Western blot Secondary Antibody

Protein ladders are a set of standards known as molecular weight proteins that are utilized to identify the approximate size of a protein molecule run on a PAGE gel electrophoresis. The challenges in running the ladders are the choice of appropriate protein standard as it is used as visual evidence of protein migration, transfer efficiency, and positive control. Suitable protein markers can be selected on the basis of required properties and applications, i.e., fluorescent ladder, IEF, 2D SDS-PAGE ladder, natural ladder with an isoelectric point, and optimized ladders for Western Blot chemiluminescence detection. The key factors for running a distinct protein ladder are buffer conditions, charge/voltage at migration time, and the gel's concentration.

Protein ladders are a set of standards known as molecular weight proteins that are utilized to identify the approximate size of a protein molecule run on a PAGE gel electrophoresis. The challenges in running the ladders are the choice of appropriate protein standard as it is used as visual evidence of protein migration, transfer efficiency, and positive control. Suitable protein markers can be selected on the basis of required properties and applications, i.e., fluorescent ladder, IEF, 2D SDS-PAGE ladder, natural ladder with an isoelectric point, and optimized ladders for Western Blot chemiluminescence detection. The key factors for running a distinct protein ladder are buffer conditions, charge/voltage at migration time, and the gel's concentration.

Get tips on using CD29 (Integrin beta 1) Monoclonal Antibody (eBioHMb1-1 (HMb1-1)), APC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - CD29/β1-Integrin

Get tips on using MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), FITC, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - MHCII

Get tips on using MHC Class II (I-A/I-E) Monoclonal Antibody (M5/114.15.2), eFluor 450, eBioscience™ to perform Flow cytometry Anti-bodies Mouse - MHCII

Get tips on using WesternSure® Pre-stained Chemiluminescent Protein Ladder to perform Protein Ladder Immunofluorescence

Get tips on using DYKDDDDK Tag (D6W5B) Rabbit mAb (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #14793 to perform ChIP Anti-bodies FLAG

The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.

The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.

The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.