E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395)
- Datasheet
- Specific References (2)
- Protocols
Overview
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Product nameE. coli Positive Control (Escherichia coli ) Whole Cell Lysate
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General notes
This E.coli lysate contains a 41 kd recombinant protein expressing 12 commonly used Tags which are fused to the N-terminus of the protein in the order of:
T7 HSV
C-Myc
VSV-g
Glu-Glu
V5
E-tag
FLAG (DDDDK)
S-tag
HA
6x HisGST
Tag sequences: T7 = CMASMTGGQQMG
HSV = QPELAPEDPEDC
CMYC = EQKLISEEDLC
VSV-g = YTDIEMNRLGK
GluGlu = EYMPME
V5 = GKPIPNPLLGLDST
E-tag = GAPVPYPDPLEPR
FLAG = DYKDDDDK
HA = YPYDVPDYAC
6xHIS = HHHHHHExcellent as a loading control for Western blotting. Lysate expresses 11 different epitope tags.
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Tested applicationsSuitable for: WBmore details
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Concentration information loading... -
PurityWhole Cell Lysate
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Purification notesE. coli were lysed with SDS-PAGE sample loading dye, boiled at 95°C for 10 minutes, then centrifuged at 13,000 g for 15 minutes.
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Lysate notes
Excellent as a loading control for Western blotting. Lysate expresses 11 different epitope tags.
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Research areas
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BackgroundEscherichia coli is a gram negative bacillus that belongs to a larger group of Enterobacteriae - bacteria that inhabit the gastrointestinal tract. Although usually a harmless resident of the gut, some strains have the potential to cause serious problems, especially where there is an immature immune system or immunosuppression, or where the subtype of organism has acquired the ability to produce pathogenic toxins.
Applications
Our Abpromise guarantee covers the use of ab5395 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. Use 0.5 - 2.0 µl for 10 - 12% gels. |
Images
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Western blot - E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395)All lanes : Anti-6X His tag® antibody - ChIP Grade (ab9108) at 1/1000 dilution
Lane 1 :E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 2 µl
Lane 2 :E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 1 µl
Lane 3 :E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 0.5 µl
Lane 4 :E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 0.25 µl -
Western blotAnti-Myc tag antibody (ab107247) at 1 µg/ml +E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 10 µg
Secondary
Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 41 kDa (why is the actual band size different from the predicted?)
This WB was performed using an E. coli lysate that expresses a 41kDa recombinant protein comprised of 11 commonly used epitope tags (T7, HSV, C-Myc, VSV-g, Glu-Glu, V5, E-tag, FLAG, S-tag, HA and 6xHis). This lysate is available as ab5395.
References
This product has been referenced in:
- Májek P et al. Improved Coomassie Blue Dye-Based Fast Staining Protocol for Proteins Separated by SDS-PAGE. PLoS One 8:e81696 (2013). Read more (PubMed: 24278455) »
- Miriam_Sans, Itzel_Vargas-Prez, Luis_Quintales, Fr Gcn5 facilitates Pol II progression, rather than recruitment to nucleosome-depleted stress promoters, in Schizosaccharomyces pombe. Nucleic Acids Res : 1-11 (2011). WB ; Schizosaccharomyces pombe . Read more (PubMed: 21515633) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"