Product: APC Anti-human CD41a
Cat. Ref: 71-HCD41-A
Reagent provided: 100 test
Description: Monoclonal Mouse Anti-Human CD41a APC is recommended for use in flow cytometry for
identification of Erythroblast. The conjugate is provided in liquid form in buffer containing Antibody Stabilizer
Solution, PBS 20 mM, 0,1% NaN3 and pH 7,2
Clone: HIP8
Isotype: IgG1
HLDA: 4th International Workshops on Human Leucocyte Differentiation, WS Code 38
Fluorochrome: Allophycocyanin (APC)
Reactivity: The monoclonal antibody recognizes the intact platelet gpIIb/IIIa complex, i.e. it does not bind to
dissociated gpIIb or gpIIIa. The antibody does not bind to platelets from patients with Glanzmann Thrombasthenia
and does not recognize the vitronectin receptor, which contains gpIIa as a beta subunit (cf. CD61). The monoclonal
antibody activates human thrombocytes and induces thrombocyte aggregation in the absence of thrombocyte
inhibitors such as EDTA.
Specificity: This clone has been clustered to CD41 in the fourth international Workshop on Human White Cell
differentiation Antigens. The antibody recognises the intact platelet gpIIb/IIIa complex; it does not bind to
dissociated gpIIb or gpIIIa only to gpIIb in the intact complex. This is found on platelets, megakaryocytes and
monocytes. Acute megakaryoblastic l leukaemia cells are positive for the gpIIb/IIIa antigen whereas common
myeloid leukaemias are negative. CD41 is expressed on platelets and megakaryocytes. CD41 is the integrin aIIb
subunit that is expressed as a heterodimer non-covalently associated with the integrin b3 subunit CD61. The
association between CD41 and CD61 is calcium dependent.
Storage: Store in the dark at 2-8 °C. Do not use after expiration date stamped on vial.
Application: It is recommended for use in flow cytometry. This reagent is effective for direct immunofluorescence
staining of human tissue for flow cytometric analysis using 20 μl/106 cells.
Precautions:
1. The device is not intended for clinical use including diagnosis, prognosis, and monitoring of a disease state, and it
must not be used in conjunction with patient records or treatment.
2. This product contains sodium azide (NaN3), a chemical highly toxic in pure form. At product concentrations,
though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive
build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in
plumbing.
Preparation:
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Transfer 5 µL of anticoagulated (EDTA) blood to a 12 x 75 mm polystyrene test tube (106 cells).
Add 95 µL of PBS (It better that it containing 2% bovine serum albumin) without Fixative mix gently with
a vortex mixer.
Add 20 µL of CD41 APC and mix gently with a vortex mixer. The 20 µL is a guideline only; the optimal
volume should be determined by the individual laboratory.
The recommended negative control is a non-reactive APC-conjugated antibody of the same isotype.
Incubate in the dark at 4 °C for 30 minutes or at room temperature (20-25 °C) for 15 minutes.
Add 0, 2 mL 0.01 mol/L PBS (It betters that it containing 2% bovine serum albumin) and resuspend the
cells by using a vortex mixer.
Analyse on a flow cytometer, and the readings of the cytometer are recorded, bearing in the mind that the
FSC and SSC light dispersion parameters must be on a logarithmic scale.
Revision Nº2
Emission date: 12/02/2013
The histogram below is
biparametric representations
(Side Scatter versus
Fluorescence Intensity) of a
lysate normal whole blood
sample gated on leucocytes.
Staining is with CD41 APC
monoclonal antibody (41A100T). Isotypic control
labeling is not shown.
Cells were analyzed on a BD FACSAria™ II Flow Cytometer (Becton Dickinson, San Jose, CA), using Cell Quest
acquisition software.
References:
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P. Menéndez, M.C. de Cañizo, A. Orfao. Immunophenotypic characteristics of PB-mobilised CD34 +
hematopoietic progenitor cells. J Biol Regul Homeost Agents 2001; 15: 53-61.
Nieswandt,B.,B.Echtenacher,F.P.Wachs,J.Schröder,J.E.Gessner,R.E.Schmidt,G.E.Grau,and
D.N.Mäennel.1999.Acute systemic reaction and lung alterations induced by an antiplatelet integrin
gpIIb/IIIa antibody in mice.Blood 94:684 – 693.
Phillips,D.R.,I.F.Charo,and R.M.Scarborough.1991.GPIIb-IIIa:The responsive integrin.Cell 65:359 -362.
Ferkowicz,M.J.,M.Starr,X.Xie,W.Li,S.A.Johnson,W.C.Shelley,P.R.Morrison,and M.C.Yoder.2003.CD41
expression defines the onset of primitive and definitive hematopoiesis in the murine embryo.Development
130:4393 -4403.
Kieffer,N.,and
D.R.Phillips.1990.Platelet
membrane
glycoproteins:functions
in
cellular
interactions.Annu.Rev.Cell Biol.6:329 – 357.
Bakewell,S.J.,P.Nestor,S.Prasad,M.H.Tomasson,N.Dowland,M.Mehrotra,R.Scarborough,J.Kanter,K.Abe,D
.Phillips,and K.N.Weilbaecher.2003.Platelet and osteoclast beta3 integrins are critical for bone
metastasis.Proc.Natl Acad.Sci.U.S.A.100:14205 -14210
*Note: For research use only. Not for diagnostic use.
Revision Nº2
Emission date: 12/02/2013