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Choosing the right fluorochrome


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#1 Salem

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Posted 23 July 2011 - 06:03 AM

Dear All,
Greeting, I am targeting several genes in different pathways, upon targeting I'd like to check the differentiation in patients & two cell lines SKNO-1 and Kasumi-1 (AML- t(8;21)) through the following CD markers; CD14, CD11b, CD34 & CD45.
Could you please help me by choosing the right fluorochrome for every CD, taking into account two things the lower overlapping and a good brightness

* Fluorochrome Brightness Index
http://www.biolegend...tness_Index.pdf
* Fluorescence Spectra Analyzer
http://www.biolegend...spectraanalyzer
* CD14 products
http://www.biolegend...h&criteria=cd14
* CD11b products
http://www.biolegend...&criteria=cd11b
* CD34 products
http://www.biolegend...h&criteria=cd34
* CD45 products
http://www.biolegend...h&criteria=cd45

Our machine is BD FACSCanto II (3 Lasers)
Sorry I am new to flow cytometry and I'd like to make sure.

Thank you very much for your help in advance.
Salem

#2 grayhair

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Posted 25 July 2011 - 11:22 AM

Dear All,
Greeting, I am targeting several genes in different pathways, upon targeting I'd like to check the differentiation in patients & two cell lines SKNO-1 and Kasumi-1 (AML- t(8;21)) through the following CD markers; CD14, CD11b, CD34 & CD45.
Could you please help me by choosing the right fluorochrome for every CD, taking into account two things the lower overlapping and a good brightness

* Fluorochrome Brightness Index
http://www.biolegend...tness_Index.pdf
* Fluorescence Spectra Analyzer
http://www.biolegend...spectraanalyzer
* CD14 products
http://www.biolegend...h&criteria=cd14
* CD11b products
http://www.biolegend...&criteria=cd11b
* CD34 products
http://www.biolegend...h&criteria=cd34
* CD45 products
http://www.biolegend...h&criteria=cd45

Our machine is BD FACSCanto II (3 Lasers)
Sorry I am new to flow cytometry and I'd like to make sure.

Thank you very much for your help in advance.
Salem


Although I'm not familiar with those two cell lines and it sounds like you are not also , based on the fact that CD14 and CD11b usually have a heterogeneous pattern of
staining I would use CD14 in FITC. CD11b in PE, CD45 in PerCP and CD34 in APC. Most AML,s are usually positive for CD34 and usually have a
nice cluster and CD45 is usually at a density dimmer than lymphocytes.

#3 Salem

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Posted 10 August 2011 - 08:15 AM

Thank you




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