Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Rat ovary, uterus)
Specification
Rat ovary, uterus
Fixative
10% buffered formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 20°C
Other product details
Dilution
1/50
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS (with Tween)
Secondary antibody
Name
Non-Abcam antibody was used: Dako Envision (Rabbit)
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Additional data
Additional Notes
High temperature antigen retrieval, using the Milestone RHS-2 microwave with pressure vessel at 110C for 2 minutes, produced the best nuclear staining.
Other pretreatments were also tested eg 100C for 10 minutes, using either 10mM citrate (pH 6.0) or 1mM EDTA buffer (pH 8.0). These produced positive, but less intense, nuclear staining.
This technique produced a similar staining pattern and intensity when the primary antibody was applied at 1:50 for 2 hours at room temperature.
Other pretreatments were also tested eg 100C for 10 minutes, using either 10mM citrate (pH 6.0) or 1mM EDTA buffer (pH 8.0). These produced positive, but less intense, nuclear staining.
This technique produced a similar staining pattern and intensity when the primary antibody was applied at 1:50 for 2 hours at room temperature.
Abcam response
Abcam recommends overnight incubation of the primary antibody at 4C to increase signal.
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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投稿 Nov 27 2007