HMGB (High Mobility Group Box) ELISA Kits are designed to quantitatively measure the levels of HMGB proteins in various biological samples. HMGB proteins are non-histone nuclear proteins that play a critical role in DNA architecture, regulation of transcription, and cellular responses to damage. These kits are widely used in research related to inflammation, cancer, and other diseases where HMGB proteins are implicated.
Content
1. Principle of the Assay
HMGB ELISA Kits are based on the sandwich ELISA principle. Specific antibodies against HMGB proteins are used to capture and detect the target protein from the sample. The detection is typically performed using a colorimetric or chemiluminescent substrate that produces a measurable signal proportional to the amount of HMGB present in the sample.
2. Components
- Microplate: Pre-coated with a monoclonal antibody specific to HMGB.
- Standards: Recombinant HMGB protein standards for generating a standard curve.
- Detection Antibodies: Biotinylated or enzyme-conjugated secondary antibodies specific to HMGB.
- Substrate Reagents: TMB (3,3',5,5'-Tetramethylbenzidine) for colorimetric detection or luminescent substrates for chemiluminescent detection.
- Buffers: Washing buffer, sample diluent, assay diluent, and stop solution.
3. Assay Procedure
- Sample Preparation: Prepare biological samples such as serum, plasma, cell culture supernatants, or tissue extracts.
- Plate Setup: Add standards and samples to the wells of the microplate.
- Incubation: Incubate the plate to allow HMGB proteins to bind to the capture antibody.
- Washing: Remove unbound materials by washing the plate.
- Detection: Add the detection antibody and incubate, followed by another wash.
- Signal Development: Add the substrate solution and incubate until color or light is developed.
- Measurement: Measure the absorbance or luminescence using a plate reader.
- Data Analysis: Compare the sample readings to the standard curve to determine HMGB concentrations.
4. Applications
- Inflammation Research: Studying the role of HMGB in inflammatory responses and diseases such as sepsis.
- Cancer Research: Investigating HMGB expression and its involvement in tumor progression and metastasis.
- Cell Biology: Understanding the function of HMGB proteins in chromatin structure and gene regulation.
- Diagnostics: Developing potential diagnostic tools for diseases associated with altered HMGB levels.
5. Advantages
- High Sensitivity and Specificity: Accurate detection of HMGB proteins with minimal cross-reactivity.
- Quantitative Analysis: Provides precise quantification of HMGB levels in various sample types.
- Ease of Use: Simple and standardized protocols suitable for routine laboratory use.
- Versatility: Applicable to a wide range of research areas and sample types.
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