The largest database of trusted experimental protocols

Histopaque media

Manufactured by Merck Group

Histopaque® media is a density gradient solution used for the isolation of mononuclear cells from whole blood. It facilitates the separation of different blood cell types by density differentiation during centrifugation.

Automatically generated - may contain errors

3 protocols using histopaque media

1

Acoustic Purification of Lymphocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 5

Blood plasma comprising lymphocytes and non-target cells was subjected to acoustic energy, generally as described above. Prior to flowing the blood plasma through a microfluidic separation channel, plasma samples were pretreated by diluting with an additive comprising a density gradient medium to provide experimental samples having solution densities ranging between about 1.04 and 1.06 (g/mL) and a control sample having a solution density averaging about 1.08 g/mL. Specifically, the samples were pretreated with Histopaque® media (Sigma-Aldrich, St. Louis, Mo.) to regulate their density.

Surprisingly, the samples having a specific density between about 1.04 and 1.06 g/mL exhibited better purification of lymphocytes. These results were surprising because conventional density medium separation by centrifugation teaches a target density closer to 1.08 g/mL, which is the average density of mononuclear cells. Thus, by regulating the biofluid density to be slightly different than the density of the average target cells (here about 0.02 g/mL to 0.04 g/mL less than the average density of lymphocytes) better purification may be achieved.

+ Open protocol
+ Expand
2

Mouse Pancreatic Islet Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mouse pancreatic islet isolation was performed as previously described (Li et al., 2009 (link)) with some modifications. Mice were euthanized in accordance with ASPA 1986 guidelines. The abdominal internal region was exposed and in situ pancreatic digestion was performed: a clamp was positioned on the top of the duodenal papilla to block the passage from the common bile and pancreatic duct into the intestine. Pancreatic digestion was carried out by injecting collagenase-containing media (Low glucose DMEM (Mediatech), 10mM Hepes buffer (Invitrogen), collagenase 0.8 mg/ml (Roche)) into the ductal trees. Inflated pancreata were then collected and incubated at 37oC for 15 minutes in the collagenase media. Islet purification was performed by sequential filtration through a 250μm filter and density gradient cell separation using a combination of Histopaque media (H-1077 and H-1119, Sigma). Single acinar cells and cell clusters smaller than 40μm were removed and pancreatic islets were manually handpicked. After O/N 37oC incubation in culture media, islets were lysed for RNA extraction.
+ Open protocol
+ Expand
3

Isolation of PBMCs from Whole Blood

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole blood was collected in EDTA tubes 24 h prior to humane euthanasia. Using density gradient centrifugation, peripheral blood mononuclear cells (PBMCs) were harvested as previously described (Hida et al., 2002 (link)). Briefly, whole blood was overlayed on a double gradient of 1,077 and 1,119 histopaque media (Sigma-Aldrich). Then, samples were centrifuged at 700 g for 30 min with the enriched PBMC layer carefully removed, flash frozen in liquid nitrogen and stored at −80°C.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!