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There have been 8 items by inas (Search limited from 03-June 19)


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#33121 IP and western blot

Posted by inas on 15 August 2009 - 08:38 AM in SDS-PAGE and Western Blotting

thank you very much for your reply
actully i couldnt understand the mening of your statment, could you please explane more for me.
regards



#32715 IP and western blot

Posted by inas on 12 August 2009 - 01:03 AM in SDS-PAGE and Western Blotting

[Thank you for your reply
the exposure time is very short
and regarding the concintration of the AB isit for IP or WB???



#32664 IP and western blot

Posted by inas on 11 August 2009 - 01:25 PM in SDS-PAGE and Western Blotting

I normally do IP then western blot but my problem is i got an extra band (my band in 52KDa and the extra band 30 KDa)
i followed thr folowing protocol
1- add 5ul of mouse anti human antibody (IP) incubation over night
2- add 20 ul proten G agarose
3-collect immunoprecipitates then wash pellet 4 times with PBS
4- dilute pellet with 30 ul lysis buffer and add sample buffer , boil , centerfuge to pellet the agarose beads
5- SDS page analysis of the supernatant
6-Transfer to nitrocellulose membrane.
7- blocking
8- incubate membrane with mouse anti human antibody( primary)
9- wash the incubate with rabbit ant mouse antobody HPR (secondary)
10- wash
11- ECL
12- X ray film
please help me by answer me why i got an extra band?????

many thanks



#18027 IgE

Posted by inas on 06 March 2009 - 06:06 PM in Immunology

Hi ,
Please i need to buy human IgE labeled with FITC but i dont know from where i can buy it.
anyone can help me


Regards

Inas



#18026 online database for antibodies

Posted by inas on 06 March 2009 - 06:05 PM in Immunology

Hi ,
Please i need to buy human IgE labeled with FITC but i dont know from where i can buy it.
anyone can help me


Regards

Inas



#17074 Dendritic_ cells

Posted by inas on 26 February 2009 - 02:25 PM in Immunology

Please answer me as soon as possible, I realy need help
Regards :o



#17071 Dendritic_ cells

Posted by inas on 26 February 2009 - 02:13 PM in Immunology

Hi ,
My protocol is attached

Thank you for your reply

Attached Files




#16681 Dendritic_ cells

Posted by inas on 23 February 2009 - 03:52 PM in Immunology

hi I am PhD student (Immunology) i am facing problem in lab when i isolat Monocyte from human blood using CD14 beads and generate them to dendritic cells most the cells die during incubation or lose them during the washing step, i need helo in solving that problem because i dont knoe the cause of that . Please

Regards




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