Immunohistochemistry (Frozen sections) abreview for Anti-GFP antibody [LGB-1]

Poor
Abreviews
Application
Immunohistochemistry (Frozen sections)
Blocking step
NOVOCASTRA protein block as blocking agent as blocking agent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Sample
Mouse Tissue sections (E10.5 mouse embryo)
Specification
E10.5 mouse embryo
Permeabilization
Yes - PBS / 0.5% v/v Triton X-100 (Sigma)
Fixative
Paraformaldehyde

Other product details

Incubation time
12 hour(s) and 0 minute(s) · Temperature: 25°C
Dilution
1/200

Secondary antibody

Name
Non-Abcam antibody was used: donkey anti-mouse 568 (Invitrogen)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor­ 568
Dilution
1/1000

Additional data

Additional Notes
Embryos were electroporated with Plasmid 13776: pCAG-Cre:GFP from Addgene and then fixed with 4% PFA overnight at 4 degree. Samples were embedded in gelatin and sectioned using a cryostat. Left image is DAPI plus GFP (without staining). Right images is using anti-GFP antibody [LGB-1] (ab291) at 1:200 dilution, the secondary antibody used was donkey anti-mouse Alexa 568 from invitrogen (1:1000). GFP can be detected using anti-GFP ab13970, but not anti-GFP ab291.
Abcam response
Thank you for sharing this data with us. Previously ab291 has not been tested in IHC-Fr and based on this abreview, it looks like this antibody is not suitable for IHC-Fr under these conditions. Abcam would welcome feedback from other customers currently using this product.

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Submitted Mar 20 2014

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