Application
Western blot
Sample
Hamster Cell lysate - whole cell (Chinese Hamster Ovary cell line)
Loading amount
50 µg
Specification
Chinese Hamster Ovary cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s)
Secondary antibody
Name
Non-Abcam antibody was used: Pierce goat anti-rabbit (H+L)
Host species: Goat
Clonality: Polyclonal
Conjugation: Alkaline Phosphatase
Host species: Goat
Clonality: Polyclonal
Conjugation: Alkaline Phosphatase
Detection
Detection method
AP
Exposure
2 minute(s) and 0 second(s)
Bands
Specific: 110 kDa
Positive control
Whole cell lysate of human cell line known to contain antigen for GRP94
Additional data
Additional Notes
Background can be a problem, lower dilutions of the primary may help. Using alkaline phospatase secondary cleaned up image significantly.
10% reducing gel, transferred to PVDF membrane, incubated primary overnight rocking at 4C, blocked with 4% skim milk, 5X 5min wash with TBS-T, AP conjugates secondary incubated 1 hour at room temperature, 5X 5min wash with TBS-T, visualized with BCIP/NBT reagents. This antibody is not a good marker for biochemical fractionation of the cell. Shows in all fractions.
10% reducing gel, transferred to PVDF membrane, incubated primary overnight rocking at 4C, blocked with 4% skim milk, 5X 5min wash with TBS-T, AP conjugates secondary incubated 1 hour at room temperature, 5X 5min wash with TBS-T, visualized with BCIP/NBT reagents. This antibody is not a good marker for biochemical fractionation of the cell. Shows in all fractions.
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提交于 Aug 11 2006