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  1. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...increasing the amount of SDS while decreasing the amount of alcohol. The increased SDS will increase the negative... subcellular compartment, you may want to try an SDS-containing detergent like RIPA. You can try a few...transfers, you can adjust the amounts of alcohol and SDS in your transfer buffers. To slow down smaller proteins...the amount of alcohol and decrease the amount of SDS, with a decreased transfer time. Alcohol will slow...proteins to stick to the membrane by stripping away the SDS. To speed up larger proteins and prevent them from...the current.  Pro tip! You need both alcohol and SDS for your proteins to migrate…so don’t try eliminating... transfer time Increase voltage Increase SDS/decrease alcohol in transfer buffer Small proteins...

  2. Hot Plasmids - February 2022

    Type
    Blog Post
    ...purification. Figure 3: The Penn State Ladder run on SDS-Page gels. A: individually expressed proteins run...are purified via histidine tags, are optimized for SDS-PAGE gels and can be detected on Western blots via...

  3. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...electrophoresis (SDS-PAGE) is the first step of a western. To prepare the samples for SDS-PAGE, measure ...(81)90281-5 Pillai-Kastoori L, Heaton S, Shiflett SD, Roberts AC, Solache A, Schutz-Geschwender AR (2019...

  4. Antibody Production

    Type
    Collection
    ...staining, the sample is separated via a denaturing SDS-PAGE gel alongside serial dilutions of a standard... Proteins are extracted from cells, separated by SDS-PAGE, transferred to a membrane, and labeled with...

  6. Technical Design of a Western Blot

    Type
    Blog Post
    ...in the cytoplasm or whole-cell lysate. A harsher, SDS-containing lysis buffer, like RIPA, should be considered...Bradford assays are not compatible with detergents like SDS, while BCA assays are not with denaturing agents ...proteins To denature your proteins, you'll want to use SDS and either DTT or β-mercaptoethanol (BME). DTT is... Table 1 lists the most common types of gels for SDS-PAGE running conditions and will cover most western...

  8. p53 Pathway

    Type
    Collection
    ...observed molecular weight (53 kDa) when run on an SDS-PAGE gel. Cancer Impact p53 is the most frequently...display early ageing-associated phenotypes. Tyner SD, Venkatachalam S, Choi J, Jones S, Ghebranious N,...

  14. Hot Plasmids Spring 2024

    Type
    Blog Post
    ... v1, vector 1; v2, vector 2; BD, binding domain; SDS, splice donor site; SAS, splice acceptor site; pA...

  17. Western Blot

    Type
    Protocol
    ...β-mercaptoethanol 4X protein loading buffer Precast SDS-PAGE gel SDS-PAGE running buffer Prestained protein ladder...the gel into the chamber of the SDS-PAGE gel tank. Pro-Tip Different SDS-PAGE gel tanks will have different...protein concentration in the lysate, running a precast SDS-PAGE gel, and immunoblotting. Protocols...protein concentration in the lysate, running a precast SDS-PAGE gel, and immunoblotting. Sharing speeds science...General Guidelines Acrylamide percentage varies in SDS-PAGE gels. Use lower percentage acrylamide when immunoblotting...2022 Workflow Timeline Day 1: Prepare lysates, run SDS-PAGE, transfer, block, incubate with primary antibody...protein concentration and prepare the lysate for SDS-PAGE Determine the protein concentration using a ...

  18. Antibody Guide

    Type
    Collection
    ...identify denatured proteins separated by size on an SDS-PAGE gel. Though relative quantification can be done...proteins, using heat and/or chemicals, and run on an SDS-PAGE gel to separate proteins by size. Transfer proteins...
Showing: 1 - 20 of 38 results