Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody

Excellent
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse ES cell (YFP) reporter line.)
Permeabilization
Yes - 5% donkey serum in PBST (PBS / 0.1% v/v Triton X-100 (Sigma))
Specification
Mouse ES cell (YFP) reporter line.
Blocking step
5% donkey serum in PBST (PBS / 0.1% v/v Triton X-100 (Sigma)) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde

Other product details

Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% donkey serum in PBST
Dilution
1/2000

Secondary antibody

Dilution
1/1000
Name
Non-Abcam antibody was used: Donkey anti-Rabbit Alexa Fluor 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488

Additional data

Additional Notes
Image shows ESC reporter line where Venus is expressed as a fusion construct with the transcription factor. Red field: IHC-Fr for transcription factor (Alexa 564). Green field: IHC-Fr with ant-GFP (ab6556). Blue field: DAPI. We tried many GFP antibodies in the past, but few actually reliably cross-react reliably with Venus. We tested the antibody on two different Venus(YFP) expressing ES cell lines: one driven by the endogenous promoter (Transcription Factor promoter-Venus, cytoplasmic Venus), the other as a fusion protein (Transcription Factor-Venus, nuclear Venus). The antibody reliably identified Venus in both cases.

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Submitted Sep 21 2015

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