Immunocytochemistry/ Immunofluorescence abreview for Anti-Giantin antibody [9B6] - Golgi Marker

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Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Muscle cell line (C2C12))
Permeabilization
Yes - Triton in PBS
Specification
Muscle cell line (C2C12)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1000µg/mL · Temperature: 21°C
Fixative
Paraformaldehyde

Other product details

Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1mg/ml BSA in PBS
Dilution
1/200

Secondary antibody

Name
Non-Abcam antibody was used: AlexaFluor 555
Host species: Goat
Clonality: Monoclonal
Conjugation: Alexa Fluor® 555

Additional data

Additional Notes
Our purpose for using this antibody was for co-localization studies with the Golgi. However, we found that the antibody labelled the nucleus at a high dilution (1:20) and very little staining characteristic of the golgi was seen at either a high or low dilution. There may have been a very small subset of cells (1 in 300-400) that had possible perinuclear staining (which was predicted from other studies that used a different Giantin antibody in myoblasts). Upon inquiring with Abcam Technical Support, I was told that the portion of the protein this antibody was raised against has low homology between human and mouse, which likely explains the lack of golgi staining and off-target nuclear staining. I used 4% PFA fix, Triton permeabilization, BSA block, and AlexaFluor secondary.
Abcam response
Thank you for submitting an Abreview and you valuable feedback about the use of ab37266 in mouse. Based on the alignment of the immunogen and the mouse Golgb1 protein and your data, the antibody is most likely not suitable for mouse.

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Submitted Mar 01 2016

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