Primary antibody:
Invitrogen PA5-42693 (Rabbit)
Source material:
Not specified
Secondary antibody:
Cell Signaling 7074S (Goat)
Lysis Conditions:
RIPA buffer
at
22°C
for
1 min
Gel Conditions
Run conditions:
200 V / 0 Amps for 0.5 hr
Reducing conditions:
β-mercaptoethanol
Reducing-HepG2 lysate; boiled for 10 mins at 90 degrees Celsius
Transfer Conditions
Membrane type:
PVDF
Transfer method:
Wet
Transfer buffer:
Tris-glycine with 10% methanol
Transfer voltage:
100 V
Transfer current:
0 Amps
Transfer temperature:
4°C
Transfer duration:
1 hr
Blocking
Blocking buffer:
Superblock
Blocking temperature:
22°C
Duration:
1 hr
Primary Incubation
Antibody 1 dilution
1:1000
Incubation temperature
22°C
Incubation duration
1 hr
Number of washes
3
Wash buffer
TBST
Wash duration
5 min
Secondary Incubation
Antibody 2 dilution
1:10
Incubation temperature
22°C
Incubation duration
1 hr
Number of washes
3
Wash buffer
TBST
Wash duration
5 min
Detection
Detection method
Enhanced chemiluminescence (ECL)
Expected band size
44 kDa
Result:
No specific bands
Background:
No background
Related Publications
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Record 3176 author: Amber Morey
Last edit: Thu Feb 23, 2023
Extremely faint band at appropriate size - potentially need more protein per lane