Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hiPSC-RPE)
Permeabilization
Yes - 0.1% tritonX100
Specification
hiPSC-RPE
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 0.05%tritonX/5%NDS
Dilution
1/50
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Alexa fluor anti-mouse 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
After blocking for 1hr at RT in 10%NDS/0.1%tritonX, human iPSC-RPE (cryo sections) were incubated with TIMP3 (1:50 dilution) overnight at 4C. The next day, sections were washed and incubated with anti-mouse-488 secondary (1:500) for 1hr at RT, washed, and mounted.
Image was obtained at 40x mag using a confocal microscope. TIMP3 (in the green channel) should be basal in the ECM. EZRIN, an apical protein, is in the red channel for reference. We would not recommend this antibody for IF on human cells because the signal from the TIMP3 antibody is not properly localized.
Abcam response
We would like to thank this researcher for submitting their review. This antibody had not been tested in ICC and we were unsure of what the results would be. The reviewer received an AbTrial discount code for using the antibody in an untested application.
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.
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提交于 May 14 2018