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5 × SDS Loading Buffer

    • 5 × SDS Loading Buffer-Elabscience
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    • 5 × SDS Loading Buffer-Elabscience
    • 5 × SDS Loading Buffer-Elabscience

      Catalog number:E-BC-R288

      Size:
      • 2 mL
      • 5 mL
      • 10 mL
      Qty:
      - +
      Price: $20

      Lead Time:Order now, ship in 3 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Introduction

      This product is a loading buffer for protein samples with SDS-PAGE electrophoresis. The SDS contained in the product can be combined with the protein to form a SDS-protein complex, which bring a large amount of negative charge to the protein; SDS can break intramolecular and intermolecular hydrogen bonds, and destroy the secondary and the tertiary structure of protein. The DTT contained in the preparation can break the disufide bond between the cysteine residues, destroy the structure between the proteins, and eliminate the difference between the protein structures. Ultimately, the rate of protein migration in the SDS-PAGE is only related to its molecular weight. Bromophenol blue is used as an indicator for electrophoresis to determine the progress of electrophoresis.

       

      Components

       

      Cat

      Products

      2 mL

      5 mL

      10 mL

      Storage

         E-BC-R288

         5 × SDS Loading Buffer

      1 mL × 2

      1 mL × 5

      1 mL × 10

      -20°C

          Manual

      1 copy





      Instructions

      1.Dissolve 5 × SDS Loading Buffer at room temperature or with water bath.

      2.For each 20 µL protein samples, add 5 µL 5 × SDS Loading Buffer and mix them well.

      3.Heat at 95~100℃ for 10 min to fully denatured the proteins.

      4.Centrifuge at 12000 rpm for 2 min, and collect the supernatant.

      5.Take 10~20 µL supernatant and add it into the SDS-PAGE gel well.

       

      5 XSDS Loading Buffer Components

      0.25 M Tris•HCl (pH6.8), 0.5 M DTT, 10% SDS, 0.5% BPB, 50% Glycerin.



      Storage

      Store at -20°C for 6 months.

       

      Cautions

      1.When the product is stored at -20°C, SDS precipitation may occur. Please dissolve it in warm water before use. Please store them properly according to its usage.

      2.If there is still a relatively viscous or viscous translucent object after sample boiling, please extend the boiling time or add 1 × SDS loading buffer and boil the sample again, so that the genomic DNA bound to the protein can be partially broken to reduce the generation of sample viscosity.

      3.This product contains DTT, which is slightly irritating and has certain toxicity.

      • Show all (1)
      • Reviews (0)
      • Q&A (1)
      Q

      M************* Submitted [ Sep 17 2022 ]

      Asked: Hello dear .. i hope this finds you well . Regarding this loading buffer , i use it recently and i faced some concerns . in directions number 2 , by protein sample did you mean sample alone or diluted with distilled water ? i used it according to BCA protein calculation ( 30ug/well) so i add ( 5.87ul of pt sample + 14.13ul dH2O ) to be 20ul + 5ul of loading buffer . Actually i get high bands intensity , i am doubt in this issue .. Kindly please guid me to overcome this . Best Regards .

      Reply

      A

      adminSubmitted [ Sep 17 2022 ]

      Answered: Hi Dear, Thanks for your feedback. Whether the protein sample needs to be diluted or not needs to be determined according to the concentration detected by BCA. If the extracted protein concentration is greater than 5mg/mL, it can be diluted appropriately with PBS to make the protein concentration around 4mg/mL before sample preparation. If the extracted protein concentration is lower than 5mg/mL, loading buffer can be directly applied to prepare samples according to the proportion specified in the instructions.

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