The general ratio of blood to Histopaque is 1 to 1. For 0.5ml of mouse blood, it is recommended to use 0.5ml of Histopaque 10831. You will need to use a microcentrifuge tube, preferably a 1.5ml microcentrifuge tube. If the mouse blood is diluted 1:1 with normal salaine or PBS, use a 2.0ml microcentrifuge tube. With such small volumes of blood, it might not be possible to actually see cells at the blood/Histopaque interface/boundary. The interface will simply need to be collected and then spun down. The number of cells collected will not be large.
10831
Histopaque®-1083
sterile-filtered, density: 1.083
Histopaque®-1083
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Product Name
Histopaque®-1083, sterile-filtered, density: 1.083
sterility
sterile-filtered
form
liquid
shelf life
Expiry date on the label.
technique(s)
cell culture | mammalian: suitable
impurities
endotoxin, tested
density
1.083
application(s)
hematology
histology
storage temp.
2-8°C
Quality Level
1 of 4
This Item | 10771 | A6929 | 11191 |
|---|---|---|---|
| form liquid | form liquid | form solution | form liquid |
| density 1.083 | density 1.077 | density 1.077 g/mL | density 1.119 |
| technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable |
| sterility sterile-filtered | sterility sterile-filtered | sterility aseptically filled | sterility sterile-filtered |
| 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 Expiry date on the label. | shelf life Expiry date on the label. |
Application
Histopaque®-1083 has been used for the isolation of mononuclear cells and leukocytes from rat peripheral blood.
Biochem/physiol Actions
Anticoagulated blood is layered onto HISTOPAQUE1083. During centrifugation, erythrocytes are aggregated by polysucrose and rapidly sediment; whereas, lymphocytes and other mononuclear cells remain at the plasma-HISTOPAQUE 1083 interface. Erythrocytes pellet to the bottom of the centrifuge tube.
Features and Benefits
Histopaque®-1083 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
General description
Histopaque®-1083 is a solution containing polysucrose and sodium diatrizoate, adjusted to a relative density of 1.083 g/mL. It facilitates the recovery of viable mononuclear cells from small volumes of blood of rats, mice, and other small mammals.
Legal Information
Histopaque is a registered trademark of Merck KGaA, Darmstadt, Germany
signalword
Warning
hcodes
Hazard Classifications
Eye Irrit. 2 - Met. Corr. 1 - Skin Irrit. 2
Storage Class
8A - Combustible corrosive hazardous materials
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Faceshields, Gloves, type ABEK (EN14387) respirator filter
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This troubleshooting guide addresses the most common sources of error observed when using Histopaque but is not meant to be a comprehensive list.
Related Content
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How much histopaque should we use for 0.5 mL of mouse blood?
1 answer-
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Could less than 3ml of mouse blood (undiluted or diluted) be used with histopaque 1083? If so, what is the recommended minimum volume and should the volume of histopaque also be adjusted?
1 answer-
Yes. It is possible to use less than 3 ml of undiluted or diluted mouse blood. The procedure is scalable. But as the blood volume is reduced, the size of the centrifuge tube must be reduced accordingly. The same ratio of blood (undiluted or diluted) should be used compared to the amount of Histopaque used.
Some users have reported that the procedure can be performed in microcentrifuge tubes. No studies have been performed to determine the minimal amount of blood that can be processed. If the volume of blood is less than 1ml, it might be possible to view the mononuclear cell band - or not. The most common sizes of microcentrifuge tubes are 1.5ml and 2.0ml. There are also larger 5ml sizes. If using a 2 ml microcentrifuge tube, would suggest 750-800 microliters of blood to 750-800 microliters of Histopaque 1083 or other appropriate density media.
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Which type of Histopaque should be used to obtain peripheral blood mononuclear cells (PBMCs) from mouse or rat blood?
1 answer-
Histopaque 10831 is formulated for use with rats, mice, and other small animals. Histopaque 10831 is suitable for isolating peripheral blood mononuclear cells (PBMCs) from mouse or rat blood, including bone marrow. Ficoll Paque 1.084 g/ml is another product designed for rats and mice.
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Which density gradient solution would be suitable for separating mouse mononuclear cells, specifically from their lung and spleen? Can either of the mentioned products (10831 or GE17-5446-02) serve this purpose, or do you have any recommendations?
1 answer-
Histopaque 10831 and Ficoll Paque 1.084 are readily formulated and re suitable density gradients for purifying mouse mononuclear cells from blood or tissue. Alternatively, Percoll or Optiprep can also be used for similar purposes.
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What could be the reason for the poor performance, specifically low yields, when using Histopaque 10831 with mouse blood for flow cytometry?
1 answer-
The poor performance in terms of low yields observed when using Histopaque 10831 with mouse blood for flow cytometry might be attributed to a few factors. The factors affecting the yield include insufficient EDTA, improper blood collection technique, using oversized tubes, the presence of low WBC counts, and the mice being on a high-fat diet. By addressing these factors, the yield can potentially be improved.
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What could be the reason for the low yield of peripheral blood mononuclear cells (PBMCs) obtained from rat blood using Histopaque 1083?
1 answer-
There are various reasons for low PBMC yields. In rats, lymphocytes should be the predominant cell type in blood, unlike in humans where neutrophils are predominant. If the cell band is visible, collecting PBMCs should not be difficult. If the first pellet is larger than subsequent pellets, it's possible that the centrifuge tubes carry a positive charge. Improper blood collection methods in rats and mice lead to low yields, e.g., hemolysis from tail vein blood collection. Proper anticoagulant use is crucial. Temperature, centrifuge state, and PBS isotonicity also impact yields. Contaminated PBS or an old, open Histopaque bottle affect recovery. Also, special diets or medications given to the rat may impact recovery in rats.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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I find mononuclear cells separate well from human blood over Product 10831, Histopaque®-1083, but the cell recovery from rats and mice is lower. Why?
1 answer-
The most common cause for separation problems with Histopaque® 1083 is the blood source. Blood from rats and mice is harder to obtain than blood from a human. Most users experiencing problems are collecting blood with a syringe. It is relatively uncommon for users to call in about poor separation when using vacuum tubes. Humanely sacrifice the animal if possible and immediately collect the blood from the aorta or vena cava. Collecting blood from a severed head is never acceptable. Blood obtained from bleeding the tail is also generally unacceptable. Tissue fluids and other extraneous debris will often result in an activation of the coagulation cascase. Blood drawn into a vacuum tube is the best source of blood. See the product insert for Histopaque® 1083 for the amount of anticoagulant to add to blood collected with a syringe.
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Are there limitations when attempting to use Product 10831, Histopaque®-1083 with the various Accuspin products?
1 answer-
Yes. There are some limitations. Small amounts of blood cannot be processed in the Accuspin tubes. At least 3 mls of whole peripheral blood is expected. Without sufficient red cell numbers, the mononuclear band fails to adequately clear the barrier or frit found within the Accuspin tubes.
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Can Product 10831, Histopaque®-1083, be used with human blood?
1 answer-
Cells can be recovered from human blood using Histopaque® 1083. However Sigma sells Histopaque® 1077 specifically for use with human blood. Histopaque® 1077 is intended for In Vitro Diagnostic Use. Histopaque® 1083 is intended for research use only with animal blood.
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