Q: In NB800-PC6, were the 293 cells transfected with WT1 expression plasmids? We have 293 cell lysates and it showed negative WT1 expression. Just want to make sure if our antibody is not working.

A: This protein lysate was derived from human HEK293 cells (without any transfection) and it is a whole cell lysate. WT1's mRNA is expressed significantly in HEK294 cells protein atlas site and given the fact that our Wilms Tumor 1 antibody (6F-H2) [NB110-60011] showed positive bands on HEK293 cells lysate, you should be good for considering this lysate as a positive control for WT1.

293 Whole Cell Lysate

Novus Biologicals | Catalog # NB800-PC6

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Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

SDS-PAGE, Western Blot

Product Specifications

Application Notes

Ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOuT dissociating agents. Heat lysate - 95C for 5 minutes and rapidly cool. If dissociating conditions are desired, add reducing agent prior - heating. The recommended loading volume per lane is 10-20 ul depending on the size format of your gel.

Specificity

293 whole cell extract lysate

Type

Cell

Subcellular Fraction

Whole

Scientific Data Images for 293 Whole Cell Lysate

Western Blot: 293 Whole Cell Lysate [NB800-PC6]

Western Blot: 293 Whole Cell Lysate [NB800-PC6] - Lane 1: Coomassie stain of 0.02 ml 293 extract (NB 800-PC6), separated in a 4-20% gradient gel under reducing conditions. Lane 2: Molecular weight standard.

SDS-PAGE: 293 Whole Cell Lysate [NB800-PC6]

SDS-Page: 293 Whole Cell Lysate [NB800-PC6] - Coommassie stained SDS-PAGE of 20 ug of (1) Hela WCL, (2) Jurkat WCL, (3) HEK 293 WCL, (4) MCF-7 WCL, (5) A 549 WCL, (6) Raji WCL separated using a 4-20% gradient gel under reducing conditions. Molecular weight standards are shown.

Formulation, Preparation, and Storage

Formulation

Standard RIPA buffer including protease and phosphatase inhibitors and 0.1% SDS diluted in 2x Laemmli Sample Buffer (125 mM Tris HCl, 4% SDS, 50% Glycerol and 0.02% bromophenol blue, pH 6.8)

Concentration

2.0 mg/ml

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: 293

This ready-to-use whole cell lysate is derived from a cell line or tissue using highly refined extraction protocols to ensure exceptionally high quality, protein integrity and lot-to-lot reproducibility. All extracts are tested by SDS-PAGE using 4-20% gradient gels and immunoblot analysis using antibodies to key cell signaling components to confirm the presence of both high molecular weight and low molecular weight proteins.

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Product Documents for 293 Whole Cell Lysate

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Product Specific Notices for 293 Whole Cell Lysate

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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