Immunohistochemistry (Frozen sections) abreview for Anti-NG2 [LHM 2] antibody

Average
Abreviews
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (skin)
Specification
skin
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 3%

Other product details

Dilution
1/200
Incubation time
1 hour(s) and 30 minute(s)

Secondary antibody

Name
Non-Abcam antibody was used: anti-mouse F(ab)2
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488

Additional data

Additional Notes
The antibody works best with Aceton fixation (10 min 4°C Aceton) or Ethanol fixation (10 min 100% Ethanol, -20 °C).
Antibody works on frozen sections.
Antibody shows improved staining in the presence of Calcium. Therefore, BSA diluted in PBS+ (with Mg2+ and Ca2+) was used for blocking and antibody
dilution. PBS+ was also used for washing steps before application for primary antibody.

Staining intensity is good, but antibody shows relatively high background staining (bright dots) which were only a little bit improved when diluting higher than 1:200, however, staining intensity is then reduced. I assume that this is a problem of the particular batch.
Antibody works well with the following incubation times:
1h 30min at RT
4°C over night
Abcam response
Thank you for your feedback, I am sure this information will be helpful to other researchers. We are currently looking into why this antibody caused the high background mentioned in the additional notes.

Dr. Sonja Muffler

Verified customer

Submitted Dec 14 2006

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