Western blot abreview for Anti-WDR5 antibody

Good
Abreviews
This product is known to not work in this application or species.
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (nuclear)
Loading amount
10 µg
Specification
nuclear
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C

Other product details

Dilution
1/500
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBSTween-20 0.1% and same with 2% BSA

Secondary antibody

Name
Non-Abcam antibody was used: goat anti rabbit IgG (H&L) HRPO
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/20000

Detection

Detection method
Pierce Supersignal
Exposure
5 minute(s) and 0 second(s)
Positive control
HEK293 whole cell lysate
A431NS carcinoma whole cell lysate
Negative control
N/A

Additional data

Additional Notes
Reacts with human cell lines. Does not react with Drosophila nuclear lysate despite 100% homology with peptide use to generate this antibody and the Drosophila version of this protein (wds). 50 mg of nuclei were isolated from adults and lysed in 400 mM KCl, 10 mM HEPES, 0.05% Igepal-630, 2mM DTT, 10% glycerol at pH 7.6 or RIPA buffer (both supplemented with protease inhibitors) followed by 5 10 second pulses of sonication at low power.
Lane 1: Drosophila high salt extraction (nuclear)
Lane 2: Drosophila RIPA extraction (nuclear)
Lane 3: HEK 293 whole cell lysate
Lane 4: A431NS carcinoma whole cell lysate

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Submitted Aug 02 2010

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