固定破膜对细胞表面抗体结合的影响
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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). | |
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