how can we separate plasma and platelates from blood
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by fusion....of cell membrane by certain chemicals..
..process named as :-Plateletpheresis
..process named as :-Plateletpheresis
titli4:
But I need the name of process
generally it is called stem cell technology
Plateletpheresis
this is the process...
perfect
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This protocol describes the isolation of human platelets from whole blood.
To prevent the activation of platelets during the procedure, strong mechanical forces (e.g. fast pipetting,
vigorous shaking) should be avoided. In addition, the platelet inhibitors indicated in the protocol can be used,
but other inhibitors exist as well. It is the researcher’s choice to select inhibitors that are most suitable for their
studies and experimental goals.
Examples of applications for isolated platelets are:
Western blotting, flow cytometry, ELISA and immunocytochemistry/immunofluorescence to investigate cellular
and signaling processes within the platelets. Platelet aggregation and adhesion upon stimulation with various
agents can be studied as well as the release of granule components upon activation.
Reagents
Unless platelets are used in tissue culture experiments, the reagents do not need to be sterile. Store and use all
buffers at 4°C, unless indicated otherwise.
ACD buffer (acid-citrate-dextrose)
39 mM citric acid, 75 mM sodium citrate, 135 mM dextrose, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
CPD buffer (citrate-phosphate-dextrose)
16 mM citric acid, 90 mM sodium citrate, 16 mM NaH2PO4, 142 mM dextrose, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
Platelet wash buffer
10 mM sodium citrate, 150 mM NaCl, 1 mM EDTA, 1% (w/v) dextrose, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
HEP buffer (HEP refers to HEPES in the buffer)
140 mM NaCl, 2.7 mM KCl, 3.8 mM HEPES, 5 mM EGTA, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
Tyrode’s buffer
134 mM NaCl, 12 mM NaHCO3, 2.9 mM KCl, 0.34 mM Na2HPO4, 1 mM MgCl2, 10 mM HEPES, pH 7.4.
Depending on experiment, warm buffer up to room temperature or keep on ice before use.
2X platelet lysis buffer
2% NP40, 30 mM HEPES, 150 mM NaCl, 2 mM EDTA, pH 7.4.
Blood collection notes
Many drugs are known to interfere with platelet studies and platelet function. It is important to ensure
that potential blood donors have not taken such medication (e.g. anti-histamines, aspirin, non-steroidal
anti-inflammatory medication) two weeks prior to the blood draw.
Platelets are recommended to be handled and stored in polypropylene, polyethylene or polycarbonate
#HOPE IT HELPS
To prevent the activation of platelets during the procedure, strong mechanical forces (e.g. fast pipetting,
vigorous shaking) should be avoided. In addition, the platelet inhibitors indicated in the protocol can be used,
but other inhibitors exist as well. It is the researcher’s choice to select inhibitors that are most suitable for their
studies and experimental goals.
Examples of applications for isolated platelets are:
Western blotting, flow cytometry, ELISA and immunocytochemistry/immunofluorescence to investigate cellular
and signaling processes within the platelets. Platelet aggregation and adhesion upon stimulation with various
agents can be studied as well as the release of granule components upon activation.
Reagents
Unless platelets are used in tissue culture experiments, the reagents do not need to be sterile. Store and use all
buffers at 4°C, unless indicated otherwise.
ACD buffer (acid-citrate-dextrose)
39 mM citric acid, 75 mM sodium citrate, 135 mM dextrose, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
CPD buffer (citrate-phosphate-dextrose)
16 mM citric acid, 90 mM sodium citrate, 16 mM NaH2PO4, 142 mM dextrose, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
Platelet wash buffer
10 mM sodium citrate, 150 mM NaCl, 1 mM EDTA, 1% (w/v) dextrose, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
HEP buffer (HEP refers to HEPES in the buffer)
140 mM NaCl, 2.7 mM KCl, 3.8 mM HEPES, 5 mM EGTA, pH 7.4.
Before use, warm up to room temperature and adjust pH if necessary.
Tyrode’s buffer
134 mM NaCl, 12 mM NaHCO3, 2.9 mM KCl, 0.34 mM Na2HPO4, 1 mM MgCl2, 10 mM HEPES, pH 7.4.
Depending on experiment, warm buffer up to room temperature or keep on ice before use.
2X platelet lysis buffer
2% NP40, 30 mM HEPES, 150 mM NaCl, 2 mM EDTA, pH 7.4.
Blood collection notes
Many drugs are known to interfere with platelet studies and platelet function. It is important to ensure
that potential blood donors have not taken such medication (e.g. anti-histamines, aspirin, non-steroidal
anti-inflammatory medication) two weeks prior to the blood draw.
Platelets are recommended to be handled and stored in polypropylene, polyethylene or polycarbonate
#HOPE IT HELPS
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