Target protein:
Ferroportin
Primary antibody:
Santa Cruz Biotech sc-518125 (Mouse)
Source material:
Tissue: Placental explants
Placenta from a normal gestation (third trimester of pregnancy).
Secondary antibody:
Sigma-Aldrich 12-349 (Goat)
Lysis Conditions:
RIPA buffer
at
4°C
for
15 min
Gel Conditions
Minigel, Any kD % acrylamide
Run conditions:
200 V / 0 Amps for 30 min
Reducing conditions:
Dithiothreitol
Lane 1: Placental explant supernatant
Lane 2: Placental lysate
Transfer Conditions
Membrane type:
PVDF
Transfer method:
Semi-dry
Transfer buffer:
Tris-glycine with 20% methanol
Transfer voltage:
25 V
Transfer current:
1.0 Amps
Transfer temperature:
22°C
Transfer duration:
0.5 hr
Blocking
Blocking buffer:
2% milk in TBST. Incubation system: Manual (blocking and primary ab) & SNAP id (washes and secondary ab)
Blocking temperature:
22°C
Duration:
0.5 hr
Primary Incubation
Antibody 1 dilution
1:200
Incubation temperature
4°C
Incubation duration
16 hr
Number of washes
3
Wash buffer
TBST, 0.1% Tween-20
Wash duration
5 min
Secondary Incubation
Antibody 2 dilution
1:20,000
Incubation temperature
22°C
Incubation duration
0.16 hr
Number of washes
4
Wash buffer
TBST, 0.1% Tween-20
Wash duration
0 min
Detection
Detection method
Enhanced chemiluminescence (ECL)
Expected band size
62 kDa
Result:
Weak single band
Background:
Multiple non-specific bands
Related Publications
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Record 10015 author: Luiza Perucci
Last edit: Fri Dec 1, 2023
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