Rat and Mouse Heart Tissue Single-cell Preparation Kit servicebio G4152-50T
$50.00
In stock
SKU
G4152-50T
Product Information
| Product Name: Rat and Mouse Heart Tissue Single-cell Preparation Kit | Cat. No.: G4152-50T |
| Spec.: 50T |
Product Description / Introduction
This product is suitable for dissociating rat and mouse Heart tissue into single-cell suspensions, it is used in conjunction with a grinding instrument and grinding beads (zirconia), Standardized processing organization, the dissociation speed is fast, the survival rate of dissociated single cells reaches over 80 %, the cell agglomeration rate is less than 30 %, the extracted single cells can be subjected to flow cytometry, Single-cell sequencing and other experiments.
Storage and Shipping Conditions
-20 ℃ Storage, Dry ice transportation, Valid for 12 months.
Product Content
| Component Number: G4152-1 | Component: Rat and Mouse Heart tissue dissociation reagent B |
| G4152-50T: 25mg | Component Number: G4152-2 |
| Component: Rat and Mouse Heart tissue dissociation reagent E | G4152-50T: 62.5mg |
| Product Manuals | 1 copy |
Assay Protocol / Procedures
1. Preparations before the experiment.
a) Transfer 1 mL of Dissociation Reagent B into HBSS or basal medium containing Ca2+/Mg2+ (G4204) , shake to dissolve. Transfer the solution to a 50 mL centrifuge tube. Rinse the reagent container with additional HBSS or basal medium containing Ca2+/Mg2+ (G4204) , and combine the rinse solution into the same 50 mL centrifuge tube. Adjust the final volume to 25 mL, sterilize by filtration through a 0.22 μm filter, and aliquot for later use.
b) Transfer 1 mL of Dissociation Reagent E into HBSS or basal medium containing Ca2+/Mg2+ (G4204) , shake to dissolve. Transfer the solution to a 50 mL centrifuge tube. Rinse the reagent container with additional HBSS or basal medium containing Ca2+/Mg2+ (G4204), and combine the rinse solution into the same 50 mL centrifuge tube. Adjust the final volume to 25 mL, sterilize by filtration through a 0.22 μm filter, and aliquot for later use.
c) Prepare Dissociation Reagent B and Dissociation Reagent E by mixing them at a 1:1 ratio according to experimental requirements. Mix thoroughly until homogeneous and set aside for use.
d) Take a 2 mL grinding tube and add 2 3 mm grinding beads (zirconia) to it for later use.
2. Tissue dissociation.
a) Take the separated tissue (100 mg-200 mg), gently cut it into small pieces and put it into the grinding tube.
e) Add 1 mL of the mixed dissociation reagent solution into the grinding tube.
f) Set the program of the grinding instrument and put the grinding tube in to run.
g) Take the supernatant, filter it through a 200-mesh cell sieve, then centrifuge and remove the supernatant.
h) Wash three times with PBS, remove the supernatant, and the single-cell suspension can be obtained.
3. Reference for operating parameters
The corresponding tissue grinding program has been set inside the instrument. If there are special requirements, the following parameters can be referred to for modification.
| Instrument model: SWE-C6 | Organizational type: Intestine |
| Temperature setting: Room temperature | Grinding time: 8 min |
| Frequency: 25 HZ | Instrument model: SWE-C6 |
| Organizational type: Heart | Temperature setting: Room temperature |
| Grinding time: 10 min | Frequency: 25 HZ |
| Instrument model: SWE-C6 | Organizational type: Liver |
| Temperature setting: Room temperature | Grinding time: 15 min |
| Frequency: 25 HZ | Instrument model: SWE-C6 |
| Organizational type: mouse tumor | Temperature setting: Room temperature |
| Grinding time: 10 min | Frequency: 25 HZ |
| Instrument model: SWE-C6 | Organizational type: lung |
| Temperature setting: Room temperature | Grinding time: 15min |
| Frequency: 25 HZ | Instrument model: SWE-D6 |
| Organizational type: Intestine | Temperature setting: Room temperature |
| Grinding time: 2 min | Frequency: 1 m/s |
| Instrument model: SWE-D6 | Organizational type: Heart |
| Temperature setting: Room temperature | Grinding time: 1 min |
| Frequency: 1 m/s | Instrument model: SWE-D6 |
| Organizational type: Liver | Temperature setting: Room temperature |
| Grinding time: 2 min | Frequency: 1m/s |
| Instrument model: SWE-D6 | Organizational type: mouse tumor |
| Temperature setting: Room temperature | Grinding time: 2 min |
| Frequency: 1 m/s | Instrument model: SWE-D6 |
| Organizational type: lung | Temperature setting: Room temperature |
| Grinding time: 5min | Frequency: 1 m/s |
| Instrument model: SWE-FP | Organizational type: Intestine |
| Temperature setting: 4 ℃ | Grinding time: 3 min |
| Frequency: 25 HZ | Instrument model: SWE-FP |
| Organizational type: Heart | Temperature setting: 4 ℃ |
| Grinding time: 10 min | Frequency: 25 HZ |
| Instrument model: SWE-FP | Organizational type: Liver |
| Temperature setting: 4 ℃ | Grinding time: 15 min |
| Frequency: 25 HZ | Instrument model: SWE-FP |
| Organizational type: mouse tumor | Temperature setting: 4 ℃ |
| Grinding time: 10 min | Frequency: 25 HZ |
| Instrument model: SWE-FP | Organizational type: lung |
| Temperature setting: 4 ℃ | Grinding time: 15 min |
| Frequency: 25 HZ | Instrument model: SWE-3D |
| Organizational type: Intestine | Temperature setting: 4 ℃ |
| Grinding time: 2 min | Frequency: 1 m/s |
| Instrument model: SWE-3D | Organizational type: Heart |
| Temperature setting: 4 ℃ | Grinding time: 2 min |
| Frequency: 1 m/s | Instrument model: SWE-3D |
| Organizational type: Liver | Temperature setting: 4 ℃ |
| Grinding time: 2 min | Frequency: 1 m/s |
| Instrument model: SWE-3D | Organizational type: mouse tumor |
| Temperature setting: 4 ℃ | Grinding time: 2 min |
| Frequency: 1 m/s | Instrument model: SWE-3D |
| Organizational type: lung | Temperature setting: 4 ℃ |
| Grinding time: 5 min | Frequency: 1 m/s |
Note
1. The Dissociation Reagent Kit (50T) can digest a total of 5-10 g of tissue. Excessive tissue may compromise digestion efficiency.
2. The prepared dissociation reagent can be aliquoted and stored at -20 °C protected from light.
3. If single-cell sequencing experiments are to be conducted, sterile and enzyme-free consumables must be selected.
4. For your health, please wear a lab coat and gloves during the experiment.
For Research Use Only!
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