - [Blogs](https://blog.mblintl.com) - Colorimetric?  Fluorometric?  How to choose the right platform for your ELISA # Colorimetric?  Fluorometric?  How to choose the right platform for your ELISA ### Topics - Select Topic - 3D Cell Culture Advancements in Metabolism Research Allergy Antibodies Antibody Antibody Reagents Antibody-Peptide Conjugates Apoptosis Assay and Kits Autoimmune Disease Autophagy BRIC Benefits of Tetramer Assays Biomarker Cancer Cell Biology Cell Culture Reagents Cell Stabilization Custom Peptide Synthesis Cytokine Dextramer Staining Drug Discovery ELISA Epigenetics Epitope Tag Exosome Exosomes FCM Fc blocker Flow Cytometry Flow Cytometry Antibodies Fluoppi Fluorescent Proteins Future of Tetramer Technology Gastric Inhibitory Peptide Gene Regulation Gene Therapy Green Fluorescent Protein IL-18 Immune Response Immunology Inflammation Isotype Control Kits and Assays LC3 Liquid Biopsy MHC MHC Alleles MHC Class II Tetramers MHC Peptides Contribute MHC Tetramer MHC Tetramer Protocol MHC tetramer advancements Metabolism Mitophagy NCP Neuroscience Neuroscience Reagents Organoid PCSK9 PCSK9 ELISA Kit Peptide Peptide Exchange Peptide Synthesis Methods Peptide Synthesis Service Peptide-Infused Skin Care Products Preservation Purify exosome Quality Control QuickSwitch Quickswitch™ Custom Tetramer Kits RBP RIP Chip RNA Recombinant Protein Production Recombinant Proteins Regulation RiboTrap S100 Proteins SARS-CoV-2 T-Cell Detection TCR Tetramers Tetramer Tetramer Analysis Tetramer Assay Tetramer Assay Protocol Tetramer Staining TransFix Vectors Western Reagents atg8 covid-19 gfp keima red kinases mitochondria cell death p62 pathways phosphorylation pro IL-18 reagents rfp t cell tuberculosis ultracentrifugation vaccine ### Top Posts ### Subscribe to our Blogs Published by [Bindi M. Doshi, PhD](https://blog.mblintl.com/author/bindi-m-doshi-phd) on Sep 21, 2015 11:41:52 AM What happens when you use a black well plate for a fluorometric ELISA assay reading?  What happens when you use a clear well plate for a colorimetric ELISA assay reading?  Well the short answer is that you made the right plate decision! The concept of an ELISA assay is pretty straight forward, right? You have a sample (maybe some cells) and you want to know if something interesting is going on with your protein Y within that cell population.  You can either create your own ELISA (lots of work) or buy an ELISA kit (little work).  But what does it mean if a kit says it is colorimetric or fluorometric? For a colorimetric assay, energy is absorbed.  You are able to visually see which wells had a reaction and which wells did not.  Sometimes, it’s a nice blue color or a nice yellow color- you get the gist.  The concept of the assay is this: You have a plate that has a particular antibody bound to the wells.  You add your sample.  Maybe your sample binds to this antibody.  Then you add a detection antibody and a substrate to react with that detection probe.  After adding a stop solution, you can take readings to see how much of a particular analyte is in your sample.  For this assay, it is OK to use clear wells because the reaction occurs with no bleed through.  The darker the color, the greater the analyte in that well.  Pretty gratifying.  Only one wavelength is needed, however, some labs prefer to use two, one wavelength for the measurement reading and anther wavelength for reference.  This reference wavelength can compensate for well to well variations not having to do with the experiment (such as plastic difference, bubbles, etc.).  (However, if you have enough replicates, and you feel your physical well properties don’t vary too much, you may not need to run a reference wavelength.)  So in this case, you would subtract the reference values from the measurement value to give you the “real” value. A fluorometric assay is very similar, you add a substrate to your cells and then detect a fluorescence response using a plate reader.  You can think of it as energy is emitted.  In order to have the most accurate reading, the fluorescence of each individual cell needs to be measured and there should not be any cross over between the cells.  Hence, the reason to use opaque plates with this type of assay.  You want to reduce the amount of light scatter and a black plate will meet this need.  Two wavelengths are needed, one for excitation, one for emission. Fluorometric assays are a little more sensitive than colorimetric assays and they are able to detect more of an analyte than colorimetric assays.  They allow for a very high reading to be measured by widening the dynamic range.  Some labs will combine both techniques by using a clear bottom plate.  This allows for measuring a fluorescent probe for one analyte and a colorimetric probes for a second analyte. Regardless of your preference for colorimetric, fluorometric, or both, MBL has a product that can meet your needs.  Below are just a few examples: | Colorimetric | Fluorometric | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | | [4800](https://products.mblintl.com/products/4800/) | APOPCYTO™ Caspase-3 Colorimetric Assay Kit |   | [CY-1358](https://products.mblintl.com/products/cy-1358/) | CycLex® Protein Phosphatase LMW-PTP/ACP1 Fluorometric Assay Kit |   |   |   | | [4805](https://products.mblintl.com/products/4805) | APOPCYTO™ Caspase-8 Colorimetric Assay Kit |   | [CY-1355](https://products.mblintl.com/products/cy-1355/) | CycLex® Cdc25 Combo Protein Phosphatase Fluorometric Assay Kit |   |   |   | | [4810](https://products.mblintl.com/products/4810) | APOPCYTO™ Caspase-9 Colorimetric Assay Kit |   | [CY-1158V2](https://products.mblintl.com/products/cy-1158v2/) | CycLex® HDAC8 Deacetylase Fluorometric Assay Kit Ver.2 |   |   |   |   [![Contact Us!](https://no-cache.hubspot.com/cta/default/424171/6abbd33e-e1c2-4474-bc75-5f4529bb948a.png)](https://cta-redirect.hubspot.com/cta/redirect/424171/6abbd33e-e1c2-4474-bc75-5f4529bb948a) [![p53, Apoptosis, Autophagy and Other Pathway Posters](https://no-cache.hubspot.com/cta/default/424171/f8f49ae8-5e5a-4304-856d-883a362f1f80.png)](https://cta-redirect.hubspot.com/cta/redirect/424171/f8f49ae8-5e5a-4304-856d-883a362f1f80)   Tags: [Kits and Assays](https://blog.mblintl.com/topic/kits-and-assays) --- [Back to Blog](https://blog.mblintl.com) --- ### Comments Or Suggestions ## Related Articles ### Sign up for our latest news and promotions. 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