Basophills summary

Efros, Lyubov LEfros at pdl.com
Fri Mar 22 14:41:20 EST 2002


  Hello all,
I want to appreciate everyone who responded to my quire about basophills!
 Here the summary from all your replays.
  Thanks  a lot, Cytometry Mailing List is definitely a great source
  Lyuba
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BeckmanCoulter now has available an antibody for basophils.  It is CD203c
and it comes available in PE.  It is specific for basophils, and is also
fantastic for basophil
activation studies.
Fields of application may include:
    * Identification and differentiation of basophils and mast cells
    * Basophil activation in allergic and inflammatory responses
    * Studies of stem cell differentiation and maturation
    * Lineage assessment in hematopoietic disease

Main Features include:
    * It is a specific marker for the mast cell/basophil lineage
    * Sufficient resolving power to distinguish resting basophils in whole
blood
    * Three to ten fold upregulation after activation of basophils
    * Marks basophil activation as a marker for IgE mediated allergy
    * Sufficient as a stand-alone marker for basophils, there is no need to
combine it with any other marker to identify basophils

If you are interested, your reps name is Pam Silveraj.	She can be reached
at
    1-800-338-8830  EXT: 8887

regards

Dave Osborn    BeckmanCoulter Cytometry Account Manager
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I am working on basophils from peripheral blood. This reference might be of
interest for you
Best wishes
Frequency and characterization of antigen-specific IL-4 and IL-13 producing
basophils and T cells in peripheral blood of healthy and asthmatic subjects
G.Devouassoux, B.Foster, LM.Scott, DD.Metcalfe, C.Prussin.
J Allergy Clin Immunol 1999;104:811-9.

Dr. Gilles Devouassoux
E-Mail GDevouassoux at chu-grenoble .fr
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We identify basophils from their 45/ssc properties (in the lymphoid
population) and immunophenotyping - CD9+33+13+22+19-.
 Not too sure how you might go about enriching them

Beth Rees
Flow Cytometry Lab
Pathology Dept
Royal Hobart Hospital
Hobart, Tasmania
Australia
----------------------------------------------------------------------Depend
s what you want to do. If you want to look at their function I would use the
BASO-test from www.orpegen.de. The kit saves you a lot of hassle with
regards to titration and preparation of standards.
Regards
Gerhard
----------------------------------------------------------------------There
are no basophil specific surface markers. There are 2 basophil granule
specific markers, BB1 and 2D7, which can be used with I/C staining. .
Because of the small numbers of basophils (about 1%) we have generally use 2
markers to identify them. We stain with anti-IgE and a panel of lineage
markers (CD2, 14, 16, 19). IgE+, lin- cells are >99% basophils. MHC II
positive dendritic cell precursors make up the balance.

Calman
<<DEVOUASSOUX1999.JACI>>
----------------------------------------------------------------------You
may find a protocol for staining dendritic cells from human PBMC and whole
blood on Pharmingen's web site at
http://www.bdbiosciences.com/immunocytometry_systems/application_notes/pdf/r
eagapp3.pdf.  The protocol shows staining of basophils vs. CD11c vs.
CD123dendritic cells.
Good luck,
Andy Morschauser
Schering-Plough Research Institute

----------------------------------------------------------------------
depends what you want to do with the basophils. We used FITC labeled IgE and
sorted the basophils on a high speed sorter with good success. FITC-IgE
labeled cells can also be purified using anti-FITC-magnetic beads from
Miltnyi biotech(Auburn, CA).
I hope this helps.
Ruedi

Dr. Ruedi Braun
Flow Facility Manager and Associated Scientist
Ottawa Health Research Institute
501 Smyth Road
Ottawa, K1H 8L6
Canada

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Attached pls find our basophil combination(pdf file), which works really
well in degranulation assays.

Rudi  (See attached file: Baso02-7900030-27pg14-15_r1.pdf)
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I understand that blood basophils express CD123 (IL-3-R). I use the BDIC kit
to label dendritic cell precursors and clearly see this population.

charles
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CD203c is a good anti-basophil surface marker in whole blood.
call BC 800 352 3433

Martin J Dominguez
Cytomics
Beckman Coulter UK
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you can use a fycoll hypaque prep called Mono-poly from ICN which separates
pbmcs, granulocytes and rbcs in one tube
Adrian Rubio [arubio at biotechimaging.com]
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It is tricky, but they can be detected.  Basophils are CD123+, HLADR-.
You will need to eliminate the CD19+, CD3+, CD14+, CD16+ cells out of the
picture and work with the Lin negative or weak positive cell population.
Look in the BD Biosciences web site...www.bdbiosciences.com.  Look for a
technical note for peripheral blood dendritic cells.  In that process, in
their gating strategy, you can look at basophils too.
I hope this helps

Enoc Hollemweguer
Program Manager
Clinical Research
BD Biosciences Pharmingen
San Diego, CA
858-812-8922
----------------------------------------------------------------------
     Dear Flow-ers,
	I am looking  if someone have experience using different cell
surface markers for  basophils from Human peripheral whole blood (without
preparing PBMC). Also,if it possible to do enrichment  of basophils,without
separating mononuclear cells first.
	    Any information is greatly appreciated.

    Lyuba Efros
    Preclinical Department
   PDL, Fremont, CA
    lefros at pdl.com

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