Western blot abreview for Anti-B Raf antibody [EP152Y]

Inconclusive
Abreviews
Application
Western blot
Sample
Mouse Tissue lysate - whole (brain, liver, skin from Tg animal expressing Human)
Loading amount
100 µg
Specification
brain, liver, skin from Tg animal expressing Human
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Other product details

Dilution
1/2000
Incubation time
16 hour(s) and 0 minute(s)

Secondary antibody

Name
Non-Abcam antibody was used: Goat anti rabbit HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL
Exposure
5 minute(s) and 0 second(s)
Bands
Specific: 95 kDa
Positive control
This antibody data sheet suggests it does not cross react with Mouse BRAF. But I ran three Wb using Human melanoma, Hela Cell line lysates and also used Wild Type mouse melanocytes, p53 -/- MEFs and various tissue lysates from mouse such as BRAIN, Liver, Lung, Kidney, Heart,and skin (from Transgenic mouse which has human BRAF). But this antibody reacts with both human and mouse BRAF. With Human it showed one band and for mouse it showed two bands because mouse BRAF undergoes alternate splicing.
Negative control
Negative control various mouse lysate as mention above.

Additional data

Additional Notes
This anitbody reacts human and mouse very well infact mouse showed better signal than human
Abcam response
Thank you for your Abreview. Previous data suggested that the antibody did not cross-react with mouse. I am sure this information will be helpful to future customers.

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Submitted Oct 02 2006

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