有些时候，表面抗体需要在细胞固定之后标记，但这取决于固定过程对于要研究的蛋白是否会产生影响。
下表列出了eBioscience的几种固定过程，包括使用 Intracellular Fixation & Permeabilization Buffer Set (cat. no. 88-8824) 或者 IC Fixation Buffer (cat. no. 00-8222) 加甲醇处理，对于标记的影响状况。
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
|
++
|
++*
|
++
|
|
| 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).
|
|
