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Author Topic: VCAM mAb troubleshooting  (Read 5419 times)

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Offline lmk30

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VCAM mAb troubleshooting
« on: March 30, 2005, 06:39:28 AM »
Hi,
I've just started staining with mouse mAb's against human VCAM-1 and ICAM-1 (Santa Cruz) but have not had any luck... I'm using paraffin embedded human biopsies and the protocol(which is otherwise standard) includes citrate antigen retrieval, blocking in 2% secondary Ab species serum for 1h @RT, incubation with the primary Ab overnight @4 deg. C at dilutions ranging from 1:50 to 1:100. I have also tried DAB times up to 10 min. The above protocol works very well normally but for these antibodies I just can't seem to get any staining. Maybe I'm just thick but if anyone has any suggestions as to what may be the problem, please let me know!
Cheers,

VCAM mAb troubleshooting
« on: March 30, 2005, 06:39:28 AM »

Offline richard03

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mAb troubleshooting
« Reply #1 on: March 30, 2005, 08:48:06 PM »
These markers usually work better on enzyme digestion pretreatment. You may want to try that such as proteinase K digestion, etc.

Richard

mAb troubleshooting
« Reply #1 on: March 30, 2005, 08:48:06 PM »