Cell Separation and Centrifugation Media

Cell Separation and Centrifugation Media

Products for enhanced cell separation

The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.

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Upon centrifugation, erythrocytes and granulocytes descend through the frit to pellet below the Histopaque®-1077 media. The erythrocytes aggregate, increasing their sedimentation rate, resulting in pelleting at the bottom of the ACCUSPIN™ tube. Lymphocytes and other mononuclear cells, i.e., monocytes, remain above the frit at the plasma- Histopaque®-1077 interface.

The ACCUSPIN™-Histopaque®-1077 System We offer a complete line of products for the separation or extraction of leukocytes, viruses, DNA, RNA, organelles, as well as many other applications. Featured in this product line is the ACCUSPIN™ system; radiation-sterilized centrifuge tubes specially designed with a porous frit to create 2 chambers. Whole blood can be added directly to the ACCUSPIN™ tube without the risk of mixing with the Histopaque®-1077 density gradient media contained in the lower chamber.

ACCUSPIN™ System Tubes Cel

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Product Number

Package Size Name Description

A0561 100 x 15 ml ACCUSPIN™ system tubes, with Histopaque®-1077, sterile (50 ml capacity)

γ-irradiated centrifuge tube, prefilled with Histopaque®-1077 with a porous barrier which separates tube into 2 portions. Blood can be layered into upper portion of the tube and no mixing of blood and the density gradient occurs until the tubes are centrifuged. Suitable for separation of mononuclear cells.

X

A7054 12x 15 ml ACCUSPIN™ system tubes, with Histopaque®-1077, sterile (50 ml capacity)

γ-irradiated centrifuge tube, prefilled with Histopaque®-1077, with a porous barrier which separates tube into 2 portions. Blood can be layered into upper portion of the tube and no mixing of blood and the density gradient occurs until the tubes are centrifuged. Suitable for separation of mononuclear cells.

X

A6929 40 x 3 ml ACCUSPIN™ system tubes, with Histopaque®-1077, sterile (12 ml capacity)

γ-irradiated centrifuge tube, prefilled with Histopaque®-1077, with a porous barrier which separates tube into 2 portions. Blood can be layered into upper portion of the tube and no mixing of blood and the density gradient occurs until the tubes are centrifuged. Suitable for separation of mononuclear cells.

X

A2055 10 each ACCUSPIN™ system tubes, 50 ml capacity, sterile. Holds up to 15 ml of density gradient media.

Empty γ-irradiated centrifuge tube with a porous barrier which separates the tube into 2 chambers, one for the density gradient, the second for the sample. Histopaque® media is added to the tube, which is briefly centrifuged to place it below the frit. Blood can be added to the upper portion of the tube without mixing with the Histopaque® media before centrifugation. Suitable for separation of mononuclear cells.

X

A1805 20 each ACCUSPIN™ system tubes, 12 ml capacity, sterile. Holds up to 3 ml of density gradient media.

Empty γ-irradiated centrifuge tube with a porous barrier which separates the tube into 2 chambers,one for the density gradient, the second for the sample. Histopaque® media is added to the tube, which is briefly centrifuged to place it below the frit. Blood can be added to the upper portion of the tube without mixing with the Histopaque® media before centrifugation. Suitable for separation of mononuclear cells.

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Blood

“Frit”

Histopaque®-1077

Plasma

Centrifuge

10 Minutes

1000 x g

Mononuclear Cells

“Frit”

Red Cells and granulocytes

Blood

Histopaque®-1077

Histopaque®-1119

Plasma

Mononuclear Cells Cells/Platelets (Layer A)

Histopaque®-1077

Granulocytes (Layer B)

Histopaque®-1119

Red Cells

Centrifuge

30 Minutes

700 x g

Blood

Histopaque®-1077

Centrifuge

30 Minutes

400 x gPlasma

Mononuclear Cells

Histopaque®-1077

Red Cells

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Histopaque® density gradient media

Histopaque®-1077: Method: Density Gradient Centrifugation using Sodium Diatrizoate-Polysucrose

An aliquot of Histopaque®-1077 media is carefully layered with blood. The tube is then centrifuged (400 x g) 30 minutes at ambient temperature. A visible layer of mononuclear cells then forms at the Histopaque®-1077/plasma interface. Solution above the mononuclear layer is carefully aspirated and discarded. The mononuclear layer is then recovered, washed and is ready for studies.

Histopaque®-1119: Double Gradient Technique: Sodium Diatrizoate-Polysucrose Gradient 1077 is layered over the 1010 Gradient

Histopaque®-1119 gradient media is used in conjunction with Histopaque®-1077 media, according to a double gradient technique. In this way a layer of cells of the granulocytic series is separated from a zone containing lymphocytes, other mononuclear cells and platelets. To do this, Histopaque®-1077 solution is layered on Histopaque®-1119 solution followed by a layer of blood. After centrifuging (700 x g) for 30 minutes at ambient temperature, two distinct layers of cells become visibly evident. These separate fractions are recovered by aspiration, washed and ready for granulocyte/mononuclear cell studies.

Product Number Package Size Description

10771 100 ml 6 x 100 ml

Histopaque®-1077 - A solution containing polysucrose and sodium diatrizoate, adjusted to a density of 1.077. This medium facilitates the recovery of large numbers of viable mononuclear cells. Centrifugation causes the mononuclear cells to form a distinct layer at the plasma/Histopaque® interface. Aseptically filtered solution. Density: 1.077 g/ml

10831 100 ml 6 X 100 ml

Histopaque®-1083 - A solution similar to Histopaque®-1077, adjusted to a density of 1.083. It permits the separation of mice, rat and other mammalian mononuclear cells. Aseptically filtered solution. Density: 1.083 g/ml

11191 100 ml 6 X 100 ml

Histopaque®-1119 - A solution similar to Histopaque®-1077, adjusted to a density of 1.119. Combined with Histopaque®-1077, it permits the separation of mononuclear cells and neutrophils. Aseptically filtered solution. Density: 1.119 g/ml

• Optimal recoveries 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 and endotoxin-tested filtered solutions • Stable at 2-8 °C for at least 2 years when

protected from light

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Cell Separation (Centrifugation) Media Reagents Cel

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Product Number Package Size Name Description

C301125 g, 50 g, 100 g, 250 g, 500 g, 1 kg

Cesium Chloride, grade I, 99.0%

May be used for the separation of RNA, DNA, ribosomal subunits, proteins, glycoproteins and viruses. ✔ ✔

C6914 500 g, 1 kg Cesium Chloride, grade II, 98.0%

C3139 25 g, 100 g, 500 g, 1 kg

Cesium Chloride, optical grade, ≥99.5%

C3309 10 g, 50 g, 250 g

Cesium Chloride, SigmaUltra, >99.59'0

C2139 100 mg, 500 mg, 1 g, 5 g Collagenase Enzyme used to release adipocytes & hepatocytes from tissue. ✔

C6885 25 mg, 100 mg, 500 mg, 1 g, 5 g Collagenase Enzyme used to release physiologically acitve epididymal

adipocytes from rat tissue. ✔

C5138 25 mg, 100 mg 500 mg, 1 g, 5 g Collagenase Enzyme used to release physiologically active hepatocytes

from rat tissue. ✔

C9263 25 mg, 100 mg 500 mg, 1 g, 5 g Collagenase Enzyme used to release physiologically active pancreatic

islets from rat tissue. ✔

C7657 25 mg, 100 mg, 500 mg, 1 g, 5 g Collagenase Enzyme used to release physiologically active pancreatic

islets from rat tissue. ✔

C6079 500 mg, 5 g Collagenase with Protease Inhibitor

Enzyme and protease inhibitor specially formulated for release of rat pancreatic islet cells from tissue. ✔

D8802 25 ml, 50 ml Dextran 500, 20% solution Used to prepare leukocyte rich plasma. ✔

D6924 1 g, 10 g, 50 g Dextran Sulfate, MW 9,000-20,000

Routinely used to selectively precipitate lipoproteins. ✔

D4911 1 g, 10 g, 50 g, 100 g

Dextran Sulfate, MW 6,500-10,000

D9268 50 g Diatrizoic AcidDiatrizoic acid can be used with PolySucrose 400 to produce density gradients capable of purifying living cells and cell fragments.

✔

F4375 10 g, 25 g, 100 g, 500 g Ficoll® PM 400

Can be used to increase the sedimentation rate of red blood cells. Acts as a stabilizing agent in protein solutions. Used to purify living cells and cell fragments. Useful in preparing various density gradients and high density solutions.

✔ ✔

F9378 5 g, 10 g, 25 g, 100 g, 500 g Ficoll® 400 DL lyophilized powder

F8016 5 g, 100 g, 500 g

Ficoll® 400 DL lyophilized powder, cell culture suitable for cell culture

F1418 25 g, 100 g Ficoll® 400 DL lyophilized powder, SigmaUltra

F2637 5 g, 10 g, 25 g, 100 g

Ficoll® 400 for molecular biology, lyophilized powder

F8636 25 gFicoll® 400 lyophilized powder, Al-irradiated Suitable for cell culture

F5415 25 ml, 50 ml Ficoll® type 400 20% solution

F2878 5 g, 25 g, 100 g, 500 g Ficoll® 70

D2158 100 g Histodenz™ (Nycodenz)

Non-ionic autoclavable, universal centrifugation medium suitable for the isolation of macromolecules, nucleoproteins, cell organelles, and a wide variety of cell types and viruses.

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10771 100 ml, 500 ml, 6 x 100 ml

Histopaque®-1077, density 1.077 g/ml, radiation sterilized

Solution of polysucrose and sodium diatrizoate used to separate mononuclear cells from human peripheral blood or bone marrow, isolation of lymphocytes, and isolation of breast cancer circulating tumor cells from whole blood.

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Cell Separation (Centrifugation) Media Reagents Cel

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Product Number Package Size Name Description

H8889 100 ml, 500 mlHistopaque®-1077, Hybri-Max™ liquid, sterile-filtered, suitable for hybridoma

Solution of polysucrose and sodium diatrizoate- This medium facilitates rapid recovery of viable lymphocytes from small volumes of whole blood. This technique is suitable for use in cell mediated lympholysis and for human lymphocyte antigen (HLA)2 typing. It may also be employed as the initial isolation step prior to enumeration of T-, B-, and ‘null’ lymphocytes.

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10831 100 ml, 6 x 100 ml

Histopaque®- 1083, density 1.083 g/ml, aseptically filled

Solution of polysucrose and sodium diatrizoate used to separate mononuclear cells from rat, mouse or other mammalian peripheral blood or bone marrow.

✔

11191 100 ml, 6 x 100 ml

Histopaque®-1119, density 1.119 g/ml, aseptically filled

Solution of polysucrose and sodium diatrizoate used separate mononuclear cells from peripheral blood or bone marrow. May be able to separate live from dead cells in cell culture.

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H6382 250 g, 1 kg Hydroxyethyl Starch Prepared from corn starch. Sedimenting reagent used in the separation of neutrophils fröm red blood cells. ✔

213233 100 g, 500 g, 1 kg, 12 kg Lithium Chloride ReagentPlus®, 99% May be used to selectively

precipitate RNA ✔

L0650 100 g, 250 g, 500 g

Larcoll™ (highly purified larch arabinogalactan)

From larch wood. Used to prepared density gradients suitable for separation of red blood cells, viruses and organelles.

✔ ✔ ✔

M5266 10 g, 25 g, 100 g Meglumine Diatrizoate

Meglumine diatrizoate is used in conjunction with sodium diatrizoate to separate a variety of cell types and bacterial spores.

✔

D1556 250 ml OptiPrep Density Gradient Medium

Sterile, endotoxin tested solution of 60% lodixanol in water. Suitable to separate a wide variety of cell types, viruses, proteins and organelles.

✔ ✔ ✔ ✔

P1644 25 ml, 100 ml, 500 ml, 1 L Percoll® pH 8.9

Non-toxic and aseptically filtered density gradient which may be resterilized. May be used for the separation of cells, subcellular particles and larger viruses. Coated silica product that is almost chemically inert and does not adhere to membranes. Density 1.13 g/ml.

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P4937 25 ml, 100 ml, 500 ml, 1 L

Percoll® pH 8.9 Cell culture tested

Non-toxic and aseptically filled density gradient which may be resterilized.May be used for the separation of cells, subcellular particles and larger viruses. Coated silica product that is almost chemically inert and does not adhere to membranes. Density 1.13 g/ml.

✔ ✔ ✔ ✔ ✔

P7798 100 g, 500 g Polysucrose 400

Nonionic synthetic polymer of sucrose. Can be used to increase the sedimentation rate of red blood cells. Acts as a stabilizing agent in protein solutions. Used to purify living cells and cell fragments. Useful in preparing various density gradients and high density solutions.

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S450610 g, 50 g, 100 g 250 g, 500 g

Sodium Diatrizoate dihydrate

Sodium Diatrizoate is commonly combined with Polysucrose Type 400 to prepare a density gradient with a density of 1.077 g/ml. The 1.077 g/ml density gradient is used for the preparation of mononuclear cells. Sodium diatrizoate may also be used to obtain complete leukocyte suspensions and to isolate and prepare pure thrombocytes and reticulocytes.

✔

S1888 500 g, 1 kg, 5 kg

Sucrose, cell culture tested/ 500 insect cell culture tested

Sucrose possesses a number of ideal gradient properties. Preformed gradients are used for various rate-zonal and isopycnic procedures. May be used for the purification of proteins, nucleic acids, ribosomes, polysomes and even cells if viability is not essential.

✔

S0389 500 g, 1 kg, 5 kg Sucrose, for molecular biology

S5390 1 kg, 5 kg, 10 kg

Sucrose, Grade I plant cell culture tested

S5391 1 kg, 5 kg, 25 kg

Sucrose, Grade II insect cell culture tested

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Cell Separation (Centrifugation) Companion Products Cel

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Product Number

Package Size Name Description

Z358169 1 each Carousel centrifuge tube rack Acrylic with a polystyrene turntable. 17 position for 50 ml centrifuge tubes. A 2 inch hole in the center accommodates pens. ✔

Z358150 1 each Carousel centrifuge tube rack Acrylic with a polystyrene turntable. 27 position for 15 ml centrifuge tubes. A 3-¾ inch hole in the center will hold a 600 ml beaker.

✔

R4261 1 each Centrifuge tube rack Acrylic with non skid feet. 15 position for 15 ml centrifuge tubes. ✔

R4386 1 each Centrifuge tube rack Acrylic with non skid feet. 8 position for 50 ml centrifuge tubes. ✔

R8509 1 each Centrifuge tube rack Epoxy coated steel, 8 position for 50 ml tubes. ✔



F9755 100 ml, 250 ml Fluorinert® FC-40

Liquid mixture of completely fluorinated aliphatic compounds. For possible use as a high density chase solution in density gradient studies. Density is 1.85 g/ml.

✔

F9880 25 ml, 100 ml Fluorinert® FC-70

Liquid mixture of completely fluorinated aliphatic compounds. For possible use as a high density chase solution in density gradient studies. Density is 1.93 g/ml.

✔

Z340391 1 each Gradient Mixer - 15 ml Acrylic, 2 chamber block suitable for use to prepare centrifugation fradients, using sucrose or cesium chloride solutions. ✔




R650410 X 1 L, 2 X 5 L, 10 L, 50 L

RPMI - 1640 Medium Contains L-glutamine. Does not contain sodium bicarbonate. Powdered medium.

Contains L-glutamine. Does not contain sodium bicarbonate. Powdered medium. ✔

WS16 500 ml Wright Stain, Modified (Manual) Wright stain.

Wright stain, 0.3%, buffered at pH 6.8 in methanol, with stabilizers. ✔

WS32 1 L Wright Stain, Modified (Manual) Wright stain.

WS80 2.5 L Wright Stain, Modified (Manual) Wright stain.

WS128 4 L Wright Stain, Modified (Manual) Wright stain.

MilliporeSigma 400 Summit DriveBurlington, MA 01803

SigmaAldrich.com/histopaque

To place an order or receive technical assistance in the U.S. and Canada, call toll-free 1-800-645-5476 For other countries across Europe and the world, please visit: SigmaAldrich.com/offices For Technical Service, please visit: SigmaAldrich.com/techservice Safety-related Information: SigmaAldrich.com/safetycenter

© 2018 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved. MilliporeSigma, the vibrant M, Sigma-Aldrich, ACCUSPIN, Histopaque, Histodenz, Hybri-Max, and ReagentPlus are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources.

Lit. No. MS_BR2681EN Ver. 0.12018-1361910/2018

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Cell Separation and Centrifugation Media