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#dotblot #RohitMane #ScientistRacademy Dot Blot is a cheaper, easier and faster technique to detect the presence of Proteins and Nucleic Acids in a biological sample. Due to the simplicity of the technique it widely used as a ideal diagnostic tool. This post covers the Protein Dot Blot Technique. Dot Blot Principle Dot Blot works based on the immunodetection principle for identifying specific protein. Dot Blot - PrincipleDot Blot - Technique: Overview Proteins are immobilized on to a protein binding membrane, PVDF or Nitrocellulose. After Protein Immobilization membrane is blocked to avoid non-specific binding, using blocking buffer. Immobilized protein is then incubated with a primary antibody (for 30mins to 1 hour). The antibody is specific to the protein and is visualized either with a specific tag or with a secondary antibody.  The secondary antibody generally will carry a tag that allow the visualization of protein. Blots can be visualized using methods such as Chemiluminescence or Fluorescence. Common Tags used in Dot Blots are enzymes conjugated to the secondary antibody. Upon addition of substrate, enzyme acts on the substrate to develop colour. Commonly preferred enzymes are HRP(Horse Radish Peroxidase) and Alkaline Phosphatase (AP) Advantages of Dot Blot over Western Blot Non-Electrophoresis Technique: Dot Blot is non-electrophoresis technique, means that one need not do protein electrophoresis. Proteins can directly immobilized on to the membrane. Simple & Easy Method: No messy work of PAGE run and gel to membrane transfers, proteins are directly immobilized on membrane, making this method much easier and faster. Time: It is a faster method, takes ~3hours, whereas western blot may even take up to 2 days. Including the protein electrophoresis and gel to membrane transfer optimization. Dot Blot is not as informative as Western Blot, but it is inexpesive, quick and easy method for the simple protein detection.  dot blot technique by shomus biology dot blot technique in hindi

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Vince Neil Vince Neil, the legendary front man of Mötley Crüe, is an international superstar, accomplished businessman and heartfelt philanthropist. His unmistakable voice leads some of the greatest rock songs of a generation including “Wild Side”, “Shout at the Devil,” “Home Sweet Home,” “Girls, Girls, Girls,” “Dr. Feelgood,” “Kickstart My Heart,” “Same Ol’ Situation,” and many others. Raised on the tough streets of Compton, California, Vince Neil fell in love with music as a child, developing a broad vocabulary that ranged from classic Motown to Hard Rock. As a teenager, he began performing with local bands and soon discovered both a passion for the stage and a natural talent as a singer and performer. He fronted the popular band Rock Candy before reuniting with high school buddy Tommy Lee in 1981, establishing his spot in rock and roll history with the formation of Mötley Crüe. Presently, Neil remains primarily occupied answering the call to his immense following with shows internationally in conjunction with the culmination of his fourth solo album. Neil continues to perform all the classic Mötley Crüe hits with select cuts from his three solo albums. Additionally, this badass performer is working on new music and has hinted about performing some of it during this tour. Stephen Pearcy As the founder, lead singer and songwriter of the metal rock band RATT, Stephen Pearcy is credited with having been a pioneer of the early 80's Sunset Strip rock scene. Having led his creation to Mega Multi-Platinum Success Year after Year from 1984 to 1991. RATT climbed up the charts with singles from “Out of the Cellar”, soaring past Prince’s “Let’s Go Crazy”, Van Halen’s “Panama”, Madonna’s “Borderline” and David Bowies “Blue Jean”. RATT & Mickey RATT songs are featured regularly in film & TV; Point Break, Weird Science, The Golden Child, Academy Award winning movie The Wrestler, Emmy Award winning Stranger Things, Wicked City, Cobra Kai, The Goldberg’s, Young Sheldon... The Pearcy legacy is here to stay. His fans are loyal and always wanting more of him…after all, he is the original “Wanted Man”.

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