Tilt the tray slightly and add the specimen(s) where the buffer is collected at the lower end of each well as per the next step. Add 2 ml of blotting buffer to each well.Failure may result in watery marks on developed strips for some specimens.) Incubate the strips for 1 to 2 minutes at room temperature (25 ± 3☌) on a rocking platform (speed of 12 to 16 cycles per minute).Include strips for Strong Reactive, Weak Reactive, and Non-Reactive controls. Using forceps, carefully remove a required number of strips from the tube and place numbered sides up into each well.Add 2 ml of diluted wash buffer to each well.Overnight Assay ( long procedure) Rapid Assay Procedure Test procedure according to the manual of MP Diagnostics for HIV BLOT 2.2 It can be further completed by either chromogenic detection or.Protein transfer to a membrane: membrane stain and blocking and washing.Gel electrophoresis of protein: It may be performed by gel stain or.Sample preparation ( protein extraction).Paper towel, adhesive tape, worksheet (non-absorbent white paper).Sodium hypochlorite for decontamination.Aspirator with sodium hypochlorite trap. Precision Pipettes ranging from 20µl to 1000µl dispensing volume.Rocking platform (designed with a rocking speed range of 12 to 16Ĭycles per minute, and which moves through a 5° to 10° tilt to wash.Incorporated with HIV-1 viral lysate, a.The sodium dodecyl sulfate (SDS) Polyacrylamide gel electrophoresis (PAGE) technique is a prerequisite for Western blotting. A technique for detecting specific proteins separated by electrophoresis by the use of labeled antibodies. In Western blotting, the molecule of interest is a protein and the probe is typically an antibody raised against that particular protein. It is an Immunoblotting technique that relies on the specificity of binding between a molecule of interest and a probe to allow detection of the molecule of interest in a mixture of many other similar molecules. Western blot: It is applicable for transferring protein andĮastern blot: It is a biochemical technique used to analyze protein post-translational modifications (PTM) and is most often used to detect carbohydrate epitope. Northern blot: It uses for transferring RNA. Southern blot: It uses for transferring DNA. Blot is a technique for transferring DNA, RNA, and proteins onto a carrier so they can be separated, and often follows the use of gel electrophoresis. It is also called immunoblotting and it is a technique for detecting specific proteins separated by electrophoresis by the use of labeled antibodies. Western blot test is a confirmatory test of HIV-AIDS and it is positive as shown above image.
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