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What is a western blot test for HIV?
What is the principle of Western blotting?
How does the GS HIV-1 western blot kit work?
Confirmatory assays such as Western blot and immunoblot. Western blotting uses a blotting immunoassay technique to detect HIV-1 antibodies. In brief, HIV viral lysate is run on a polyacrylamide gel so that the viral proteins separate out according to their molecular weight; these proteins are then blotted on a nitrocellulose membrane.
- Principle
- Detection Can Be Done by Other Methods Such as
- Uses
Western blotting (protein blotting or immunoblotting) is a rapid and sensitive assay for the detection and characterization of proteins. It is based on the principle of immunochromatography where proteins are separated into polyacrylamide gel according to their molecular weight. The protein thus separated are then transferred or electrotransferred ...
Colorimetric detection
It depends on the incubation of the western blot with a substrate that reacts with the reporter enzyme (such as peroxidase) that is bound to the secondary antibody. This converts the soluble dye into an insoluble form of a different color that precipitates next to the enzyme and thereby stains the membrane. Development of the blot is then stopped by washing away the soluble dye. Protein levels are evaluated through spectrophotometry.
Radioactive detection
Radioactive labels do not require enzyme substrates, but rather, allow the placement of medical X-ray film directly against the western blot, which develops as it is exposed to the label and creates dark regions which correspond to the protein bands of interest. The importance of the radioactive detection method is declining due to its hazardous radiation because it is very expensive, health and safety risks are high, and ECL (enhanced chemiluminescence) provides a useful alternative.
Fluorescent detection
The fluorescently labeled probe is excited by light and the emission of the excitation is then detected by a photosensor such as a CCD camera equipped with appropriate emission filters which captures a digital image of the western blot and allows further data analysis such as molecular weight analysis and quantitative western blot analysis. Fluorescence is considered to be one of the best methods for quantification but is less sensitive than chemiluminescence. You can watch the following vide...
It is the most sensitive and specific test for determining the size and amount of protein present in any material.The confirmatory HIV test employs a western blot to detect anti-HIV antibodies in a human serum sample.A western blot is also used as the definitive test for Creutzfeldt-Jakob Disease, Lyme disease, hepatitis B infection, and HSV-2 (Herpes Type 2) infection.May 10, 2013 · The Western blot assay tests for antibodies that bind to fixed HIV proteins, which when exposed to a substrate create a pattern which can be read as positive, negative, or indeterminate. The IFA, a less frequently used alternative, mixes serum or plasma samples with T cells expressing HIV antigens to check for presence of antibodies.
- Julia Kang Cornett, Thomas J. Kirn
- 2013
May 28, 2023 · Applications of Western Blot. Western blotting is an excellent method with high sensitivity in order to detect a particular protein even in low quantity. Western blotting has been used in the clinical diagnosis of different diseases. The confirmatory test for HIV involves a western blot by detecting anti-HIV antibodies in the serum.
Western blot HIV test where first two strips are negative and positive controls followed by actual tests. The western blot is extensively used in biochemistry for the qualitative detection of single proteins and protein-modifications (such as post-translational modifications).
Method: HIV Antibody / HIV Western Blot Confirmatory Test. Serum/Plasma. Bio-Rad Laboratories HIV-1/HIV-2 plus O EIA (adopted July 2004) and Calypte HIV-1 Western Blot Kit. Important Information for Users. CDC periodically refines these laboratory methods.
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related to: western blot hiv principleCapture data from protein gels, nucleic acid gels, chemiluminescent western blots & more. Simplify western blot & gel documentation with the iBright Western Blot Imaging Systems.
Discover An HIV Treatment Option That May Help You Reach Undetectable. Learn More Now. Learn If You May Be Eligible To Save On Treatment. Visit The Official Patient Site Now.
Include a Lane for Positive Control in Your Western Blot. Find Your Control Today! A Better Way To Do Western Blot.
