Antibody Fixation Considerations
eBioscience Antibody Clone Performance Following Fixation / Permeabilization
It is possible that fixing cells prior to staining can alter the antibody-binding epitope on a protein. This alteration of the epitope may affect the antibody’s binding affinity. Therefore, we have investigated the effects of fixation and permeabilization using our eBioscience Fixation & Permeabilization Kit (cat. no. 88-8823). The activity of eBioscience antibodies following fixation is summarized in the table below.
| Antigen | Clone | Before | After |
|---|---|---|---|
| HUMAN | |||
| CD3 | OKT3 | +++ | + |
| UCHT1 | +++ | ++ | |
| SK7 | +++ | +++ | |
| CD4 | OKT4 | +++ | + |
| RPA-T4 | +++ | +++ | |
| SK3 | +++ | +++ | |
| CD8 | OKT8 | +++ | +/- |
| RPA-T8 | +++ | +++ | |
| SK1 | +++ | +++ | |
| CD8b | SIDI8BEE | +++ | ++ |
| CD11b | CBRM1/5 | + | - |
| ICRF44 | ++ | +/- | |
| CD11c | 3.9 | ++ | + |
| CD25 | BC96 | + | - |
| CD14 | 61D3 | +++ | ++ |
| CD19 | HIB19 | +++ | ++ |
| SJ25C1 | +++ | ++ | |
| CD20 | 2H7 | +++ | +++ |
| CD25 | CD25-4E3 | ++ | ++ |
| CD45 | 2D1 | +++ | +++ |
| CD45RA | HI100 | ++ | + |
| CD45RO | UCHL1 | ++ | ++ |
| CD62L | DREG-56 | ++ | - |
| CD56 | MEM-188 | + | - |
| CB56 | + | - | |
| CMSSB | ++ |
++ |
|
| MOUSE | |||
| CD3 | 145-2C11 | ++ | + |
| CD4 | RM4-5 | +++ | +++ |
| GK1.5 | +++ | ++ | |
| RM4-4 | nd | ||
| CD8 | 53-6.7 | +++ | ++ |
| CD11b | M1/70 | ++ | ++ |
| CD11c | N418 | +++ | +++ |
| CD19 | 1D3 | +++ | ++ |
| MB19-1 | ++ | - | |
| B220 | RA3-6B2 | +++ | ++ |
| GR-1 | RB6-8C5 | +++ | +++ |
| NK1.1 | PK136 | ++ | + |
| CD39 | 24DMS1 | + | - |
| CD49b | DX5 | ++ | - |
| CD25 | PC61.5 | + | - |
| CD69 | H1.2F3 | ++ | - |
| CXCR5 | SPRCL5 | ++ |
- |
| RAT | |||
| CD25 | OX39 | - | |
| CANINE | |||
| CD4 | ykix302.9 | +++ | + |
| CD5 | ykix322.3 | +++ | - |
| CD8a | ycate55.9 | +++ | ++ |
| CD44 | ykix337.8 | +++ | ++ |
| CD45 | ykix716.13 | +++ | ++ |
| CD45R | ykix753.22 | +++ | ++ |
| CD90 | ykix337.217 | +++ | ++ |
| MHC Class II | ykix334.2 | +++ | +++ |
| All testing carried out with eBioscience Fixation & Permeabilization Kit | |||
