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Microplate Absorbance Reader

Instrument Images
BM0007

INSTRUMENT INCHARGE

Prof. Subhankar Paul

Professor

Protein conformation and assisted folding, Nanotechnology in cancer therapy, Biomolecule-based nanostructure development, Bone Tissue Engineering

BASIC INFORMATION

  • Make & Model : BioRad and Model No: I -Mark
  • Department : Biotechnology and Medical Engineering
  • Location : Structural Biology Laboratory
  • Category : Category - III
  • Status : Non-Functional

PRINCIPLE:

The iMark Microplate Reader operates on the basic principle of absorbance spectrophotometry, where the amount of light absorbed by a sample is measured to determine the concentration of a substance in that sample. Light Source:The microplate reader has a light source, typically a halogen lamp, which produces light across a range of wavelengths (usually in the UV-visible spectrum). Wavelength Selection:The light from the source is directed through an optical filter system that selects specific wavelengths of light depending on the needs of the assay. The user can select one or more wavelengths (e.g., 450 nm, 570 nm) depending on the absorbance peak of the analyte being measured. Light Interaction with the Sample:The selected light passes through each well of the 96-well microplate, where it interacts with the sample. If the sample contains a substance that absorbs light at the selected wavelength, the amount of light that passes through the sample will be reduced based on the concentration of the absorbing substance. Absorbance Measurement:After the light passes through the sample, a photodetector (typically a photodiode or photomultiplier tube) detects the amount of light that has passed through the sample. The detector compares the intensity of the incident light (before it interacts with the sample) and the transmitted light (after it passes through the sample). The difference in light intensity is used to calculate the absorbance of the sample. Absorbance Calculation:Absorbance is calculated using Beer-Lamberts law: ??=log0??) A=log( II 0) Where:A is the absorbance (no units), I0 is the intensity of light before passing through the sample (incident light), I is the intensity of light after passing through the sample (transmitted light). Data Output:The iMark Microplate Reader provides real-time absorbance data that is displayed on the connected computer or device. This data can then be used for further analysis, such as determining the concentration of an analyte using standard curves or calculating enzyme activity or cell viability.

APPLICATION:

ELISA (Enzyme-Linked Immunosorbent Assay):The iMark Microplate Reader is commonly used in ELISA assays for the detection and quantification of antibodies, antigens, and other proteins in biological samples. It allows researchers to measure absorbance at different wavelengths (e.g., 450 nm for many ELISA kits) to determine the concentration of the target analyte. Protein Quantification :The iMark Microplate Reader is used in assays such as the Bradford Assay, BCA Assay, and Lowry Assay for determining the concentration of proteins in samples. The reader measures the absorbance at specific wavelengths corresponding to the color changes in these assays. Cell Viability and Cytotoxicity Assays : It is widely used in cell-based assays to measure cell viability, proliferation, and cytotoxicity. For example, assays like the MTT assay, XTT assay, and Alamar Blue are commonly performed to assess the health and growth of cultured cells. The iMark Microplate Reader quantifies cell viability by measuring absorbance at specific wavelengths (e.g., 490 nm or 570 nm). Nucleic Acid Quantification:The microplate reader can be used for quantification of nucleic acids such as DNA and RNA. Specific assays like NanoDrop or A260/A280 ratio can be performed to assess the concentration and purity of nucleic acid samples. Enzyme Activity Measurement :It is used to measure the activity of enzymes through the change in absorbance of the reaction mixture. This is particularly useful in kinetic assays where the change in absorbance over time can be monitored at specific wavelengths to assess enzyme activity. Binding and Inhibition Studies:The iMark Microplate Reader is used in binding assays to study the interactions between ligands and receptors or to measure inhibition in enzyme or protein function. It can detect small changes in absorbance that occur as a result of these interactions. Immunoassays :Immunoassays such as competitive binding assays and sandwich immunoassays can be performed with the iMark Microplate Reader to detect various substances like hormones, drugs, infectious agents, or vitamins in clinical and research settings. Kinetic and Endpoint Assays: The iMark allows for both kinetic measurements (monitoring absorbance over time) and endpoint measurements (taking absorbance readings at a specific time point). This is useful in a wide range of biochemical and immunological assays where changes in absorbance indicate reactions or interactions. Metabolite and Hormone Analysis :In clinical research, the iMark Microplate Reader is used to measure various metabolites or hormones in biological fluids, such as serum or plasma, by using specific assay kits designed for these purposes. Biological Sample Analysis:The iMark Microplate Reader can be used to analyze various biological samples, such as serum, plasma, urine, and saliva, for a wide range of diagnostic tests and research applications.

EXPERIMENTS & CHARGES:

EXPERIMENT - 1

  • Name of Experiment : Microplate reader analysis
  • Condition of Analysis : As per MSDS and Cover letter
  • Service Units : 8 Sample
  • Academic & Educational Institution Charges : ₹ 118.00
  • R & D National Laboratory Charges : ₹ 177.00
  • Industry Charges : ₹ 354.00
  • International Charges : ₹ 1,770.00
  • Available for Sample Booking : No
  • Remarks :