(A) Phospho-specific antibodies
|
Substrate
|
poY location
|
Mol mass (kDa)
|
Co.; Cat. #a
|
MM/RP/GPb
|
cortactin
|
421
|
80/85
|
BioS; 44–854
|
RP
|
paxillin
|
31
|
68
|
BioS; 44–720
|
RP
|
paxillin
|
118
|
68
|
BioS; 44–722
|
RP
|
Shc
|
239/240
|
46, 52, 66
|
BioS; 44–830
|
RP
|
PKC-δ
|
311
|
77
|
BioS; 44–950
|
RP
|
PLC-γ1
|
783
|
130
|
Bios; 44–696
|
RP
|
p56Dok-2
|
351
|
56, 58
|
CS; 3911
|
RP
|
CAS
|
410
|
130
|
CS; 4011
|
RP
|
connexin-43
|
265
|
43
|
SC; sc-17220
|
GP
|
p120catenin
|
228
|
120
|
BD; 612536
|
MM
|
vinculin
|
100
|
130
|
BioS; 44–1074G
|
RP
|
vinculin
|
1065
|
130
|
BioS; 44–1078G
|
RP
|
(B) Antibodies used for IP
|
Substrate
|
Mol mass (kDa)
|
Co.; Cat. #
a
|
MM/RP
b
| |
Sam68
|
68
|
SC; sc-333
|
RP
| |
Annexin II (H-50)
|
36(90)
|
SC; sc-9061
|
RP
| |
p190Rho-GAP
|
190
|
BD; 610149
|
MM
| |
Eps8
|
97
|
BD; 610143
|
MM
| |
Crk (Crk II)
|
40
|
BD; 610035
|
MM
| |
(C) Antibodies used for total substrate protein level
|
Substrate
|
Mol mass (kDa)
|
Co.; Cat. #
a
|
MM/RP
b
| |
cortactin
|
80/85
|
CS; 3502
|
RP
| |
paxillin
|
68
|
BD; 610551
|
MM
| |
Shc
|
46, 52, 66
|
CS; 2432
|
RP
| |
PKC-δ
|
77
|
SC; sc-213
|
RP
| |
PLC-γ1
|
130
|
CS; 2822
|
RP
| |
p56Dok-2
|
56, 58
|
CS; 3914
|
RP
| |
CAS
|
130
|
BD; 610271
|
MM
| |
p120catenin
|
120
|
BD; 610133
|
MM
| |
vinculin
|
130
|
Sigma; V-9131
|
MM
| |
GAPDH
|
37
|
SC; sc-25778
|
RP
| |
(D) Epitope-tag antibodies
|
Epitope tag
|
Co.; Cat. #
a
|
MM/RP
b
| | |
GST
|
SC; sc-138
|
MM
| | |
FLAG
|
Sigma; F3040
|
MM
| | |
GFP
|
Invitrogen; A6455
|
RP
| | |
-
a BioS – BioSource; CS – Cell Signaling; SC – Santa Cruz; BD – BD Biosciences
-
b Abs: RP – Rabbit polyclonal; MM – Mouse monoclonal; GP – Goat polyclonal
- * Abs to actinin (Sigma; A5044), CASpoY165 (CS; 4015), CASpoY249 (CS; 4014), p120cateninpoY96 (BD; 612534), p120cateninpoY280 (BD; 612538), ezrinpoY353 (CS; 3144) did not recognize appropriately-sized bands.