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  1. Link

    products/cell-lysates/a549-whole-cell-lysate-ab7910.pdf

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Kits/ Lysates/ Other Lysates Whole Cell Lysates Human Other
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A549 whole cell lysate (ab7910)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (1)

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Overview

  • Product name

    A549 whole cell lysate
  • General notes

    Cell line: A549 (Human lung adenocarcinoma epithelial).
    Growth media: DMEM and 10% NCS (Newborn calf serum).

     

    A549 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The cell lysate was boiled for 5 min in 1 x SDS sample buffer (0.045 M Tris-HCl pH 6.8, 10% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue), containing 0.05 M DTT.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Mycoplasma free

    Yes
  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Constituent: 100% SDS Sample Buffer
  • Concentration information loading...
  • Lysate notes

    A549 cell lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The cell lysate was boiled for 5 minutes in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol.
  • Research areas

    • Kits/ Lysates/ Other
    • Lysates
    • Whole Cell Lysates
    • Human
    • Other
  • Background

    A-549 cells are used as an in vitro model for a type II pulmonary epithelial cell model for drug metabolism. Cells are cytokeratin positive by immunoperoxidase staining. This line was initiated in 1972 by D.J. Giard, et al. through explant culture of lung carcinomatous tissue from a 58-year-old Caucasian male (PubMed 4357758). Further studies by M. Lieber, et al. revealed that A549 cells could synthesize lecithin with a high percentage of desaturated fatty acids utilizing the cytidine diphosphocholine pathway (PubMed 175022).

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab7910 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
Use at an assay dependent concentration.
A549 cell lysate is ready to load on SDS-PAGE for Western blotting, 20 µg per lane is recommended for mini gel.
Notes
WB
Use at an assay dependent concentration.

A549 cell lysate is ready to load on SDS-PAGE for Western blotting, 20 µg per lane is recommended for mini gel.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1)

Publishing research using ab7910? Please let us know so that we can cite the reference in this datasheet.

ab7910 has been referenced in 1 publication.

  • Yougbare I  et al. NCS 613, a Potent PDE4 Inhibitor, Displays Anti-Inflammatory and Anti-Proliferative Properties on A549 Lung Epithelial Cells and Human Lung Adenocarcinoma Explants. Front Pharmacol 11:1266 (2020). PubMed: 32973507

Customer reviews and Q&As

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Question

I purchased this antibody and the A549 cell line as a positive control, but it's not working in WB. A different antibody worked to detect the protein, but not the control

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Abcam community

Verified customer

Asked on Nov 29 2012

Answer

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

Abcam Scientific Support

Answered on Nov 29 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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