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
Histopaque®-1077
sterile-filtered, density: 1.077
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100 ML
$52.83
6 X 100 ML
$192.00
500 ML
$201.00
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100 ML
$52.83
6 X 100 ML
$192.00
500 ML
$201.00
About This Item
UNSPSC Code:
12352207
NACRES:
NA.75
Recommended Products
sterility
sterile-filtered
Quality Level
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 is a sterile, endotoxin tested, ready-to-use medium. The Histopaque composition includes polysucrose and sodium diatrizoate, adjusted to a relative density of 1.077g/mL. This ready-to-use medium facilitates rapid recovery of viable lymphocytes and other mononuclear cells from small volumes of whole blood or bone marrow.
Application
Histopaque-1077 has been used for:
- human lymphocyte antigen (HLA) typing
- initial isolation step prior to the enumeration of T, B, and ‘null’ lymphocytes.
- preparation of pure lymphocyte suspensions for cell culture and cytotoxicity assays.
- isolation and purification of peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMCs)[1]
- isolation of human lymphocytes[2]
- isolation of breast cancer circulating tumor cells (CTCs) from the whole blood[3]
- Histopaque-1077 is used in conjunction with Histopaque-1119 as it facilitates the rapid recovery of viable mononuclear cells and granulocytes from small volumes of whole blood
Features and Benefits
Histopaque®-1077 offers numerous advantages, such as:
- Optimal recovery of viable cells
- Permits selective separation of blood cell types
- Prevents cell distortion
- Preserves cell viability
- Minimal extraneous cell interference
- Reproducible performance from batch to batch
- Sterile-filtered and endotoxin-tested
- Stable at 2-8°C for at least 2 years when protected from light
Principle
Anticoagulated blood is layered onto Histopaque-1077. During centrifugation, erythrocytes are aggregated by polysucrose and rapidly sediment. Granulocytes become slightly hypertonic, which increases their sedimentation rate, resulting in pelleting at the bottom of the centrifuge tube. Lymphocytes and other mononuclear cells remain at the plasma/ Histopaque-1077 interface.
Legal Information
Histopaque is a registered trademark of Merck KGaA, Darmstadt, Germany
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signalword
Danger
hcodes
Hazard Classifications
Resp. Sens. 1 - Skin Sens. 1
Storage Class
12 - Non Combustible Liquids
wgk_germany
WGK 3
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Articles
Centrifugation enables the separation of particles by sedimentation. Learn how to separate particles using a centrifuge and how to use Stokes' law to calculate the velocity of sedimentation.
Protocols
This troubleshooting guide addresses the most common sources of error observed when using Histopaque but is not meant to be a comprehensive list.
Ficoll 400® is a non-ionic polymer with high hydrophilicity and water solubility, suitable for various biological applications.
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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?
1 answer-
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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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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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Can Histopaque 10771 be utilized to separate tumor cells from monocytes?
1 answer-
Histopaque 10771 is unlikely to be suitable for separating tumor cells from monocytes. It is designed to separate monocytes and lymphocytes from human blood. Unless the density of your specific tumor cells significantly differs from 1.077 g/ml, there is a possibility that tumor cells may be collected within the mononuclear cell band. Alternatives such as Percoll, Percoll Plus, or OptiPrep could be better options.
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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?
1 answer-
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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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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What is the percentage of peripheral blood mononuclear cells (PBMCs) recovered from human blood?
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Generally, Histopaque recoveries typically range from 50% to 80%. The percentage of recovery varies significantly depending on the total white cell count of the patient. Higher white cell counts typically have the best recovery rates. Yields are also generally higher with fresh blood and lower with older blood.
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Can histopaque-1077 be used to isolate live tumor cells in suspension from dead cells/cell debris?
1 answer-
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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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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