Application
Immunocytochemistry/ Immunofluorescence
Sample
Monodelphis domestica Cell (testis)
Permeabilization
Yes - 0.5% Triton X-100
Specification
testis
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Paraformaldehyde
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 10% goat serum in TBST
Dilution
1/50
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: donkey anti mouse Alexa Fluor 488 (A21202)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
Used abtrial program to test antibody, thank you!
Paraformaldehyde was used at 4% concentration. Used no primary control as well (right panel), background fluorescence in both with no staining in spermatogonial stem cells (left panel, 1:50 dilution) as would be expected. Blue = DAPI.
Also tried MeOH fix, longer blocking times, higher and lower concentrations of triton x-100 (from none to 1%), lower dilutions of primary (1:100, 1:150), and another lot of secondary antibody. Used samples from 2 month, 8 month, and 3 year old opossums (images are from 3 year old, testis). Seems to just not be compatible with Monodelphis testis despite high homology, will test once more when new samples are collected.
Abcam response
Thank you so much for testing this antibody in opossum. We appreciate very much your efforts, but are sorry to hear that the antibody did not work well in this species.
The customer received a discount on a future antibody purchase for testing this product. We welcome additional feedback from other customers as well.
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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Verified customer
提交于 Sep 14 2016