Western blot abreview for Anti-Histone H3 (tri methyl K9) antibody [EPR16601] - ChIP Grade

Below Average
Abreviews
Application
Western blot
Sample
Human Cell lysate - nuclear (Melanoma)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
3000 cells
Specification
Melanoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Other product details

Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/1000

Secondary antibody

Dilution
1/5000
Name
Non-Abcam antibody was used: Goat Anti-Rabbit IgG Antibody
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL+
Negative control
Not used.
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 15 kDa Non-specific: 12, 32, 80, 130 kDa
Positive control
Not used.

Additional data

Additional Notes
Lane 1 - 501 mel (human derived melanoma cell line). Lane 2 - SKmel147 (human derived melanoma cell line). 15 kDa is H3, as would be expected in both lanes. There are multiple distinct nonspecific bands in both lanes. I also tried 1:10,000 dilution, but saw no difference in background.
Abcam response
Thank you for sharing this data with us. ab176916 has previously been tested in western blot on HeLa and NIH3T3 whole cell lysates (please see our western blot image on the datasheet). However, based on the data shown here, it seems that not all protocols are equally suitable for ab176916, in western blotting. We would recommend trying this antibody at a much highly dilution >1:20,000 under your conditions. We welcome feedback from other researchers currently using this antibody.

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Submitted Dec 23 2016

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