Histopaque 10771 was introduced in the late 1970s or early 1980s. As such, there is a long history of shipping the product when the temperature falls below freezing. It is relatively common for the bottles to freeze during ground shipping during the winter months. The standard advise is to thaw the bottles at either room temperature at a 37°C. Once the bottle has thawed, it will be necessary to invert the bottle multiple times to ensure the gradient is uniform. If the bottle is not inverted multiple times, a gradient in situ will form as freezing is one method of forming gradients. As long as the solution is clear and the bottle has not broken, the product is suitable for use.
10771
Histopaque®-1077
sterile-filtered, density: 1.077
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A propos de cet article
NACRES:
NA.75
UNSPSC Code:
12352207
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Laissez-nous vous aidersterility
sterile-filtered
Quality Segment
form
liquid
shelf life
Expiry date on the label.
technique(s)
cell culture | mammalian: suitable
impurities
endotoxin, tested
density
1.077
application(s)
hematology
histology
storage temp.
2-8°C
General description
Le produit Histopaque-1077 est un milieu stérile prêt à l'emploi dépourvu d'endotoxines (tests à l'appui). Sa composition, de densité relative ajustée à 1,077 g/ml, inclut du polysaccharose et du diatrizoate de sodium. Ce milieu prêt à l'emploi permet de récupérer rapidement les lymphocytes viables et d'autres cellules mononucléées à partir de petits volumes de sang total ou de moelle osseuse.
Application
L'Histopaque-1077 a été utilisé dans les situations suivantes :
- typage d'antigènes de lymphocytes humains (HLA).
- première étape d'isolement avant le dénombrement de lymphocytes T, B et "nuls".
- préparation de suspensions de lymphocytes pures pour la culture cellulaire et des tests de cytotoxicité.
- isolement et purification de cellules mononucléées de sang périphérique (PBMC) et de cellules mononucléées de moelle osseuse (BMMC).
- isolement de lymphocytes humains.
- isolement de cellules tumorales circulantes de cancer du sein (CTC) à partir de sang total.
- La solution Histopaque-1077 est utilisée avec Histopaque-1119 pour récupérer rapidement les cellules mononucléaires et les granulocytes viables présents dans de petits volumes de sang total.
Biochem/physiol Actions
Étaler du sang traité avec un anticoagulant sur Histopaque-1077. Durant la centrifugation, agréger les érythrocytes avec du polysaccharose ; ils sédimentent alors rapidement. Les granulocytes deviennent légèrement hypertoniques, ce qui accroît leur vitesse de sédimentation et entraîne la formation d'un culot au fond du tube à centrifuger. Les lymphocytes et les autres cellules mononucléées restent à l'interface plasma/Histopaque-1077.
Features and Benefits
Histopaque®-1077 présente de nombreux avantages, notamment :
- Recouvrement optimal des cellules viables
- Permet de séparer sélectivement les différents types de cellules sanguines
- Évite la déformation des cellules
- Préserve la viabilité cellulaire
- Interférences minimales avec les cellules exogènes
- Performances reproductibles d'un lot à l'autre
- Produit stérilisé par filtration et trouvé négatif pour la recherche d'endotoxines
- Stable entre 2 et 8 °C à l'abri de la lumière pendant au moins 2 ans.
Legal Information
Histopaque is a registered trademark of Merck KGaA, Darmstadt, Germany
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Cet article | |||
|---|---|---|---|
| description sterile-filtered, density: 1.077 | description sterile-filtered, density: 1.083 | description liquid, sterile-filtered, BioReagent, suitable for hybridoma | description sterile-filtered, density: 1.077 g/mL |
| density 1.077 | density 1.083 | density 1.077 at 25 °C (relative density)(lit.) | density 1.077 g/mL |
| form liquid | form liquid | form liquid | form solution |
| technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) single cell analysis: suitable, cell culture | hybridoma: suitable | technique(s) cell culture | mammalian: suitable |
| sterility sterile-filtered | sterility sterile-filtered | sterility sterile-filtered | sterility aseptically filled |
| Quality Level 400 | Quality Level 400 | Quality Level 400 | Quality Level 400 |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C |
signalword
Danger
hcodes
Hazard Classifications
Resp. Sens. 1 - Skin Sens. 1
Classe de stockage
12 - Non Combustible Liquids
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Hello, we had a problem with the refrigerator temperature dropping below 4 degrees Celsius for over the weekend. Can the Histopaque-1077 stored in this refrigerator still be used?
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Hello, we had a problem with the refrigerator leading the temperature increasing to 44 degrees Celsius for over the 2 days of weekend, so can our Histopaque-1077 stored in this refrigerator can be used anymore?
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This product ships at ambient temperature and is stable for relatively short periods at room temperature. There is a 1 - 2 week allowance for transit time. A storage temperature excursion of 2 days is not expected to impact the quality or performance of this material.
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What impact does freezing Histopaque or Ficoll solutions have on PBMC purification? Will the freeze/thaw cycle affect product performance in assays, or will it continue to function as expected?
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It is recommended to avoid freezing Histopaque or Ficoll Paque solutions. Thawing can be achieved by leaving the bottles at room temperature, followed by gentle inversion to mix the solution after thawing.If the solution is clear, particle-free, and the bottle is intact, the product is suitable for use. However, if the bottle is cracked or broken, it should be replaced.
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Can we use a ratio of blood to separation media of 4:3 with Histopaque-1077? Also, can Histopaque-1077 be used to dilute the blood up to 1:2 before layering with separation media with HBSS buffer, and then wash the cells too.
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Histopaque 10771/H8889 and Ficoll Paque solutions are produced with similar reagents. As Histopaque has a slightly lower density range, yields might be slightly lower. Therefore, it is a mixed bag and yields with Histopaque might be slightly lower. When observing a very high white cell count, dilution is recommended. The standard dilution for peripheral blood is generally 1:2. For bone marrow, a 1:2 or 1:4 dilution are more commonly used. The expectation is that cell recovery should be almost the same.
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Do you have a recommended protocol for purifying PBMCs from larger volumes using Histopaque 1077 in 50 mL tubes instead of 15 mL tubes?
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For PBMCs, the catalog numbers for Histopaque 10771, H8889, and 10831 are applicable. The Instructions For Use indicate that a 15 ml centrifuge tube with 3 ml of Histopaque 10771 is recommended. This procedure can also be used for a 50 ml centrifuge tube, considering that the height of both tubes is comparable while their diameter differs. To accommodate a 50 ml centrifuge tube, it is advised to use 15 ml of Histopaque 10771. No alterations are needed for the centrifugation time and washing steps. It is sufficient to wash the isolated cells with isotonic PBS or cell culture media.
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What is the efficacy and reliability of using Histopaque®-1077 for separating low-density neutrophils (LDN) and normal-density neutrophils (NDN) from high-density neutrophils?
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The Histopaque 1077 procedure is not designed to purify low-density or high-density neutrophils. Instead, Histopaque 10771 is intended for purifying Peripheral Blood Mononuclear Cells (PBMCs). When using Histopaque 10771, it is expected that the collected mononuclear cells would contain 5% or less of neutrophils, likely in the form of low-density neutrophils.
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Are Histopaque 1077 and Ficoll Paque pure products? What is the percentage of polysaccharide or polysucrose in Histopaque 1077 and Ficoll Paque?
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Both Histopaque and Ficoll Paque are formulated with a combination of polysucrose 400 and sodium diatrizoate. On average, Histopaque and Ficoll Paque solutions contain approximately 5.7% polysucrose and 9.0% sodium diatrizoate. The final density of these solutions is expected to be around 1.077 g/ml for both products with a typical density variation of +/- 0.001%.
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If Histopaque-1077 is brought to room temperature, how long can it be left at that temperature before use? Specifically, will leaving it for a few hours cause any issues?
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Ficoll Paque has a storage temperature range of 4°C to 30°C for unopened bottles, while opened bottles should be stored at 4°C to 8°C. However, Histopaque can be stored at room temperature for short periods.
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Can Histopaque 10771 be placed in a 37°C water bath to bring it to room temperature after being taken out of the refrigerator?
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Histopaque 10771 is normally stored at 2-8°C for long-term storage. However, it can be shipped at ambient temperature, suggesting that it is not easily affected by short-term storage conditions. Therefore, warming it up to 37°C is unlikely to have any detrimental effects on the product.
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Is Histopaque 10771 the recommended density for isolating or separating PBMCs in llama blood?
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Yes, Histopaque 1077 (catalog number 10771) is the recommended density for mammalian cells.
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