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: Proteins separation resin is a hydrophobic interaction chromatography resin that can be used in the monoclonal antibody purification (Particle size: 65 μm)[1].
Description: Our StemTAG 96-Well Stem Cell Colony Formation Assay provides a high-throughput method to quantify ES cells in just 7-10 days, and no manual cell counting is required. Once colonies are formed, they may be analyzed in three different ways: 1. Lyse cells, then quantify in a fluorescence plate reader using dye included in the kit; 2. Lyse cells, then quantify alkaline phosphatase activity using reagents provided; or 3. Recover colonies from matrix for further culture or analysis.
Description: Our StemTAG 96-Well Stem Cell Colony Formation Assay provides a high-throughput method to quantify ES cells in just 7-10 days, and no manual cell counting is required. Once colonies are formed, they may be analyzed in three different ways: 1. Lyse cells, then quantify in a fluorescence plate reader using dye included in the kit; 2. Lyse cells, then quantify alkaline phosphatase activity using reagents provided; or 3. Recover colonies from matrix for further culture or 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: StemTAG PCR Primer Set for Stem Cell Characterization includes 7 primer pairs: Oct-4, NANOG, AFP, Flk-1, and NCAM, plus GAPDH and beta-actin as controls.
Taq DNA Polymerase - 1000 units with separate dNTP Mix
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer
Taq DNA Polymerase - 5,000 units with separate dNTP Mix
Description: Intact Genomics Taq DNA Polymerase is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity (1, 2) and a 5´ flap endonuclease activity (3, 4). This product is supplied with 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM. Ideal for primary extension reaction with DNA fragments having dA overhang on 3’ ends.Product Includes:Taq DNA Polymerase10x PCR Buffer with Mg2+5x Magic Enhancer10mM dNTP Mix
Total Protein - Murine Embryonic Stem Cell Line D3
Description: Cell Biolabs? HIF-1 Cell Based ELISA Kit is an immunoassay developed for rapid detection of HIF-1 Alpha in fixed cells. Cells on a microplate are stimulated for HIF-1 Alpha stabilization, fixed, permeabilized, and then neutralized in the well. HIF-1 Alpha is then detected with an anti-HIF-1 alpha antibody followed by an HRP conjugated secondary antibody. Each kit provides sufficient reagents to perform up to a total of 96 assays and can detect HIF-1 Alpha from human, mouse, or rat.
Human Polymorphonuclear Elastase,PMN Elastase ELISA Kit
