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WB37 HeLa Cell Pellet

HeLa Cell Pellet MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
WB37
  
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      Overview

      Replacement Information
      Description
      Overview

      This product has been discontinued.





      A human adenocarcinoma of the cervix. HeLa cells have been reported to contain human papilloma virus 18 (HPV-18), low expression of p53, and normal levels of Rb. Useful as a positive control for Western blotting of many different proteins, particularly cytokeratins, proteases, and those proteins involved in the apoptotic process.
      Catalogue NumberWB37
      Brand Family Calbiochem®
      References
      Product Information
      FormFlash-frozen cell pellet
      PreservativeNone
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Immunoprecipitation
      Application CommentsSuggested Preparation for Use

      Laemmli Sample Buffer (SDS-PAGE)
      Resuspend the frozen cell pellet by adding 1.0 ml of Laemmli sample buffer (containing fresh 2-mercaptoethanol, 2-ME, at a final concentration of 500 mM) to the vial containing the cell pellet followed by gentle vortexing. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Heat the sample for 5 min at 90°C. Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).

      Lane's Sample Buffer (IP/Immunoblot applications)
      Resuspend the frozen pellet by adding 1.0 ml of Lane's lysis buffer (150mM NaCl, 50 mM Tris-HCl, pH 8.0, 5 mM EDTA, and 1% NP40) to the vial containing the cell pellet followed by gentle vortexing. Fresh 2-mercaptoethanol, 2-ME, must be added to the vial at a concentration of 500 mM prior to heating the sample for 5 min at 90°C. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Dry Ice Only
      Toxicity Standard Handling
      Storage ≤ -70°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsStore at -70°C until lysed as described below. Following resuspension, store unused material in 25 μl aliquots at -70°C until needed. Do not expose to repeated cycles of freezing and thawing.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      WB37 0

      Documentation

      HeLa Cell Pellet Certificates of Analysis

      TitleLot Number
      WB37
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision25-October-2007 RFH
      DescriptionThe Calbiochem® Cell Pellets are a collection of frozen cell pellets representing cell lines most commonly used for internal controls in immunoblotting and IP/immunoblotting applications. Cells are grown under standard conditions to a density of approximately 80-85% confluency, washed extensively to remove excess culture medium derived proteins (predominantly serum albumin) and then pelleted and flash frozen. Pellets are designed to be lysed by the addition of Laemmli sample buffer for SDS-PAGE applications. Alternatively, the pellet may be lysed using alternative buffers such as Lane's Buffer or specific buffers with lower concentrations of detergent for IP/immunoblotting applications. Vial containing 10 X 106 of the indicated cell type, flash frozen at -70°C.
      BackgroundHeLa cells are derived from a human adenocarcinoma of the cervix and have been reported to contain human papilloma virus 18 (HPV-18), low expression of p53 and normal levels of Rb. HeLa is commonly used as a source of many different proteins, particularly cytokeratins, proteases, and those proteins involved in the apoptotic process. To preserve phosphorylation, orthovanadate should be included in the lysis buffer.
      FormFlash-frozen cell pellet
      PreservativeNone
      CommentsSuggested Preparation for Use

      Laemmli Sample Buffer (SDS-PAGE)
      Resuspend the frozen cell pellet by adding 1.0 ml of Laemmli sample buffer (containing fresh 2-mercaptoethanol, 2-ME, at a final concentration of 500 mM) to the vial containing the cell pellet followed by gentle vortexing. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Heat the sample for 5 min at 90°C. Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).

      Lane's Sample Buffer (IP/Immunoblot applications)
      Resuspend the frozen pellet by adding 1.0 ml of Lane's lysis buffer (150mM NaCl, 50 mM Tris-HCl, pH 8.0, 5 mM EDTA, and 1% NP40) to the vial containing the cell pellet followed by gentle vortexing. Fresh 2-mercaptoethanol, 2-ME, must be added to the vial at a concentration of 500 mM prior to heating the sample for 5 min at 90°C. (Alternatively dithiothreitol (DTT) to a final concentration of 100 mM may be substituted for the 2-ME). Apply 15-25 µl to each lane of the gel. This is a starting amount, as actual amounts for optimal protein detection need to be experimentally determined for every antibody. Freeze the unused material in 25 µl aliquots and store at -70°C. (Before using the frozen samples, fresh 2-ME or DTT should be added and the sample heated for 5 min at 90°C).
      Storage Avoid freeze/thaw
      ≤ -70°C
      Do Not Freeze Ok to freeze
      Special InstructionsStore at -70°C until lysed as described below. Following resuspension, store unused material in 25 μl aliquots at -70°C until needed. Do not expose to repeated cycles of freezing and thawing.
      Toxicity Standard Handling