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.
Fortfahren mit
10771
Histopaque®-1077
sterile-filtered, density: 1.077
Histopaque®-1077
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NACRES:
NA.75
UNSPSC Code:
12352207
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Technischer Dienst
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Unterstützung erhaltensterility
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
Histopaque-1077 ist ein steriles, endotoxingeprüftes, gebrauchsfertiges Medium. Die Histopaque-Zusammensetzung umfasst Polysaccharose und Natriumdiatrizoat und ist auf eine relative Dichte von 1,077 g/ml eingestellt. Dieses gebrauchsfertige Medium ermöglicht die schnelle Rückgewinnung von lebensfähigen Lymphozyten und anderen mononukleären Zellen aus kleinen Mengen an Vollblut oder Knochenmark.
Application
Anwendungsgebiete von Histopaque-1077:
- HLA-Typisierung (Humane Lymphozyten-Antigene).
- Initialer Isolierungsschritt vor der Auszählung von T-, B- und ‘Null’-Lymphozyten.
- Herstellung von reinen Lymphozytensuspensionen für Zellkultur- und Zytotoxizitätsassays.
- Isolierung und Aufreinigung von mononukleären Zellen des peripheren Bluts (PBMCs) und mononukleären Knochenmarkzellen (BMMCs)[1]
- Isolierung humaner Lymphozyten
- Isolierung von zirkulierenden Brustkrebs-Tumorzellen (CTCs) aus dem Vollblut[2]
- Histopak-1077 wird in Verbindung mit Histopak-1119 verwendet, da es die schnelle Rückgewinnung von lebensfähigen mononukleären Zellen und Granulozyten aus kleinen Mengen Vollblut ermöglicht.
Biochem/physiol Actions
Antikoaguliertes Blut wird auf Histopak-1077 geschichtet. Beim Zentrifugieren werden Erythrozyten durch Polysaccharose aggregiert und setzen sich schnell ab. Granulozyten werden leicht hypertonisch, wodurch sich ihre Sedimentationsrate erhöht, was zu einer Pelletbildung am Boden des Zentrifugenröhrchens führt. Lymphozyten und andere mononukleäre Zellen verbleiben an der Plasma/Histopaque-1077-Grenzfläche.
Features and Benefits
Histopaque®-1077 bietet zahlreiche Vorteile, z. B.:
- optimale Rückgewinnung von lebensfähigen Zellen
- selektive Trennung der Blutzelltypen
- keine Zellverformungen
- Lebensfähigkeit der Zellen bleibt erhalten
- minimale Interferenz durch Fremdzellen
- reproduzierbare Leistung von Charge zu Charge
- sterilfiltriert und endotoxingeprüft
- vor Licht geschützt bei 2–8 °C mindestens 2 Jahre stabil
Legal Information
Histopaque is a registered trademark of Merck KGaA, Darmstadt, Germany
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Dieser Artikel | |||
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| density 1.077 | density 1.083 | density 1.077 at 25 °C (relative density)(lit.) | density 1.077 g/mL |
| sterility sterile-filtered | sterility sterile-filtered | sterility sterile-filtered | sterility aseptically filled |
| technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | hybridoma: suitable, single cell analysis: suitable | technique(s) cell culture | mammalian: suitable |
| form liquid | form liquid | form liquid | form solution |
| Quality Level 400 | Quality Level 400 | Quality Level 400 | Quality Level 400 |
| shelf life Expiry date on the label. | shelf life Expiry date on the label. | shelf life - | shelf life Expiry date on the label. |
signalword
Danger
hcodes
Hazard Classifications
Resp. Sens. 1 - Skin Sens. 1
Lagerklasse
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?
1 answer-
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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Is Histopaque 10771 the recommended density for isolating or separating PBMCs in llama blood?
1 answer-
Yes, Histopaque 1077 (catalog number 10771) is the recommended density for mammalian cells.
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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?
1 answer-
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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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.
1 answer-
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?
1 answer-
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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Does Histopaque®-1077 work for mouse blood, or is Ficoll®-Paque Premium 1.084 a better choice?
1 answer-
While Histopaque 10771 can be used, it is important to note that quality control testing for Histopaque 10771 does not include testing with mouse blood. The preferred products for isolating mononuclear cells from rat and mouse blood are Ficoll Paque Premium 1084 (GE17-5446-02) or Histopaque 10831. These products are specifically formulated for rats, mice, and other small mammals for the higher percentage of cell recovery.
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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?
1 answer-
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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Can histopaque-1077 be used to isolate live tumor cells in suspension from dead cells/cell debris?
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Histopaque 10771 is primarily intended for isolating mononuclear cells from human blood and bone marrow. It is also utilized in cell culture to eliminate dying cells from hybridomas. However, the effectiveness of Histopaque 10771 or Histopaque 10831 in purifying live cells from deceased liver cells remains untested. Percoll and OptiPrep are common alternative methods.
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