CD117 Nanobeads, mouse (RUO)

Catalog Number: S0K0031 Reactivity: Mouse Conjugation: Brand: Starter

Price:

$550 USD

Size: 1 mL 2 mL

Quantity:

For shipping services or bulk orders, you may request a quotation.

Secure checkout with

View full details

Product Details

Product Specification

Format	Dextran-coated iron oxide nanomagnetic beads coupled with an anti-mouse CD117 monoclonal antibody are stored in a PBS buffer system containing BSA, Poloxamer 188, and NaN.₃
Capacity	1 mL magnetic beads: suitable for 1 × 10⁹Total cell count: up to 100 sorting runs can be performed. 2 mL magnetic beads: suitable for 2 × 10⁹Total cell count: up to 200 sorting runs can be performed.
Principle of Separation	First, CD117-positive cells are magnetically labeled using CD117 Nanobeads, mouse. After labeling, the cell suspension is loaded into a separation column placed within the sorter's magnetic field. The magnetically labeled CD117⁺ cells are retained in the column, while the unlabeled cells flow through. At this point, the cells that elute from the column constitute the negative fraction after removal of CD117⁺ cells. Once the column is removed from the magnetic field, the CD117⁺ cells captured by the magnetic beads are eluted, yielding the positive fraction.
Applications in cell sorting	Mouse cells enriched for or depleted of CD117+ cells
Reactivity	Mouse
Magnetic Bead Size	20 nm
Separation Method	Column-Based
Selection Strategy	Positive Selection
Stability & Storage	Store protected from light at 2–8°C; avoid freezing.

Background

Mouse CD117, often also referred to as c-Kit or the stem cell factor receptor (SCFR), is a crucial type III receptor tyrosine kinase. It is expressed on most hematopoietic progenitor cells, including multipotent hematopoietic stem cells as well as differentiated myeloid, erythroid, and lymphoid progenitors. It has been reported that, in addition to their hematopoietic differentiation potential, CD117-positive stem cells in mouse bone marrow can also differentiate into smooth muscle cells, muscle cells, and endothelial cells in vivo. CD117 is also expressed on certain mature hematopoietic cells, such as mast cells.

Protocol

Note: This kit does not include the sorting buffer; it must be provided separately. The corresponding Stater product number is S0D3018.

Taking SSeparation Columnas an example

	Steps	Operating Procedure	Dosage / Time
Magnetic Labeling	1	Take a mouse tissue sample containing CD117⁺cells, grind it on a 40 μm sterile filter to prepare a single-cell suspension; after red blood cell lysis, perform live cell counting. Note:It is recommended to count cells after staining with AO/PI.	—
	2	Centrifuge the cell suspension at 400×g for 7 minutes, and completely discard the supernatant.	400×g, 7 min
	3	Resuspend every 1 × 10⁷ cells in 90 μL of buffer. Note:Adjust the volume of buffer proportionally according to the sample volume; (e.g., resuspend 2 × 10⁷ cells in 180 μL of sorting buffer).	Resuspend the cells.
	4	Add 10 μL per 1 × 10⁷ cells.CD117 Nanobeads, mouse,And mix the cell suspension thoroughly, avoiding excessive bubble formation during mixing. Note:When the cell number increases, increase the volume of all reagents proportionally; (e.g., for 2 × 10⁷ cells, add 20 μL of sorting magnetic beads.)	For 1 × 10⁷ cells, add 10 μL.CD117 Nanobeads, mouse
	5	Mix thoroughly and incubate in the dark at 2–8°C for 15 minutes.	Incubate for 15 minutes.
	6	Add 1 mL of buffer per 1 × 10⁷ cells to wash, centrifuge at 400 × g for 7 minutes, and completely discard the supernatant.	Add 1 mL of buffer for 1 × 10⁷ cells.
	7	Resuspend the cells in 500 μL of sorting buffer. Note:500 μL of buffer can resuspend up to 1 × 10⁸ cells.Cells	Resuspend the cells.
Magnetic separation.	8	Place the S-shaped separation column into the magnetic field of the corresponding magnetic separator, and rinse the column with 500 μL of buffer.	Rinse the separation column.
	9	Load the cell suspension onto the separation column and collect the flow-through (unlabeled cells).	—
	10	Wash the column with buffer three times, each time using 500 μL; add the next wash only after the liquid in the column has completely drained.	3 × 500 μL of buffer.
	11	Combine all flow-through fractions as the negative fraction; remove the separation column from the magnetic field and place it over a collection tube.	—
	12	Add 1 mL of buffer to the column, then quickly and firmly push down with the provided plunger to elute the magnetically labeled positive cells. Note:For higher purity, use a new column and repeat the separation once.	Collect the target cells.

In LTaking the Separation Columnas an example.

	Steps	Operating Procedure	Dosage / Time
Magnetic Labeling	1	Take CD117⁺ containingMouse tissue samples are ground on a 40 μm sterile filter to prepare a single-cell suspension; after red blood cell lysis, live cell counting is performed. Note: It is recommended to count cells after staining with AO/PI.	—
	2	Centrifuge the cell suspension at 400×g for 7 minutes, and completely discard the supernatant.	400×g, 7 min
	3	Resuspend every 1 × 10⁷ cells in 90 μL of buffer. Note:Adjust the volume of buffer proportionally according to the sample volume; (e.g., resuspend 2 × 10⁷ cells in 180 μL of sorting buffer).	Resuspend the cells
	4	Add 10 μL per 1 × 10⁷ cellsCD117 Nanobeads, mouse,and mix the cell suspension thoroughly, avoiding excessive bubble formation during mixing. Note:When the cell number increases, all reagent volumes increase proportionally; (e.g., for 2×10⁷ cells, add 20 μL of sorting magnetic beads)	For 1×10⁷ cells, add 10 μLCD117 Nanobeads, mouse
	5	Mix thoroughly and incubate in the dark at 2–8°C for 15 minutes	Incubate for 15 min
	6	Wash with 1 mL of buffer, centrifuge at 400×g for 7 minutes, and completely discard the supernatant	For 1×10⁷ cells, add 1 mL of buffer
	7	Resuspend the cells in 1 mL of sorting buffer Note:A maximum of 1×10⁸ cells can be resuspended in 500 μL of bufferCells	Resuspend cells
Magnetic separation	8	Place the L-shaped separation column into the magnetic field of the corresponding magnetic separator, and rinse the column with 3 mL of buffer	Rinse the separation column
	9	Load the cell suspension onto the separation column and collect the flow-through (unlabeled cells)	—
	10	Wash the column with buffer 3 times, 3 mL each time, and add the next wash only after the liquid in the column has completely drained.	3 × 3 mL of buffer
	11	Combine all eluates into the negative fraction; remove the separation column from the magnetic field and place it over a collection tube.	—
	12	Add 5 mL of buffer to the column, then quickly and firmly push down with the provided plunger to elute the magnetically labeled positive cells. Note:If higher purity is required, use a new column and repeat the separation once.	Collect the target cells.

Item added to your cart

CD117 Nanobeads, mouse (RUO)