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Category 'Antibodies'
Status: Unanswered
Views: 104

Successful use of CCR4 antibody for human CD3+ lymphocytes

fiona cunningham (Author)

Friday, February 15, 2019 5:13 PM

Dear all,

I wish to ask some advice about the best clone of CCR4 antibody (which company is best) for human CD3+ lymphocytes - I want to measure CCR4 expression on human T cells. I appreciate any advice if you have experience.

thanks alot

F

Kavya A

Saturday, February 16, 2019 3:27 AM

Hello Fiona,

You can try either L291H4 (Biolegend) or REA279 (Miltenyi Biotec). if you are fixing then first stain the cells.

Best,

Kavya

fiona cunningham (Author)

Sunday, February 17, 2019 2:20 PM

Thanks so much Kavya I am wondering which antibody- fluorochrome combination did you find effective please in order to stain for flow cytometry?

Thanks

Fiona 

fiona cunningham (Author)

Sunday, February 17, 2019 2:27 PM

For example is biolegend PE/Cy7 anti-human CD194(CCR4)Antibody useful or do you have a better recommended one ? I am just learning have very limited exorrexper with flow cytometry or any lab work so any advice is extremely appreciated. 

Thanks

F

Emanuela Rasini

Monday, February 18, 2019 7:51 AM

Hello Fiona,

I'm currently using anti-human PE-Cy7 CCR4 from BD. According to my experience it works well in whole blood samples stained and then lysed.

Good luck!

Best,

Emanuela

Kavya A

Monday, February 18, 2019 12:46 PM

Hi Fiona,

I've used BV605 L291H4 from Biolegend and I'm sure BD would equally work well. I've mainly worked on PBMCs. if you are starting out, I suggest you set up a pilot with atleast two different fluorochromes for the markers of more importance/ rare populations/sub-populations and see how the profile looks. I personally would avoid tandem dyes for such markers and make use of tandems in my panel for any parent population. Hope this helps!!

Best,

Kavya

fiona cunningham (Author)

Tuesday, February 19, 2019 2:45 PM

Dear Kavya,

thank you!

may I ask which fluorochrome did you use with REA279 (Miltenyi Biotec) please (anti-human CCR4 antibody)?
thanks

Fiona.