Loading...

Cyto-Fast™ Fix/Perm Buffer Set Cyto-Fast™ Fix/Perm Buffer Set

Pricing & Availability
Regulatory Status
RUO
Other Names
CytoFast
Ave. Rating
Submit a Review
Product Citations
publications
Cyto-Fast_Fix_Perm_Buffer_Set_110718
PMA + ionomycin stimulated (3 hours) human peripheral blood lymphocytes (in the presence of monensin) were fixed and permeabilized with Cyto-Fast™ Fix/Perm Buffer Set. Cells were then stained with CD3 (UCHT1) APC and isotype control PE (left panel), IL-2 (clone MQ1-17H12) PE (middle panel), or IFN-γ (clone 4S.B3) PE (right panel).
  • Cyto-Fast_Fix_Perm_Buffer_Set_110718
    PMA + ionomycin stimulated (3 hours) human peripheral blood lymphocytes (in the presence of monensin) were fixed and permeabilized with Cyto-Fast™ Fix/Perm Buffer Set. Cells were then stained with CD3 (UCHT1) APC and isotype control PE (left panel), IL-2 (clone MQ1-17H12) PE (middle panel), or IFN-γ (clone 4S.B3) PE (right panel).
Cat # Size Price Quantity Check Availability Save
426803 300 tests DKK1640
Check Availability


Need larger quantities of this item?
Request Bulk Quote
Description

Cyto-Fast™ Fix/Perm Buffer Set is composed of Cyto-Fast™ Fix/Perm Solution and Cyto-Fast™ Perm Wash Solution (10X). It is designed for fixation and permeabilization of mammalian cells for intracellular staining such as cytokines and other cytoplasmic molecules.

Product Details
Technical Data Sheet (pdf)

Product Details

Storage & Handling
Upon receipt, store at 2-8°C.
Application

ICFC - Quality tested
Recommended Usage

Procedure for staining in round-bottom 5mL tubes:

  1. Dilute the Cyto-Fast™ Perm Wash solution (10X) to 1X using deionized water
  2. Aliquot 100 µL of cells (2 x 105-1 x 106) of interest into a 12 x 75 mm tube 
  3. Add 150 µL of Cyto-Fast™ Fix/Perm Solution and mix
  4. Incubate for 20 minutes at room temperature
  5. Add 1 mL of 1X Cyto-Fast™ Perm Wash solution
  6. Centrifuge at 350 xg for 5 minutes, discard supernatant
  7. Repeat steps 5 - 6
  8. Stain cells with optimal concentration of intracellular antibodies. Prepare antibodies in 1X Cyto-Fast™ Perm Wash Solution, in 100 µL total volume
  9. Incubate 20 minutes at room temperature in the dark
  10. Wash cells with 1 mL of 1x Cyto-Fast™ Perm Wash solution
  11. Centrifuge at 350 xg for 5 minutes, discard supernatant
  12. Add 1 mL of Cell Staining Buffer (Cat. No. 420201)
  13. Centrifuge at 350 xg for 5 minutes, discard supernatant
  14. Resuspend the cells in 300 µL of Cell Staining Buffer
  15. Acquire samples on a flow cytometer.

Procedure for staining in 96-well U-bottom plates:

  1. Dilute the Cyto-Fast™ Perm Wash solution (10X) to 1X using deionized water
  2. Aliquot 100 µL of cells (2 x 105-1 x 106) of interest into individual wells in a 96-well U-bottom plate 
  3. Centrifuge at 350 xg for 5 minutes, discard supernatant, and loosen cell pellet.
  4.  Add 100 µL of Cyto-Fast™ Fix/Perm Solution and mix
  5. Incubate for 20 minutes at room temperature
  6. Add 100 µL of 1X Cyto-Fast™ Perm Wash solution
  7. Centrifuge at 350 xg for 5 minutes, discard supernatant
  8. Repeat steps 6 - 7 by adding 200 µL of 1X Cyto-Fast™ Perm Wash solution
  9. Stain cells with optimal concentration of intracellular antibodies. Prepare antibodies in 1X Cyto-Fast™ Perm Wash Solution, in 100 µL total volume
  10. Incubate 20 minutes at room temperature in the dark
  11. Wash cells by adding 100 µL of 1x Cyto-Fast™ Perm Wash solution
  12. Centrifuge at 350 xg for 5 minutes, discard supernatant
  13. Add 200 µL of Cell Staining Buffer (Cat. No. 420201)
  14. Centrifuge at 350 xg for 5 minutes, discard supernatant
  15. Resuspend the cells in 200 µL of Cell Staining Buffer
  16. Acquire samples on a flow cytometer.

Addional Notes

  • Cells can be bulk fixed. To bulk fix the cells, add 1.5 mL of Cyto-Fast™ Fix/Perm Solution for each 1 x 107 cells and incubate for 15-20 minutes at room temperature.
  • Staining for surface markers can be performed prior to fix/perm. Follow the Cell Surface Flow Cytometry Staining Protocol. Proceed to follow the Cyto-Fast™ Fix/Perm Buffer Set protocol outlined above after primary (and/or secondary) surface antibody staining steps.
  • If cells are to be immediately stained post-fix/perm, wash cells 2 x with 10 mL of 1X Cyto-Fast™ Perm Wash Solution. Aliquot 100µL of cells (2 x 105-1 x 106) of interest into a 12 x 75 mm tube and stain cells with optimal concentration of antibodies (continue with procedure listed above at step 9).
  • If cells are to be stained at a later time post-fix/perm, wash cells with 10 mL of Cell Staining Buffer. Cells can be stored for up to one week in Cyto-Last™ Buffer (Cat. No. 422501).
  • To stain cryopreserved cells, quickly thaw the cells by placing them in a 37°C water bath, wash with 10 mL of 1X Cyto-Fast™ Perm Wash Solution and centrifuge for 5 minutes at 200-300xg, discard supernatant. Re-suspend cells in 1X Cyto-Fast™ Perm Wash Solution for 15-20 minutes prior to staining.
  • The Cyto-Fast™ Fix/Perm Buffer set (10x) may have crystallization or precipitation when it is stored at 2-8°C this is normal and does not affect the buffer performance. If heavy precipitation is observed after  diluting to a 1X working solution, it can be filtered to clarify the solution.
Application Notes

Cyto-Fast™ Fix/Perm Buffer Set Contents:

  • Cyto-Fast™ Fix/Perm Solution (50 mL)
  • Cyto-Fast™ Perm Wash Solution 10X (100 mL)

Antigen Details

Molecular Family
Cytokines/Chemokines
Gene ID
NA

Related FAQs

There are no FAQs for this product.
Go To Top Version: 3    Revision Date: 10/21/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

Pricing & Availability
Regulatory Status
RUO
Other Names
CytoFast
Ave. Rating
Submit a Review
Product Citations
publications
Cyto-Fast_Fix_Perm_Buffer_Set_110718
PMA + ionomycin stimulated (3 hours) human peripheral blood lymphocytes (in the presence of monensin) were fixed and permeabilized with Cyto-Fast™ Fix/Perm Buffer Set. Cells were then stained with CD3 (UCHT1) APC and isotype control PE (left panel), IL-2 (clone MQ1-17H12) PE (middle panel), or IFN-γ (clone 4S.B3) PE (right panel).
  • Cyto-Fast_Fix_Perm_Buffer_Set_110718
    PMA + ionomycin stimulated (3 hours) human peripheral blood lymphocytes (in the presence of monensin) were fixed and permeabilized with Cyto-Fast™ Fix/Perm Buffer Set. Cells were then stained with CD3 (UCHT1) APC and isotype control PE (left panel), IL-2 (clone MQ1-17H12) PE (middle panel), or IFN-γ (clone 4S.B3) PE (right panel).
Cat # Size Price Quantity Check Availability Save
426803 300 tests DKK1640
Check Availability


Need larger quantities of this item?
Request Bulk Quote
Description

Cyto-Fast™ Fix/Perm Buffer Set is composed of Cyto-Fast™ Fix/Perm Solution and Cyto-Fast™ Perm Wash Solution (10X). It is designed for fixation and permeabilization of mammalian cells for intracellular staining such as cytokines and other cytoplasmic molecules.

Product Details
Technical Data Sheet (pdf)

Product Details

Storage & Handling
Upon receipt, store at 2-8°C.
Application

ICFC - Quality tested
Recommended Usage

Procedure for staining in round-bottom 5mL tubes:

  1. Dilute the Cyto-Fast™ Perm Wash solution (10X) to 1X using deionized water
  2. Aliquot 100 µL of cells (2 x 105-1 x 106) of interest into a 12 x 75 mm tube 
  3. Add 150 µL of Cyto-Fast™ Fix/Perm Solution and mix
  4. Incubate for 20 minutes at room temperature
  5. Add 1 mL of 1X Cyto-Fast™ Perm Wash solution
  6. Centrifuge at 350 xg for 5 minutes, discard supernatant
  7. Repeat steps 5 - 6
  8. Stain cells with optimal concentration of intracellular antibodies. Prepare antibodies in 1X Cyto-Fast™ Perm Wash Solution, in 100 µL total volume
  9. Incubate 20 minutes at room temperature in the dark
  10. Wash cells with 1 mL of 1x Cyto-Fast™ Perm Wash solution
  11. Centrifuge at 350 xg for 5 minutes, discard supernatant
  12. Add 1 mL of Cell Staining Buffer (Cat. No. 420201)
  13. Centrifuge at 350 xg for 5 minutes, discard supernatant
  14. Resuspend the cells in 300 µL of Cell Staining Buffer
  15. Acquire samples on a flow cytometer.

Procedure for staining in 96-well U-bottom plates:

  1. Dilute the Cyto-Fast™ Perm Wash solution (10X) to 1X using deionized water
  2. Aliquot 100 µL of cells (2 x 105-1 x 106) of interest into individual wells in a 96-well U-bottom plate 
  3. Centrifuge at 350 xg for 5 minutes, discard supernatant, and loosen cell pellet.
  4.  Add 100 µL of Cyto-Fast™ Fix/Perm Solution and mix
  5. Incubate for 20 minutes at room temperature
  6. Add 100 µL of 1X Cyto-Fast™ Perm Wash solution
  7. Centrifuge at 350 xg for 5 minutes, discard supernatant
  8. Repeat steps 6 - 7 by adding 200 µL of 1X Cyto-Fast™ Perm Wash solution
  9. Stain cells with optimal concentration of intracellular antibodies. Prepare antibodies in 1X Cyto-Fast™ Perm Wash Solution, in 100 µL total volume
  10. Incubate 20 minutes at room temperature in the dark
  11. Wash cells by adding 100 µL of 1x Cyto-Fast™ Perm Wash solution
  12. Centrifuge at 350 xg for 5 minutes, discard supernatant
  13. Add 200 µL of Cell Staining Buffer (Cat. No. 420201)
  14. Centrifuge at 350 xg for 5 minutes, discard supernatant
  15. Resuspend the cells in 200 µL of Cell Staining Buffer
  16. Acquire samples on a flow cytometer.

Addional Notes

  • Cells can be bulk fixed. To bulk fix the cells, add 1.5 mL of Cyto-Fast™ Fix/Perm Solution for each 1 x 107 cells and incubate for 15-20 minutes at room temperature.
  • Staining for surface markers can be performed prior to fix/perm. Follow the Cell Surface Flow Cytometry Staining Protocol. Proceed to follow the Cyto-Fast™ Fix/Perm Buffer Set protocol outlined above after primary (and/or secondary) surface antibody staining steps.
  • If cells are to be immediately stained post-fix/perm, wash cells 2 x with 10 mL of 1X Cyto-Fast™ Perm Wash Solution. Aliquot 100µL of cells (2 x 105-1 x 106) of interest into a 12 x 75 mm tube and stain cells with optimal concentration of antibodies (continue with procedure listed above at step 9).
  • If cells are to be stained at a later time post-fix/perm, wash cells with 10 mL of Cell Staining Buffer. Cells can be stored for up to one week in Cyto-Last™ Buffer (Cat. No. 422501).
  • To stain cryopreserved cells, quickly thaw the cells by placing them in a 37°C water bath, wash with 10 mL of 1X Cyto-Fast™ Perm Wash Solution and centrifuge for 5 minutes at 200-300xg, discard supernatant. Re-suspend cells in 1X Cyto-Fast™ Perm Wash Solution for 15-20 minutes prior to staining.
  • The Cyto-Fast™ Fix/Perm Buffer set (10x) may have crystallization or precipitation when it is stored at 2-8°C this is normal and does not affect the buffer performance. If heavy precipitation is observed after  diluting to a 1X working solution, it can be filtered to clarify the solution.
Application Notes

Cyto-Fast™ Fix/Perm Buffer Set Contents:

  • Cyto-Fast™ Fix/Perm Solution (50 mL)
  • Cyto-Fast™ Perm Wash Solution 10X (100 mL)

Antigen Details

Molecular Family
Cytokines/Chemokines
Gene ID
NA
Go To Top Version: 3    Revision Date: 10/21/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

ProductsHere
Login / Register
Remember me
Forgot your password? Reset password?
Create an Account