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Sometimes, surface antibodies need to be labeled after cell fixation, but this depends on whether the immobilization process has an effect on the protein to be studied. The following table lists several immobilization procedures for eBioscience, including the use of Intracellular Fixation & Permeabilization Buffer Set (cat. no. 88-8824) or IC Fixation Buffer (cat. no. 00-8222) plus methanol for labeling effects. situation. Antibody Clone Performance Following Fixation / Permeabilization |
Antigen | Clone | Live cells (Before IC Fixation)1 | After IC Fixation
And Perm Wash1 | After IC Fixation
/ Methanol2 | HUMAN | | | | | CD3 | OKT3 | +++ | ++ | + | | UCHT1 | +++ | +++ | +++ | | SK7 | +++ | ++* | +++ | CD4 | OKT4 | +++ | +* | +/- | | RPA-T4 | +++ | +++* | + | | SK3 | +++ | +++ | + | CD5 | UCHT2 | +++ | +++ | +++ | CD8 | OKT8 | +++ | +/- | - | | RPA-T8 | +++ | +++ | +/- | | SK1 | +++ | +++* | +/- | CD8b | SIDI8BEE | +++ | ++ | ++ | CD11a | HI111 | +++ | +++ | + | CD11b | CBRM1/5 | + | - | +++ | | ICRF44 | ++ | - | ++ | CD11c | 3.9 | ++ | +* | ++ | CD14 | 61D3 | +++ | +++* | ++ | CD19 | HIB19 | +++ | ++* | +/- |
CD20 | 2H7 | +++ | +++ | - | CD25 | BC96 | + | - | +/- | CD27 | O323 | +++ | +* | + | CD31 | WM59 | +++ | ++* | +++ | CD33 | P67.6 | +++ | +++* | +++ | CD38 | HB7 | ++ | - | ++ | | HIT2 | ++ | - | ++ | CD40 | 5C3 | ++ | - | ++ | CD44 | IM7 | +++ | +++ | +++ | CD45 | 2D1 | +++ | +++ | +++ | | HI30 | +++ | +++ | +++ | CD45RA | HI100 | +++ | +++ | +++ | CD45RO | UCHL1 | +++ | +++ | ++ | CD62L | DREG-56 | ++ | - | ++ | CD56 | MEM-188 | + | - | +/- | | CB56 | + | - | +/- | | CMSSB | ++ | +/-* | - | CD57 | TB01 | ++ | ++ | ++ | CD62L | DREG56 | ++ | - | | CD69 | FN50 | +++ | ++ | ++ | CD80 | 2D10.4 | + | - | + | CD83 | HB15e | ++ | ++ | ++ | CD86 | IT2.2 | + | +* | + | CD94 | HP-3D9 | ++ | ++ | ++ | CD95 | DX2 | + | - | + | CD127 | RDR5 | ++ | +* | ++ | CD161 | HP-3G10 | ++ | ++* | + |
MOUSE | | | | | CD3 | 145-2C11 | ++ | + | +/- | | 500A2 | +++ | +++* | +++ | | 17A2 | +++ | +++ | +++ | CD4 | GK1.5 | +++ | ++* | +++ | | RM4-5 | +++ | +++* | +++ | CD8 | 53-6.7 | +++ | ++ | ++ | CD11b | M1/70 | ++ | ++ | ++ | CD11c | N418 | +++ | +++ | ++ | CD19 | 1D3 | +++ | ++* | - | | MB19-1 | ++ | - | - | CD24 | M1/69 | +++ | +++ | ++ | CD25 | PC61.5 | + | - | + | | 3C7 | ++ | - | + | | 7D4 | ++ | ++ | + | CD39 | 24DMS1 | + | - | ++ | CD44 | IM7 | ++ | ++* | ++ | CD45 | 30-F11 | +++ | +++ | +++ | CD45R
(B220) | RA3-6B2 | +++ | ++ | +++ | CD49b | DX5 | ++ | - | + | CD69 | H1.2F3 | ++ | - | - | CD185
(CXCR5) | SPRCL5 | ++ | - | - | GR-1 | RB6-8C5 | +++ | +++ | ++ | NK1.1 | PK136 | ++ | + | + | IgD | 11-26c | +++ | +++* | ++ | IgM | Ll/41 | ++ | ++* | ++ | TCR
Beta | H57-597 | ++ | ++* | ++ |
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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 | +++ | +++ | | 1 Testing carried out with eBioscience Fixation & Permeabilization Kit for 15-30 minutes at room temperature followed by a Perm wash, see Best Protocols here (Protocol A). *Please note the clones with * indicate longer fixations are detrimental to staining. 2 Testing carried out with 15-30 minutes at room temperature in 50% IC Fixation followed by 30 minutes 90-100% Methanol incubation on ice, see Best Protocolshere (Protocol C). |
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