Western blot abreview for Anti-Histone H3 (citrulline R2) antibody [EPR17703]

Excellent
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HL60 cell line)
Gel Running Conditions
Non-reduced Denaturing (4-20% gradient gel)
Loading amount
40 µg
Treatment
Differentiated in 1.25% DMSO for 5 days, treated with 4uM calcium ionophore A23187 for 15 mins at 37C.
Specification
HL60 cell line
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Other product details

Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA in TBS-Tween (0.1%)
Dilution
1/2000

Secondary antibody

Dilution
1/5000
Name

Detection

Detection method
SuperSignal™ West Pico Chemiluminescent
Negative control
Lane 2 - Undifferentiated HL60 cell line and Lane 3 - HL60 cell line, differentiated in 1.25% DMSO for 5 days but not treated with calcium ionophore.
Exposure
20 second(s)
Bands
Specific: 17 kDa
Positive control
Lane 4 - HL60 cell line, differentiated in 1.25% DMSO for 5 days, then treated with 4uM calcium ionophore A23187 for 15 mins at 37C.

Additional data

Additional Notes
Lane 1: Size marker. Lane 2: Undifferentiated HL60 cells. Lane 3: HL60 cells, differentiated in 1.25% DMSO for 5 days. Lane 4: HL60 cells, differentiated in 1.25% DMSO for 5 days, then treated with 4uM calcium ionophore A23187 for 15 mins at 37C. No cross-reactivity with unmodified H3 was observed, even at higher exposures.

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Submitted Aug 12 2016

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