Immunocytochemistry/ Immunofluorescence abreview for Anti-HA tag antibody - ChIP Grade

Average
Abreviews
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Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.1% Triton X-100/0.02% SDS
Specification
HeLa
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
3% paraformaldehyde, 0.02% glutaraldehyde

Other product details

Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: 1% BSA, 0.1% Triton, 0.02% SDS in PBS
Dilution
1/500

Secondary antibody

Dilution
1/500
Name
Non-Abcam antibody was used: goat anti rabbit IgG, Alexa Fluor 647
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor­ 647

Additional data

Additional Notes
The antibody works very well for the detection of cytoplasmic proteins by IF. The antibody produces a nuclear background, however, and is thus not suitable for the detection of low abundance nuclear proteins. In the figure, a non-Abcam mouse anti-HA and ab9110 are used to detect a low-abundance HA-tagged nuclear envelope protein. Nuclear rim staining is clearly discernable with the mouse anti-HA antibody (upper panel). Nuclear rim staining of the protein of interest is barely recognizable with ab9110 because of a significant nuclear background produced by this antibody. This background is also visible in non-transfected cells (asterisks).

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Submitted Jan 25 2017

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