True-Nuclear™ 小鼠 Treg Flow™ 试剂

True-Nuclear™ Mouse Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD4 APC/CD25 PE) 详细如下：

Description ：T regulatory (Treg) cells are a subset of T lymphocytes which is characterized byCD4+/CD25+/FOXP3+. These naturally occurring Treg cells originate in the thymus, andcomprise 2-10% of peripheral CD4+ T cells. It has been shown that Treg cells are able toinhibit T cells proliferation and cytokine production and play critical roles in preventingautoimmunity as well as in controlling tumor immunity and transplantation tolerance. ImpairedTreg function or Treg cell deficiency will develop variety of autoimmune diseases, while higherfrequency of Treg cells will cause hypo-immune response to pathogens.

BioLegend's True-Nuclear™ Mouse Treg Flow™ Kit is designed and formulatedspecifically for immunofluroscence staining and flow cytometric analysis of mouse Treg cells ina mixed lymphocyte population. This kit is composed of fluorochrome conjugated anti-mouseCD4, CD25, FOXP3 antibodies, and the critical buffers. It is easy to use for identification ofTreg cells.

Verified Reactivity ：Mouse

Antibody Type ：Monoclonal

Concentration ：Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in ourCertificate of Analysis online tool.)

Storage & Handling ：This kit is guaranteed for six months. Upon receipt, store between 2°C and 8°C, and protectedfrom prolonged exposure to light. Do not freeze.

Application ：ICFC - Quality tested

Application Notes Materials Provided:
1. Alexa Fluor® 488 anti-mouse FOXP3 - 25 tests
2. Alexa Fluor® 488 Mouse IgG1, ? isotype control - 25 tests
3. True-Nuclear™ Transcription Factor Buffer Set - 120 tests (Cat. No. 424401)
4. anti-mouse CD4 APC/CD25 PE Cocktail - 50 tests

Materials Not Included:
1. Cell Staining Buffer (Cat. No. 420201)
2. Single color compensation controls
Immunofluorescence Staining Procedures:

1. Perform cell surface staining as described in BioLegend's Cell Surface ImmunofluorescenceStaining Protocol. Add 20 µL of the anti-mouse CD4 APC/CD25 PE cocktail to each tube andincubate in the dark for 20 minutes.
2. Add 2 mL of the cell staining buffer, centrifuge tubes at 400 x g at room temperature for fiveminutes, and discard the supernatant.
3. Repeat Step 2, for a total of two washes.
4. Add 1 mL of the Transcription Factor 1X Fix solution to each tube, vortex, and incubate at roomtemperature in the dark for 45-60 minutes.
5. Without washing, add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
6. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
7. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
8. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
9. Resuspend the cell pellet in 100 µl of the Transcription Factor 1XPerm Buffer.
10. Add 5 µL of Alexa Fluor® 488 anti-mouse FOXP3 antibody or 5 µL of Alexa Fluor® 488 mouseIgG1, ? isotype control into the appropriate tubes. Incubate in the dark at room temperature for atleast 30 minutes.
11. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
12. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
13. Add 2 mL of the cell staining buffer.
14. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
15. Resuspend in 0.5 mL cell staining buffer and then acquire tubes on a flow cytometer.

Caution: The True-Nuclear™ Transcription Factor Buffer Set contains para-formaldehyde, which istoxic and mutagenic. Please handle with caution. Wear gloves, lab coats, and necessary protectionto avoid direct contact.

NOTE: For flow cytometric staining with this clone, True-Nuclear™ Transcription Factor Buffer Set(Cat. No. 424401) offers improved staining and is highly recommended over the Foxp3/Perm BufferSet (Cat. No. 421403).