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Merck

10771

Histopaque®-1077

sterile-filtered, density: 1.077

Histopaque®-1077

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About This Item

UNSPSC Code:
12352207
NACRES:
NA.75

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sterility

sterile-filtered

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

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This Item
10831H8889A6929
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

form

liquid

form

liquid

form

liquid

form

solution

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.

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

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

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Hazard Classifications

Resp. Sens. 1 - Skin Sens. 1

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3


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Evaluation of respiration with clark type electrode in isolated mitochondria and permeabilized animal cells.
Silva AM and Oliveira PJ
Methods in Molecular Biology, 810, 7-24 (2012)
Romain A Colas et al.
Journal of immunology (Baltimore, Md. : 1950), 197(11), 4444-4452 (2016-11-02)
Maresin 1 (MaR1) is an immunoresolvent that governs resolution of acute inflammation, and its local metabolism in the context of infectious inflammation is of interest. In this study, we investigated the MaR1 metabolome in infectious exudates and its bioactions in
Erin E Thacker et al.
Vaccine, 27(50), 7116-7124 (2009-09-30)
Targeting viral vectors encoding tumor-associated antigens to dendritic cells (DCs) in vivo is likely to enhance the effectiveness of immunotherapeutic cancer vaccines. We have previously shown that genetic modification of adenovirus (Ad) 5 to incorporate CD40 ligand (CD40L) rather than
Maria Björkqvist et al.
The Journal of experimental medicine, 205(8), 1869-1877 (2008-07-16)
Huntington's disease (HD) is an inherited neurodegenerative disorder characterized by both neurological and systemic abnormalities. We examined the peripheral immune system and found widespread evidence of innate immune activation detectable in plasma throughout the course of HD. Interleukin 6 levels
Ana C Leal et al.
Scientific reports, 7(1), 6438-6438 (2017-07-27)
Cancer patients are at an increased risk of developing thromboembolic complications. Several mechanisms have been proposed to explain cancer-associated thrombosis including the release of tumor-derived extracellular vesicles and the activation of host vascular cells. It was proposed that neutrophil extracellular

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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.

Questions

1–10 of 22 Questions  
  1. 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
    1. 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.

      Helpful?

  2. 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
    1. 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.

      Helpful?

  3. Does Histopaque®-1077 work for mouse blood, or is Ficoll®-Paque Premium 1.084 a better choice?

    1 answer
    1. 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.

      Helpful?

  4. Is Histopaque 10771 the recommended density for isolating or separating PBMCs in llama blood?

    1 answer
    1. Yes, Histopaque 1077 (catalog number 10771) is the recommended density for mammalian cells.

      Helpful?

  5. 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
    1. 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.

      Helpful?

  6. What is the percentage of peripheral blood mononuclear cells (PBMCs) recovered from human blood?

    1 answer
    1. 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.

      Helpful?

  7. Can histopaque-1077 be used to isolate live tumor cells in suspension from dead cells/cell debris?

    1 answer
    1. 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.

      Helpful?

  8. 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
    1. 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.

      Helpful?

  9. Can Histopaque 10771 be utilized to separate tumor cells from monocytes?

    1 answer
    1. 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.

      Helpful?

  10. 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
    1. 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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1–10 of 22 Questions  

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