Application
Western blot
Sample
Cat Cell lysate - whole cell (Skin Fibroblasts and Vaccination Associated Sarcom)
Gel Running Conditions
Reduced Denaturing (12% Gel)
Loading amount
10 µg
Specification
Skin Fibroblasts and Vaccination Associated Sarcom
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 22°C · Diluent: TBST
Concentration
10 µg/ml
Detection
Detection method
ECL-Select
Negative control
GAM-only control
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 0 kDa Non-specific: 25, 30, 35, 40, 45, 50, 55 kDa
Positive control
Human PC3 Cell Line
Additional data
Additional Notes
I followed the protocol suggested by your data sheet. I got only non-specific staining.
Image 1:
Lane 1: Cat Skin Fibroblasts
Lane 2: Cat VAS Cell Line
Lane 3: Human PC3 Cell Line
Lanes 1-3 were stained with GAM 1:10,000 only (prepared in TBST as suggested in the ab1316 data sheet).
Lane 4 ladder: 250, 150, 100, 75, 50, 37, 25 kDa.
Lane 5: Cat Skin Fibroblasts
Lane6: Cat VAS Cell Line
Lane 7: Human PC3 Cell Line
Lanes 5-7 were stained with 10 ug/mL ab1316 in TBST for 1 hour at room temp, after being blocked with 3% milk in TBST. GAM was used at 1:10000 in TBST for 1 hour. (as suggested by the data sheet)
Staining is non-specific in all samples.
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.
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提交于 Nov 06 2017