Immunocytochemistry/ Immunofluorescence abreview for Anti-LXR alpha antibody

Below Average
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa transiently expressing human LXRa.)
Specification
HeLa transiently expressing human LXRa.
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TritonX-100 in PBS for 10 min at 4'C
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 23°C

Other product details

Dilution
1/200
Incubation time
30 minute(s) · Temperature: 37°C · Diluent: PBS with 0.1% BSA

Secondary antibody

Name
Non-Abcam antibody was used: Goat anti rabbit IgG with Alexa-568 from Invitroge
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 568
Dilution
1/500

Additional data

Additional Notes
This Ab recognized an overexpressed LXRa in nucleus (upper right corner of the picture), not endogenous LXRa (other cells) with strong background signal. Longer blocking might have reduced a background. Not suitable for immunostaining with currrent protocol.
Abcam response
Thank you for you honest and thoughtful feedback. Based on a previous Abreview, I would recommend a longer incubation time with the primary (1 hour at RT), at a higher concentration 1/100. I would also recommend diluting the primary antibody in TBS + 3% dried milk + 0.1% Triton X-100, which can also be used as the block. This antibody does appear to need as longer blocking time, as the previous reviewer blocked for 20 hours.

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Submitted May 21 2008

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