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
Histopaque®-1077
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| Pack Size | SKU | Availability | Price |
|---|---|---|---|
| 100 mL | Check Cart for Availability | $94.40 | |
| 500 mL | Check Cart for Availability | $317.00 | |
| 1 gal | Please contact Customer Service for Availability | $370.00 | |
| 6 x 100 mL | Check Cart for Availability | $380.00 |
About This Item
NACRES:
NA.75
UNSPSC Code:
12352207
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
- isolation of breast cancer circulating tumor cells (CTCs) from the whole blood[2]
- 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
Biochem/physiol Actions
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.
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
Legal Information
Histopaque is a registered trademark of Merck KGaA, Darmstadt, Germany
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This Item | |||
|---|---|---|---|
| form liquid | form liquid | form liquid | form solution |
| 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 |
| 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 |
| 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
Storage Class
12 - Non Combustible Liquids
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Protocols
Ficoll 400® is a non-ionic polymer with high hydrophilicity and water solubility, suitable for various biological applications.
This troubleshooting guide addresses the most common sources of error observed when using Histopaque but is not meant to be a comprehensive list.
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.
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Global Trade Item Number
| SKU | GTIN |
|---|---|
| 10771-10L | 04061837279089 |
| 10771-6X100ML | 04061838260642 |
| 10771-1GA | 04061837279096 |
| 10771-100ML | 04061838260628 |
| 10771-500ML | 04061838260635 |
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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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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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Will Histopaque 10771 be suitable for isolating CD45-positive lymphocytes from mouse whole blood?
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While Histopaque 10771 can be used for isolating mouse mononuclear cells including lymphocytes and monocytes, the formulation may not be optimal. Alternatively, Histopaque 10831 is available, with a density of 1.083 g/ml and Ficoll Paque 1084 with a density of 1.084 g/mL. These formulations are specifically designed for the purification of mononuclear cells in rats, mice, and small mammals.
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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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