1.Preparation of the human blood samples.
For the best results, use whole blood treated with an anticoagulant
such as EDTA, heparin or citrate. We recommend EDTA (K salt) for
the best result. The blood should be used within two hours of
drawing from the donor. The blood samples and the
Polymorphprep™ solution should be at a temperature of 18-22 °C
and during centrifugation also kept within these limits.
2. Preparation of gradient
Layer 5.0 ml of anti-coagulated whole blood over 5.0 ml of
Polymorphprep™ in a 15 ml centrifuge tube or 18 ml of blood over
18 ml Polymorphprep™ in a 50 ml tube. Take care to avoid mixing
of the blood with the separation fluid. Always use equal volumes of
blood and PolymorphprepTM and select a tube that maintains a
similar geometry of layers to the recommended 5 ml + 5 ml in a 15
ml tube.
3. Separation procedure
Centrifuge the tubes at 500-550 gav for 30 minutes in a swing-out
rotor at 18-22 °C. Allow the rotor to decelerate without the break.
Centrifugation for longer times or higher centrifugal force will result
in the PMNs migrating further down towards the pelleted
erythrocytes.
4. Unloading the gradient
After centrifugation, two leukocyte bands should be visible. The top
band at the sample/medium interface will consist of mononuclear
cells and the lower band of PMNs; the erythrocytes are pelleted.
The cell bands are best harvested using a 2 ml syringe attached to
a flat-tipped metal filling cannula (0.8 mm inner diameter). The
PMNs should be diluted by addition of one volume of 0.45% NaCl
solution or culture medium at 0.5 normal concentration in order to
restore normal osmolality. For further dilutions use normal saline or
culture medium.
5. Washing
The suspension is transferred to a 15 ml tube and the cells pelleted
at approx. 400 g for 10 min (18-22 °C). They are resuspended in
saline or culture medium, sedimented again and then resuspended
in a medium compatible with the subsequent analysis.
Description: A sandwich ELISA for quantitative measurement of Human Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Human Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Human Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Rat Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Rat Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Rat Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Canine Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Canine Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Canine Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Porcine Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Porcine Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Porcine Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Goat Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Goat Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Goat Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Mouse Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Mouse Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Mouse Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Rabbit Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Rabbit Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Rabbit Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Monkey Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Monkey Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Monkey Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Pyrex Pear Shaped Separating Funnel with PTFE Stopcock 250ml
Description: A sandwich ELISA for quantitative measurement of Guinea pig Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Guinea pig Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A sandwich ELISA for quantitative measurement of Guinea pig Polymorphonuclear Elastase in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
