Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (BT474 Xenografts)
Specification
BT474 Xenografts
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
Normal Goat Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 21°C
Other product details
Dilution
1/50
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 21°C · Diluent: 0.1% TBS Tween20
Secondary antibody
Name
Non-Abcam antibody was used: Dako Mouse Envision
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Additional data
Additional Notes
Image is of BT474 xenograft with Primary antibody @ 1:50 with antigen retrival @ 110C for 2 minutes in Citrate buffer.
Also tested @ 1:100 and 1:200 with no pretreatments and with antigen retrival @ 110c for 2 minutes in EDTA buffer.
Weak nuclear staining with background wash.
Also tested several other xenografts where no staining was observed.
Also tested @ 1:100 and 1:200 with no pretreatments and with antigen retrival @ 110c for 2 minutes in EDTA buffer.
Weak nuclear staining with background wash.
Also tested several other xenografts where no staining was observed.
Abcam response
We are thankful for the feedback and are sorry to hear that the customer obtained weak staining.
Option would have been to try longer antigen retrieval times including lower temperature (<110C). However, as the customer used an automated stainer, longer than 2 minutes retrieval resulted in tissue damage.
Option would have been to try longer antigen retrieval times including lower temperature (<110C). However, as the customer used an automated stainer, longer than 2 minutes retrieval resulted in tissue damage.
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.
Abcam user community
Verified customer
提交于 Mar 17 2010