Western blot abreview for Anti-KDM5C / Jarid1C / SMCX antibody

Inconclusive
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (different cell lines)
Gel Running Conditions
Reduced Denaturing (7,5% gel)
Loading amount
50 µg
Specification
different cell lines
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Other product details

Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 2.5%milk/PBS-tween 0.05%
Dilution
1/250

Secondary antibody

Dilution
1/8000
Name
Non-Abcam antibody was used: Biorad
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
CLARITY (Biorad)
Exposure
1 hour(s), 0 minute(s) and 0 second(s)
Bands
Specific: 170 kDa Non-specific: several kDa
Positive control
We used different cell lines, total extracts (nuclear + cytosolic), that should be positive for JARID1c.

Additional data

Additional Notes
Several unspecific bands, very difficult to detect the protein. We get a band around the expected molecular weight but it is difficult to trust since we have dozens of other bands throughout the membrane. if we use more diluted ab, no bands are detected. We used different cell lines, ab dilutions, exposure times etc… membrane: nitrocellulose lysis buffer: RIPA + protease inhibitors (Roche)
Abcam response
We appreciate the valuable data provided in this Abreview. Since the antibody didn't work as it was supposed to in our previously tested and guaranteed species and application, we have followed up with the customer in accordance to our Abpromise (www.abcam.com/abpromise) and issued a replacement.

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Verified customer

Submitted Nov 29 2013

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