Antibody clone performance following fixation/permeabilization

It is sometimes possible to stain cells after fixation but this is dependent upon the effects of fixation on the epitope of your protein-of-interest. The list below contains information about the performance of different clones after fixation using our Invitrogen™ eBioscience™ Intracellular Fixation & Permeabilization Buffer Set (cat. no. 88-8824) or after IC Fixation Buffer (cat. no. 00-8222) followed by treatment with methanol. Please note that staining after fixation may allow for intracellular stores of protein to also be stained with the antibody.

Antibody clone performance following fixation/permeabilization

AntigenCloneLive cells (before IC fixation)1After IC fixation and perm wash1After IC fixation/ methanol2
HUMAN    
CD3OKT3  +++++ +
 UCHT1 ++++++ +++
 SK7 +++++ +++
CD4OKT4 ++++ +/-
 RPA-T4 ++++++ +
 SK3 +++ +++ +
CD5UCHT2 ++++++ +++
CD8OKT8 ++++/-  - 
 RPA-T8 ++++++ +/-
 SK1 ++++++ +/-
CD8bSIDI8BEE +++++ ++
CD11aHI111 ++++++ +
CD11bCBRM1/5 + - +++
 ICRF44 ++ - ++
CD11c3.9 ++  + ++
CD1461D3 +++ +++ ++
CD19HIB19 +++ ++ +/-
CD202H7 +++ +++ -
CD25BC96 + - +/-
CD27O323 +++  + +
CD31WM59 +++ ++ +++
CD33p67.6 +++ +++ +++
CD38HB7 ++ - ++
 HIT2 ++ - ++
CD405C3 ++ - ++
CD44IM7 +++ +++ +++
CD452D1 +++ +++ +++
 HI30 +++ +++ +++
CD45RAHI100 +++ +++ +++
CD45ROUCHL1 +++ +++ ++
CD56MEM-188 + - +/-
 CB56 + - +/-
 CMSSB ++ +/- -
CD57TB01 ++ ++ ++
CD62LDREG-56 ++ - ++
CD69FN50 +++ ++ ++
CD802D10.4 + - +
CD83HB15e ++ ++ ++
CD86IT2.2 + + +
CD94HP-3D9 ++ ++ ++
CD95DX2 + - +
CD127RDR5 ++ + ++
CD161HP-3G10 ++ ++ +
MOUSE    
CD3145-2C11++ ++/-
 500A2+++ ++++++
 17A2+++ ++++++
CD4GK1.5+++ +++++
 RM4-5+++ ++++++
CD853-6.7+++ ++++
CD11bM1/70++ ++++
CD11cN418+++ +++++
CD191D3+++ ++-
 MB19-1++ --
CD24M1/69+++ +++++
CD25PC61.5+ -+
 3C7++ -+
 7D4++ +++
CD3924DMS1+ -++
CD44IM7++ ++++
CD4530-F11+++ ++++++
CD45R(B220)RA3-6B2+++ +++++
CD49bDX5++ -+
CD69H1.2F3++ --
CD185(CXCR5)SPRCL5++ --
GR-1RB6-8C5++++++++
NK1.1PK136++++
IgD11-26c++++++*++
IgMll/41++++++
TCR betaH57-597++++++
RAT    
CD25OX39 + - 
CANINE    
CD4ykix302.9+++ + 
CD5ykix322.3+++ - 
CD8aycate55.9+++ ++ 
CD44ykix337.8+++ ++ 
CD45ykix716.13+++ ++ 
CD45Rykix753.22+++ ++ 
CD90ykix337.217+++ ++ 
MHC Class IIykix334.2+++ +++ 

1Testing carried out with eBioscience Fixation & Permeabilization Kit for 15–30 minutes at room temperature followed by a perm wash, see Staining Intracellular Antigens for Flow Cytometry, protocol A.

*Please note the clones with * indicate longer fixations are detrimental to staining.

2 Testing carried out for 15–30 minutes at room temperature in 50% IC fixation followed by 30 minutes 90–100% methanol incubation on ice, see Staining Intracellular Antigens for Flow Cytometry, protocol C