Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (LDT brain slices)
Specification
LDT brain slices
Permeabilization
Yes - TritonX
Fixative
PFA/Gluteraldehyde
Other product details
Incubation time
24 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: PBS-Triton X- goat serum
Dilution
1/400
Secondary antibody
Name
Abcam antibody:Goat Anti-Guinea pig IgG H&L (Alexa Fluor 594)
Dilution
1/500
Additional data
Additional Notes
Bad background staining and nonspecific staining.
I have spoken with the Abcam team and will try reducing the concentration and incubation times in future trials to obtain better results.
Abcam response
Thank you for your valuable feedback. Abcam have spoken to the customer and have suggested the following protocol tips:
1. Use the recommended 1/1000 dilution rather than 1/400.
2. If you are going to incubate overnight, do this at 4¹C rather than at 25¹C.
At higher temperatures, the antibody is more likely to bind unspecifically. Lowering the incubation time will therefore also help reduce your background.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Jason Teem
Verified customer
提交于 Sep 10 2014