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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Informazioni su questo articolo
NACRES:
NA.75
UNSPSC Code:
12352207
Prezzi e disponibilità al momento non sono disponibili
Servizio Tecnico
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Permettici di aiutartisterility
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
L'Histopaque-1077 è un mezzo sterile, testato per l′assenza di endotossine e pronto all′uso. Si tratta di una soluzione contenente polisaccarosio e sodio diatrizoato portata a una densità relativa di 1,077g/mL. Questo mezzo di separazione pronto all′uso facilita un recupero rapido di linfociti e altre cellule mononucleate vitali da volumi piccoli di sangue intero o di midollo osseo.
Application
Histopaque-1077 è stato impiegato per:
- la tipizzazione dell'antigene linfocitario umano (HLA)
- lo stadio iniziale di isolamento prima della conta dei linfociti T, B e ‘null’.
- la preparazione di sospensioni di linfociti puri per i saggi di colture cellulari e citotossicità
- l'isolamento e la purificazione delle cellule mononucleate di sangue periferico (PBMC) e delle cellule mononucleate di midollo osseo (BMMC)
- l'isolamento di linfociti umani
- l'isolamento di cellule tumorali in circolo (CTC) di carcinoma mammario da sangue intero
- Histopaque-1077 viene usato insieme a Histopaque-1119 poiché facilita un rapido recupero di cellule mononucleate e granulociti vitali da piccoli volumi di sangue intero
Biochem/physiol Actions
Il sangue anticoagulato viene stratificato sul mezzo Histopaque-1077. Durante la centrifugazione, il polisaccarosio causa l'aggregazione degli eritrociti, che di conseguenza sedimentano rapidamente. I granulociti diventano leggermente ipertonici, aumentando il loro tasso di sedimentazione, con conseguente accumulo sotto forma di pellet sul fondo della provetta. I linfociti e le altre cellule mononucleate rimangono a livello dell'interfaccia plasma/Histopaque-1077.
Features and Benefits
I numerosi vantaggi di Histopaque®-1077 sono:
- permette di recuperare in maniera ottimale le cellule vitali
- consente la separazione selettiva dei diversi tipi di cellule ematiche
- previene la distorsione delle cellule
- preserva la vitalità delle cellule
- l'interferenza da parte di cellule estranee è minima
- offre prestazione riproducibili tra i lotti
- è filtrato sterilmente e testato per l'assenza di endotossine
- è stabile a 2-8°C per almeno 2 anni se mantenuto al riparo dalla luce
Legal Information
Histopaque is a registered trademark of Merck KGaA, Darmstadt, Germany
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Questo articolo | |||
|---|---|---|---|
| description sterile-filtered, density: 1.077 | description sterile-filtered, density: 1.083 | description liquid, sterile-filtered, BioReagent, suitable for hybridoma | description sterile-filtered, density: 1.119 |
| density 1.077 | density 1.083 | density 1.077 at 25 °C (relative density)(lit.) | density 1.119 |
| sterility sterile-filtered | sterility sterile-filtered | sterility sterile-filtered | sterility sterile-filtered |
| form liquid | form liquid | form liquid | form liquid |
| 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 |
| 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 |
Parola di avvertenza
Danger
Codici di pericolo
Avvertenze di sicurezza
Hazard Classifications
Resp. Sens. 1 - Skin Sens. 1
Classe di stoccaggio
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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