Shopping Days 開催中 | 詳細はこちら >>
43481
FoxP3/Transcription Factor Fixation/Permeabilization Kit
Buffers & Dyes

FoxP3/Transcription Factor Fixation/Permeabilization Kit #43481

現在発売の準備をしております。次回新発売までお待ちください。

Flow Cytometry Image Product Gallery Navigation1

Flow cytometric analysis of mouse splenocytes fixed and permeabilized with #43481 FoxP3/Transcription Factor Fixation/Permeabilization kit, using FoxP3 (3G3) Mouse mAb (PE Conjugate) #49402 (right) and co-stained with CD4-APC #78292, compared to a concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (PE Conjugate) #63630 (left).

Learn more about how we get our images

FoxP3/Transcription Factor Fixation/Permeabilization Kit #43481

A. Solutions and Reagents

NOTE: Precipitation may occur in some kit components. The presence of precipitate does not affect the performance of the reagent.

Supplied Reagents:

  • FoxP3/Transcription Factor Fixation/Permeabilization Diluent (1X) (#58766)
  • FoxP3/Transcription Factor Fixation/Permeabilization Concentrate (4X) (#44931): Dilute desired amount to a 1X working solution with FoxP3/Transcription Factor Fixation/Permeabilization Diluent (1X)
  • FoxP3/Transcription Factor Permeabilization Buffer (10X) (#68751): Dilute desired amount to a 1X working solution with reverse osmosis deionized (RODI) or equivalent grade water. Use for all wash steps and antibody incubation following fixation.

Additional Reagents (Not Supplied):

  • Red blood cell lysis buffer
  • Unconjugated or fluorochrome-conjugated primary antibodies
  • Fluorochrome-conjugated secondary antibodies (if applicable)
  • Cellular dyes (e.g., live-dead discrimination, DNA dyes, etc.), optional. Visit www.cellsignal.com/flowdyes for a full listing of flow cytometry validated cellular dyes.

B. Fixation and Permeabilization

NOTE: Fixable viability dyes such as Ghost Dye™ Violet 510 Viability Dye #59863 should be added prior to fixation, following the dye product recommended protocol. Proceed with fixation once excess dye has been removed.

NOTE: Add antibodies targeting CD markers or other extracellular epitopes prior to fixation. The antibodies will remain bound to the target of interest during the fixation process. A wash step prior to fixation may be performed but is not necessary.

  1. Pellet cells by centrifugation and aspirate supernatant.
  2. Resuspend cells in 1 mL FoxP3/Transcription Factor Fixation/Permeabilization 1X working solution, prepared as described above. Mix well to dissociate pellet and prevent cross-linking of individual cells.
  3. Incubate 30-60 min at room temperature (20-25 ºC). Protect from light.
  4. Wash cells 2 times in excess 1X FoxP3/Transcription Factor Permeabilization Buffer.

C. Immunostaining

  1. Aliquot desired number of cells into tubes or wells.
  2. Resuspend cells in 100 µl of diluted antibody, prepared in 1X FoxP3/Transcription Factor Permeabilization Buffer at the recommended dilutions. See individual antibody datasheets or product webpage for recommended dilution, or determine via titration.
  3. Incubate for 1 hr at room temperature (20-25 °C). Protect from light.
  4. Wash by centrifugation in excess 1X FoxP3/Transcription Factor Permeabilization Buffer. Discard supernatant. Repeat.
  5. If using a fluorochrome-conjugated primary antibody, resuspend cells in 500 µl 1X FoxP3/Transcription Factor Permeabilization Buffer and analyze on flow cytometer; for unconjugated primary antibodies, proceed to next step.
  6. Resuspend cells in fluorochrome-conjugated secondary antibody, diluted in 1X FoxP3/Transcription Factor Permeabilization Buffer at the recommended dilution.
  7. Incubate for 30 min at room temperature (20-25 °C). Protect from light.
  8. Wash by centrifugation in excess 1X FoxP3/Transcription Factor Permeabilization Buffer. Discard supernatant. Repeat.
  9. Resuspend cells in 500 µl 1X FoxP3/Transcription Factor Permeabilization Buffer and analyze on flow cytometer.
Product Includes Quantity
FoxP3/Transcription Factor Fixation/Permeabilization Concentrate (4X) 50 ml
FoxP3/Transcription Factor Fixation/Permeabilization Diluent (1X) 160 ml
FoxP3/Transcription Factor Permeabilization Buffer (10X) 150 ml

This kit provides the reagents needed to support antibody-based detection of FoxP3 and other transcription factors by flow cytometry. Optimally formulated fixation and permeabilization buffers halt biological activity and enable antibody access to intracellular targets. Antibody dilution, incubation, and wash steps are performed with the included FoxP3/Transcription Factor Permeabilization Buffer, eliminating the need for separate reagents.

Note: Precipitation may occur in some kit components. The presence of precipitate does not affect the performance of the reagent.

Storage:

Store at 4ºC. All components in this kit are stable for at least 6 months when stored at 4ºC.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

News from the Bench

Discover what’s going on at CST, receive our latest application notes and tips, read our science features, and learn about our products.

Subscribe

To Purchase # 43481S

View sizes
Product # Size Price Inventory
43481S
1 Kit

Your Local Representative for Japan

Cell Signaling Technology Japan, K.K.

10F Kasahara Building, 1-6-10 Uchikanda

Chiyoda-ku, Tokyo 101-0047

Phone:
03 (3295) 1630
Fax:
03 (3295) 1633
Email:
infojp@cellsignal.com
URL:
www.cstj.co.jp

Need information for a different country? Please click here.

To get local purchase information on this product, click here.