Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Specification
Kidney
Fixative
10% neutral buffered formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH 6.0 (20 min, 93*C)
Permeabilization
No
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 21°C
Other product details
Concentration
8.5 µg/ml
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 21°C · Diluent: 0.3% Triton X-100 in PBS
Secondary antibody
Name
Non-Abcam antibody was used: Vector Labs Goat Anti-Rabbit IgG Antibody
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Concentration
7 µg/ml
Additional data
Additional Notes
Peroxidase Block: 10 min methanol/H2O2
ABC: 30 min Vectastain ABC (2%)
Peroxidase substrate: 5 min Vector NovaRED
Nuclear stain: 10 sec Vector Hematoxylin QS
Notes:
-Further dilution below 8.5 ug/ml not recommended for this application.
-Heat-mediated antigen retrieval necessary, no retrieval or Trypsin-CaCl2 do not produce positive stain. Citrate buffer produced better results than Tris-EDTA.
Image: A = ApoB kidney tubules @ 8.5 ug/ml, B = ApoB kidney glomerulus @ 8.5 ug/ml
ABC: 30 min Vectastain ABC (2%)
Peroxidase substrate: 5 min Vector NovaRED
Nuclear stain: 10 sec Vector Hematoxylin QS
Notes:
-Further dilution below 8.5 ug/ml not recommended for this application.
-Heat-mediated antigen retrieval necessary, no retrieval or Trypsin-CaCl2 do not produce positive stain. Citrate buffer produced better results than Tris-EDTA.
Image: A = ApoB kidney tubules @ 8.5 ug/ml, B = ApoB kidney glomerulus @ 8.5 ug/ml
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投稿 Jan 24 2013