VALIDATION IMAGES
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: (-): HeLa, (+): HeLa+ Human α-TNF
(20ng/ml, 30min)
Protein loading quantity: 20 μg
Exposure time: 30 s
Predicted MW: 36 kDa
Observed MW: 36 kDa
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: (-): C6, (+): C6+ Calyculin A (100nM,
30min)
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 36 kDa
Observed MW: 36 kDa
Tissue: (A) Rat kidney, (B) Rat kidney, then the tissue was incubated with alkaline phosphatase (50
U/ml) at 37℃ for 1 hour
Section type: Formalin-fixed & Paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:100
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal for bsm-60711R
Tissue: (A) Mouse kidney, (B) Mouse kidney, then the tissue was incubated with alkaline
phosphatase (50 U/ml) at 37℃ for 1 hour
Section type: Formalin-fixed & Paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:100
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to use)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal for bsm-60711R