Tag Antibodies

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HA Tag antibodies

[edit]
Accession Name Clone
Heavy Light Epitope
1FRG
12CA5
FAB 26/9

EVLLVESGGDLVKPGGFLKLSCAASGFTFSSFGMS
WVRHTPDKRLEWVATISNGGGYTYYQDSVKGRFT
ISRDNAKNTLFLEMTSLKSEDAGLYYCARRERYDE
KGFAYWGRGTLVTVSAAKTTAPSVYPLAPVCGDTT
GSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTF
PAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPA
SSTKVDKKIEPR

DIVMTQSPSSLTVTAGEKVTMSCKSSQSLFNSG
KRKNFLTWYHQKPGQPPKLLIYWASTRESGVPDRF
SGSGSGTDFTLT
ITSVQAEDLAIYYCQND
YSHPLTFGAGTKLELKRADAAPTVSIFPPSSEQL
TSGGASVVCFLNNFYPKDINVKWKIDGSERQNGV
LNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYT
CEATHKTSTSPIVKSFNR

XDVPDYASLX
1IFH

FAB 17/9
EVQLVESGGDLVKPGGSLKLSCAASGFSFSSYGM
SWVRQTPDKRLEWVATISNGGGYTYYPDSVKGR
FTISRDNAKNTLYLQMSSLKSEDSAMYYCARRE
RYDENGFAYWGQGTLVTVSAAKTTAPSVYPLAP
VCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSL
SSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNV
AHPASSTKVDKKIEPR

DIVMTQSPSSLTVTAGEKVTMSCTSSQSLFNSGKQK
NYLTWYQQKPGQPPKVLIYWASTRESGVPDRFTGS
GSGTDFTLTISSVQAEDLAVYYCQNDYSNPLTFGGG
TKLELKRADAAPTVSIFPPSSEQLTSGGASVVCFLNN
FYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYS
MSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRN

XDVPDYAS
Heavy

1FRG   1    EVLLVESGGDLVKPGGFLKLSCAASGFTFSSFGMSWVRHTPDKRLEWVATISNGGGYTYY  60
            EV LVESGGDLVKPGG LKLSCAASGF+FSS+GMSWVR TPDKRLEWVATISNGGGYTYY
1IFH   1    EVQLVESGGDLVKPGGSLKLSCAASGFSFSSYGMSWVRQTPDKRLEWVATISNGGGYTYY  60

1FRG   61   QDSVKGRFTISRDNAKNTLFLEMTSLKSEDAGLYYCARRERYDEKGFAYWGRGTLVTVSA  120
             DSVKGRFTISRDNAKNTL+L+M+SLKSED+ +YYCARRERYDE GFAYWG+GTLVTVSA
1IFH   61   PDSVKGRFTISRDNAKNTLYLQMSSLKSEDSAMYYCARRERYDENGFAYWGQGTLVTVSA  120

Light

1FRG   1    DIVMTQSPSSLTVTAGEKVTMSCKSSQSLFNSGKRKNFLTWYHQKPGQPPKLLIYWASTR  60
            DIVMTQSPSSLTVTAGEKVTMSC SSQSLFNSGK+KN+LTWY QKPGQPPK+LIYWASTR
1IFH   1    DIVMTQSPSSLTVTAGEKVTMSCTSSQSLFNSGKQKNYLTWYQQKPGQPPKVLIYWASTR  60

1FRG   61   ESGVPDRFSGSGSGTDFTLTITSVQAEDLAIYYCQNDYSHPLTFGAGTKLELKRADAAPT  120
            ESGVPDRF+GSGSGTDFTLTI+SVQAEDLA+YYCQNDYS+PLTFG GTKLELKRADAAPT
1IFH   61   ESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSNPLTFGGGTKLELKRADAAPT  120


Tag Antibodies

[edit]
Urgency Name Chain Clone ID Name Parent Vector Insert Size Enzymes Status/note
0 Vector
L 3154
pCMV5-HA-VH-mKappa
HA  3129 pMiniT2-3129-1
2493 pCMV5-mIgG-kLC
527 BamH1-Kpn1
sequenced, good, glycerol stock made
3
H 3129 
pCMV5-HA-VH-mIgG2b
new 3129 pMiniT with b 3154 pCMV5-Flag-H
1034
Kpn1 + Not1
3129-BamH1-F + 3129-Kpn1-F1
-2
L 3021 pCEL0-mIg-LCKappa

3016 pCEL0

Vector 3016 pCEL0:   BssHII  - Nhe1
insert in pMiniT2:  BssHII  - Xba1

PCR from  IgG kappa LC from 2493 pCMV5-mIgG-kLC, 3021-F2  + 3021-Age1-Xba1-R , Vector: BssHII - Nhe1,  insert: BssHII - Xba1, in  pMiniT2 vector now.
-2
H 3027 
pCEH0-mIgG2b-HC
3076-pMiniT2
3017 pCEH0

Nhe1-Not1

PCR from 2492  pCMV5-mIgG2b-HC  using 3020-Xba1-Nhe1-F + 3027-2b-Not1-Age1-R

in pMiniT2 now, sequenced

0 His H+L, sc 3002
pJet1.2-Anti-His Tag 3D5

pJet1.2

Nde1-Sal1/Xho1 3129-Kpn1-F1, sequenced, good, need to subclone into pET30b, but there is no tag for purification
0 His  L 3001 
pMiniT2-His-VL 




good, sequenced

0
H
3135
3135-3-pMiniT2

pMiniT2


in pMiniT2 vector now, 3135-3-pMiniT2 sequenced, good,  need to subclone into

3129 pCMV5-HA-VH-mIgG2b using BsrG1-Kpn1

0
L 3001 
pCMV5-His-VL-kappa
3001-pMiniT2
3130 pCMV5-HA-VL-mIgKappa
435 BamH1-Kpn1

3001-pMiniT2 is good, sequenced

subclone from 3001-pMiniT2, 

-2
H 3135 pCMV5-His-VH-IgG2b
 3135-3-pMiniT2 

3129 pCMV5-HA-VH-mIgG2b

3009pCMV5-Flag-VH-IgG2b

364 BsrG1-Kpn1 
subclone into 3129 pCMV5-HA-VH-mIgG2b using BsrG1-Kpn1 from 3135-3-pMiniT2 
0
H 3168
pCEH0-His-VH-mIgG2a
3135-3-pMiniT2 
3163 pCEH0-1FRG-HA-VH-mIgG2a
358 BsrG1-Nhe1

3135-3-pMiniT2 

VHSQVQLQQSGPEDVKPGASVKISCKASGYTFTDYYMNWVKQSPGKGLEWIGDINPNNGGTSYNQKFKGRATLTVDKSSSTAYMELRSLTSEDSSVYYCESQSGAYWGQGTTVTVSAASS

0 Myc
L 3003 
3003-L-5

pJet1.2


IL2 signal peptide, BamH1-Kpn1, good
0
H 3003 3003-H-1

pJet1.2


need to use 3076-myc-BsrG1-F to fix frame shift
0
L 3075
pCMV5-Myc-L-kappa

2493 pCMV5-mIgG-kLC


completed, subclone from 3003-L5 using BamH1-Kpn1 to 2493 pCMV5-mIgG-kLC

sequenced, good

-2
H 3076
pCMV5-Myc-H-IgG2b

3129 pCMV5-HA-VH-mIgG2b


 BsrG1-Kpn1 

PCR from 3003-H-1 in pJet1.2, using 3076-myc-BsrG1-F  and  3076-myc-Nhe1-Kpn1-R

in pMiniT2 now sequenced, good


0
H 3167
pCEH0-Myc-VH-mIgG2a

3027-pMiniT2

(3076 myc pMiniT2)

3163  pCEH0-1FRG-HA-VH-mIgG2a
397 BsrG1-Nhe1

subcloning from the misslabeled myc clone in pMiniT2

EVDLVESGGDLVKPGGSLKLSCAASGFTFSHYGMSWVRQTPDKRLEWVATIGSRGTYTHYPDSVKGRFTISRDNDKNALYLQMNSLKSEDTAMYYCARRSEFYYYGNTYYYSAMDYWGQGTTVTVSS

0 HA, 1IFH
sc VH+VL 3074 
pT7A30-HA-Avi-BirA

pJet1.2

3074-1-PJET1-2 is good, need to subclone into 1834 pT7A30-Avi-BirA
0 HA, 1IFH
L 3011
pJet1.2-HA-L-1

pJet1.2


3011-L-1 is good, sequenced
0 HA, 1IFH
H 3011
pJet1.2-HA-H-2

pJet1.2


3011-H-2 is good, sequenced

0 HA, 1IFH
L 3030 3130-4-pMiniT2 

pMiniT2


3130-4-pMiniT2 is good (sequenced), derived from 3011-L-1  pJet1.2-HA light chain by PCR using 3130-BamH1-R + 3130-Kpn1-R
0 HA, 1IFH
L 3130
pCMV5-HA-VL-mIgKappa
 3130-4-pMiniT2  
2493 pCMV5-mIgG-kLC
429bp
subclone  from using BamH1+ Kpn1

completed,good, sequenced, need to make glycerol stock

0 HA, 1IFH
H 3129 pMiniT2-3129-1 3011 pJet1.2-HA Heavy chain
pMiniT2

PCR good sequenced
0 HA, 1IFH
H 3129
pCMV5-HA-VH-mIgG2b
2492 pCMV5-mIgG2b-HC 3154 pCMV5-HA-VH-mKappa
  subcloning  Kpn1 -Not1  subcloning mIgG2b constant domain to 3154, due to BamH1 site
0 HA, 1IFH
L 3131 
pCEL0-HA-VL-mIgKappa
3011 pJet1.2-HA-L-1 
3021 pCEL0-mIg-LCKappa
342
BssHII-Xho1

subclone 342 bp fragment from 3011 pJet1.2-HA-L-1 using BssHII-Xho1

do not use the smallest fragment

0 HA, 1IFH
H 3132 
pCEH0-HA-VH-mIgG2a
3011 pJet1.2-HA-H-2
3163 pCEH0-1FRG-HA-VH-mIgG2a
373
BsrG1-Nhe1
subclone 373 bp fragment from 3011 pJet1.2-HA-H-2 using BsrG1-Nhe1
0 HA, 1FRG
L 3159
 pMiniT2-1FRG-HA-VL-kappa

pMiniT2

gene fragment from IDT
0 HA, 1FRG
H 3160 
pMiniT-2-1FRG-HA-VH-mIgG2a

pMiniT2


gene fragment from IDT
0 HA, 1FRG
L 3162
  pCMV5-1FRG-HA-VL-kappa

3130 pCMV5-HA-VL-mIgKappa
767 BamH1-Not1  subcloning from 3159
1 HA, 1FRG
H 3158
pCMV5-HA-VH-mIgG2a

3129 pCMV5-HA-VH-mIgG2b
1471 BsrG1-Not1 subcloning from 3160
0 HA, 1FRG
H 3163
pCEH0-1FRG-HA-VH-mIgG2a
3160 pMiniT-2-1FRG-HA-VH-mIgG2a
3017 pCEH0
1464 BsrG1-Not1
subcloning from 3160
0 HA, 1FRG
L 3164 pCEL0-1FRG-HA-VL-kappa
3159 pMiniT2-1FRG-HA-VL-kappa
 3016 pCEL0
701 BssHII --NheI
subcloning from 3159
0 Flag
L 3010  
3010-5-miniT2
pMiniT2

sequenced, good
0
H 3009
3009-4-pMiniT2-Flag-H 

pMiniT2


3009-4-pMiniT2, good, sequenced
0
L 3010 pCMV5-Flag-kappa
3010-5-miniT2

2493 pCMV5-Flag-kappa

3130

pCMV5-HA-VL-mIgKappa


BamH1+ Kpn1
need to do subcloning from 3010-5-miniT2
-2
H 3009 pCMV5-Flag-VH-IgG2b
3009-4-pMiniT2-Flag-H 


3129 pCMV5-HA-VH-mIgG2b

 432 BsrG1-Kpn1

subclone from 3009-4-pMiniT2 using BsrG1-Kpn1

0
H 3152
pCEH0-Flag-VH-mIgG2a
3009-4-pMiniT2 
3163 pCEH0-1FRG-HA-VH-mIgG2a
371

BsrG1-Nhe1

BsrG1-Kpn1

subclone from 3009-4-pMiniT2 using BsrG1-Nhe1

VHSEVQLQQSGGELAKPGASVKMSCKSSGYTFTAYAIHWAKQAAGAGLEWIGYIAPAAGAAAYNAAFKGKATLAADKSSSTAYMAAAALTSEDSAVYYCARAAAAGADYWGQGTTLTVSSAS

--Paul.xu (talk) 07:20, 13 February 2018 (EST)