Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Bone and Aortic Valve Tissue)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Trypsin-CaCl 0.001mg/mL 30 min 37*C
Permeabilization
No
Specification
Bone and Aortic Valve Tissue
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
10% NBF
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 21°C · Diluent: PBS/0.3%TritonX-100
Concentration
20 µg/ml
Secondary antibody
Dilution
1/50
Name
Non-Abcam antibody was used: Vector Labs VectaStain Universal Elite ABC Kit
Host species: Horse
Clonality: Polyclonal
Conjugation: Biotin
Host species: Horse
Clonality: Polyclonal
Conjugation: Biotin
Additional data
Additional Notes
Fixative: 10% NBF 2 days RT
EDTA decalcification (bone)
Peroxidase Block: Methanol/H2O2 10 min
Color Substrate: Vector NovaRED (5 min)
Counterstain: Vector Hematoxylin QS (10 sec)
Notes:
This antibody only got very slight staining on marrow cells at high concentration and would start to lightly non-specifically stain marrow components.
Fluorescence was also done:
- Tris/EDTA pH 9.0, 1 hour with solution cooling from 85 – 55 *C followed by removal of bath to room temperature for 20 min
- primary: 40-10ug/mL secondary: Alexa Fluor 568 goat anti-mouse IgG
- This only resulted in very slight staining in bone marrow cells at 40ug/mL.
- the valve was completely clean with no background staining with both methods
Image: Bone Marrow A) NovaVector Red, primary 20ug/mL, B) Fluorescence 40ug/mL
Abcam response
Thank you for your review of this untested species. We appreciate the time and effort you spent testing this antibody in pig. You may improve your signal by keeping the concentration of the primary at 10 ug/mL and increasing the incubation to overnight at 4 degrees.
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
投稿 Jan 24 2013