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isolation of b cells from PBMNCs

isolation of b cells from PBMNCs - Flow Cytometry Forum

isolation of b cells from PBMNCs - Flow Cytometry Forum. Discuss FACS cell sorting, flow cytometry, analysis and cell staining.


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Old 01-24-2012, 09:23 AM
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Question isolation of b cells from PBMNCs



Hi everyone, i have to isolate b cells with facs from PBMCs and i am not sure which antibodies i should use to really get out all b cells (also plasmacells, plasmablast, mature...)

I know that i have to use Cd 45 for WBCs and CD 19 as a general b cell marker but i think i wont get out the later b cells like plasmacells. I use FACS ARIA 3.
Any ideas or experiences?

greetings
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Old 01-24-2012, 08:18 PM
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Default Re: isolation of b cells from PBMNCs

Quote:
Originally Posted by Vanweinberg View Post
Hi everyone, i have to isolate b cells with facs from PBMCs and i am not sure which antibodies i should use to really get out all b cells (also plasmacells, plasmablast, mature...)

I know that i have to use Cd 45 for WBCs and CD 19 as a general b cell marker but i think i wont get out the later b cells like plasmacells. I use FACS ARIA 3.
Any ideas or experiences?

greetings
CD45/B220 should get plasma cells. Syndecan-1 is also a plasma cell marker. How come you want everything? Usually people go for a specific subset. You might want to search for memory B cell markers. Also are you going for B1 cells or just B2 ?
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  #3  
Old 01-25-2012, 08:04 AM
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Default Re: isolation of b cells from PBMNCs

I want to do IGH sequencing, so in fact it would be ok to use PBMCs without additional b cell sorting because the primer for pcr will hopefully bind only to b cells which have a already recombinated heavy chain. Nevertheless my lab chief wants me to do b cell sorting with FACS bevor...So my idea is to get out simply all b cells to make sure that i don't lose any useful cells.
Concerning B1 cells, i think i've read that there are no B1 cells in the peripheral blood?
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Old 02-26-2012, 04:41 PM
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Default Re: isolation of b cells from PBMNCs

There are B1 cells in peripheral blood. The older you are the higher B1 cell count you have.
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Old 03-15-2012, 10:17 PM
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Default Re: isolation of b cells from PBMNCs

I think CD19 is expressed by all B cell stages if am not mistaked.
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Old 07-02-2012, 03:43 PM
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Default Re: isolation of b cells from PBMNCs

Quote:
Originally Posted by habdelsa View Post
I think CD19 is expressed by all B cell stages if am not mistaked.
Yep. From early pro-B cells.
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b cells , b-cells , cells , isolation , pbmc , pbmcs , pbmncs


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