Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (stomach)
Specification
stomach
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA (pH 9.0)
Permeabilization
No
Blocking step
RE7102, NovoLink Polymer Detection System as blocking agent for 5 minute(s) · Concentration: 1% · Temperature: RT°C
Other product details
Dilution
1/1000
Incubation time
1 hour(s) and 0 minute(s)
Secondary antibody
Secondary antibody
None used
Additional data
Additional Notes
ab40278 reacts well with lymphocytes, but it cannot detect the RUNX3 in normal gastric or colonic epithelial cells.
The retrieving methods had been ever tried including retrieved by autoclave at operating temperature in Tris-EDTA (pH 9.0) solution for 1.5 minutes or in 0.01 M citrate buffer (pH 6.0) for 1.5 minutes, retrieved by microwave oven at operating temperature in 0.01 M citrate buffer (pH 6.0) for 5 minutes, incubated in 0.01% trypsin for 15 minutes, incubated in 0.01% pepsin for 15 minutes and no retrieving.
The incubation time for primary antibody performed either for 1 hour at room temperature or overnight at 4c.
Among these retrieval methods, autoclave at operating temperature in Tris-EDTA (pH 9.0) solution for 1.5 minutes is the best. RUNX3 only expressed in lymphocytes, however neither expressed in cytoplasm nor nucleus of gastric epithelial cells.
For each retrieval conditions, only one staining process had been carried out.
Since the staining of lymphocytes was satisfied by using 1ug/ml antibody dilutions, no other dilutions had been carried out.
The retrieving methods had been ever tried including retrieved by autoclave at operating temperature in Tris-EDTA (pH 9.0) solution for 1.5 minutes or in 0.01 M citrate buffer (pH 6.0) for 1.5 minutes, retrieved by microwave oven at operating temperature in 0.01 M citrate buffer (pH 6.0) for 5 minutes, incubated in 0.01% trypsin for 15 minutes, incubated in 0.01% pepsin for 15 minutes and no retrieving.
The incubation time for primary antibody performed either for 1 hour at room temperature or overnight at 4c.
Among these retrieval methods, autoclave at operating temperature in Tris-EDTA (pH 9.0) solution for 1.5 minutes is the best. RUNX3 only expressed in lymphocytes, however neither expressed in cytoplasm nor nucleus of gastric epithelial cells.
For each retrieval conditions, only one staining process had been carried out.
Since the staining of lymphocytes was satisfied by using 1ug/ml antibody dilutions, no other dilutions had been carried out.
Abcam response
We appreciate customers detailed feedback and are sorry to hear that the antibody did not work in customer's hands as wished.
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.
Dr. Yuan LI
Verified customer
投稿 Aug 20 2010