https://eln.excellgen.com/index.php?action=history&feed=atom&title=Mouse_Antibody_Cloning Mouse Antibody Cloning - Revision history 2026-10-18T04:24:16Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=Mouse_Antibody_Cloning&diff=17931&oldid=prev Paul.xu at 20:31, 12 May 2018 2018-05-12T20:31:54Z <p></p> <p><b>New page</b></p><div>== Mouse Light Chain ==<br /> <br /> == Alignment ==<br /> &lt;pre&gt;K --ADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQ 58<br /> L1 GQPKSSPSVTLFPPSSEELETNKATLVCTITDFYPGVVTVDWKVDGTPVTQGMETTQPSK 60<br /> L2 GQPKSTPTLTVFPPSSEELKENKATLVCLISNFSPSGVTVAWKANGTPITQGVDTSNPTK 60<br /> .::*::::******:* . *::** :.:* * :.* ** :*: ;:*: .: :<br /> <br /> K DSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC- 106<br /> L1 Q-SNNKYMASSYLTLTARAWERHSSYSCQVTHEGHT--VEKSLSRADCS 106<br /> L2 E--GNKFMASSFLHLTSDQWRSHNSFTCQVTHEGDT--VEKSLSPAECL 105<br /> : ...: ** * ** :. *.*::*:.**: * : **:. :* <br /> <br /> L1 GQPKSSPSVTLFPPSSEELETNKATLVCTITDFYPGVVTVDWKVDGTPVTQGMETTQPSK 60<br /> L2 GQPKSTPTLTVFPPSSEELKENKATLVCLISNFSPSGVTVAWKANGTPITQGVDTSNPTK 60<br /> *****:*::*:********: ******* *::* *. *** **.:***:***::*::*:*<br /> <br /> L1 QSNNKYMASSYLTLTARAWERHSSYSCQVTHEGHTVEKSLSRADCS 106<br /> L2 E-GNKFMASSFLHLTSDQWRSHNSFTCQVTHEGDTVEKSLSPAECL 105<br /> : .**:****:* **: *. *.*::*******.******* *:* <br /> &lt;/pre&gt;<br /> <br /> == ==<br /> <br /> == Oligo for RT: Light chain (kappa &amp; lambda) ==<br /> &lt;pre&gt;t(g/c)ctcactggatggtgggaa<br /> tSctcactggatggtgggaa<br /> {ttcccaccatccagtgag (c/g)a}<br /> F P P S S E Q<br /> &lt;/pre&gt;<br /> <br /> == Kappa ==<br /> <br /> constant region<br /> <br /> GCAGATGCTGCACCAACTGTATCCATCTTCCCACCATCCAGTGAGCAGTTAACATCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTTCTACCCCAAAGACATCAATGTCAAGTGGAAGATTGATGGCAGTGAACGACAAAATGGCGTCCTGAACAGTTGGACTGATCAGGACAGCAAAGACAGCACCTACAGCATGAGCAGCACCCTCACGTTGACCAAGGACGAGTATGAACGACATAACAGCTATACCTGTGAGGCCACTCACAAGACATCAACTTCACCCATTGTCAAGAGCTTCAACAGGAATGAGTGTTAG<br /> <br /> ADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDS KDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC<br /> &lt;pre style=&quot;color: rgb(98, 98, 98); background-color: rgb(255, 255, 255);&quot;&gt;gcagatgctgcaccaactgtatccatcttcccaccatccagtgagcagttaacatctgga<br /> A D A A P T V S I F P P S S E Q L T S G <br /> ggtgcctcagtcgtgtgcttcttgaacaacttctaccccaaagacatcaatgtcaagtgg<br /> G A S V V C F L N N F Y P K D I N V K W <br /> aagattgatggcagtgaacgacaaaatggcgtcctgaacagttggactgatcaggacagc<br /> K I D G S E R Q N G V L N S W T D Q D S <br /> aaagacagcacctacagcatgagcagcaccctcacgttgaccaaggacgagtatgaacga<br /> K D S T Y S M S S T L T L T K D E Y E R <br /> cataacagctatacctgtgaggccactcacaagacatcaacttcacccattgtcaagagc<br /> H N S Y T C E A T H K T S T S P I V K S <br /> ttcaacaggaatgagtgttag<br /> F N R N E C - &lt;/pre&gt;<br /> <br /> === ===<br /> &lt;pre&gt;aligment of some kappa light chain VL domain<br /> 4 elvltqspttmaaspgekititcsasssissnylhwyqqkpgfspklliyrtsnlasgvp<br /> 2 qivltqspaimsaslgdrvtmtctasssvsssylhwyqqkpgsspklwiystsnlasgvp<br /> 1 envltqspaimsaspgekvtmacrasssvsstylhwyqqksgaspklliystsnlasgvp<br /> 3 envltqspaimsaspgekvtmtcrasssvsssylhwyqqksgaspklwiystsnlasgvp<br /> 5 -ivmtqtqkfmstsvgervsitckasqnvgt-nvawyqqkggqslelliygasnrhtgvp<br /> 6 dvvmtqthkfmstsvgdrvsitckasqdvsg-avawyqqksgqspkllismasqrytgvp<br /> *:**: *::* *::::::* **..:. : ***** * * :* * :*: :***<br /> <br /> 4 arfsgsgsgtsysltigtmeaedvatyycqqgssipftfgsgtkleikr<br /> 2 arfsgsgsgtsysltissmeaedaatyychqfhr-sltfgsgtkleikr<br /> 1 arfsgsgsgtsysltissveaedaatyycqqysgypltfgagtklelkr<br /> 3 arfsgsgsgtsysltissveaedaatyycqqysgypytfgggtkleikr<br /> 5 drftgrgsgtdftltitnvqsedmtnyfceqysrypltfgagtklelkr<br /> 6 drftgsgsgtdftftissvqaedlavyycqqhyaipltfgagtklelkr<br /> **:* ****.:::** .:::** : *:*.* ***.*****:**<br /> <br /> &lt;/pre&gt;<br /> <br /> === Lambda 1 ===<br /> <br /> GGCCAGCCCAAGTCTTCGCCATCAGTCACCCTGTTTCCACCTTCCTCTGAAGAGCTCGAGACTAACAAGGCCACACTGGTGTGTACGATCACTGATTTCTACCCAGGTGTGGTGACAGTGGACTGGAAGGTAGATGGTACCCCTGTCACTCAGGGTATGGAGACAACCCAGCCTTCCAAACAGAGCAACAACAAGTACATGGCTAGCAGCTACCTGACCCTGACAGCAAGAGCATGGGAAAGGCATAGCAGTTACAGCTGCCAGGTCACTCATGAAGGTCACACTGTGGAGAAGAGTTTGTCCCGTGCTGACTGTTCCTAG<br /> <br /> GQPKSSPSVTLFPPSSEELETNKATLVCTITDFYPGVVTVDWKVDGTPVTQGMETTQPSKQSNNKYMASSYLTLTARAWERHSSYSCQVTHEGHTVEKSLSRADCS<br /> &lt;pre style=&quot;color: rgb(98, 98, 98); background-color: rgb(255, 255, 255);&quot;&gt;ggccagcccaagtcttcgccatcagtcaccctgtttccaccttcctctgaagagctcgag<br /> G Q P K S S P S V T L F P P S S E E L E <br /> actaacaaggccacactggtgtgtacgatcactgatttctacccaggtgtggtgacagtg<br /> T N K A T L V C T I T D F Y P G V V T V <br /> gactggaaggtagatggtacccctgtcactcagggtatggagacaacccagccttccaaa<br /> D W K V D G T P V T Q G M E T T Q P S K <br /> cagagcaacaacaagtacatggctagcagctacctgaccctgacagcaagagcatgggaa<br /> Q S N N K Y M A S S Y L T L T A R A W E <br /> aggcatagcagttacagctgccaggtcactcatgaaggtcacactgtggagaagagtttg<br /> R H S S Y S C Q V T H E G H T V E K S L <br /> tcccgtgctgactgttcctag<br /> S R A D C S -&lt;/pre&gt;<br /> <br /> === Lambda 2 ===<br /> <br /> GGTCAGCCCAAGTCCACTCCCACTCTCACCGTGTTTCCACCTTCCTCTGAGGAGCTCAAGGAAAACAAAGCCACACTGGTGTGTCTGATTTCCAACTTTTCCCCGAGTGGTGTGACAGTGGCCTGGAAGGCAAATGGTACACCTATCACCCAGGGTGTGGACACTTCAAATCCCACCAAAGAGGGCAACAAGTTCATGGCCAGCAGCTTCCTACATTTGACATCGGACCAGTGGAGATCTCACAACAGTTTTACCTGCCAAGTTACACATGAAGGGGACACTGTGGAGAAGAGTCTGTCTCCTGCAGAATGTCTCTAA <br /> <br /> GQPKSTPTLTVFPPSSEELKENKATLVCLISNFSPSGVTVAWKANGTPITQGVDTSNPTKEGNKFMASSFLHLTSDQWRSHNSFTCQVTHEGDTVEKSLSPAECL<br /> <br /> <br /> &lt;pre style=&quot;color: rgb(98, 98, 98); background-color: rgb(255, 255, 255);&quot;&gt;ggtcagcccaagtccactcccactctcaccgtgtttccaccttcctctgaggagctcaag<br /> G Q P K S T P T L T V F P P S S E E L K <br /> gaaaacaaagccacactggtgtgtctgatttccaacttttccccgagtggtgtgacagtg<br /> E N K A T L V C L I S N F S P S G V T V <br /> gcctggaaggcaaatggtacacctatcacccagggtgtggacacttcaaatcccaccaaa<br /> A W K A N G T P I T Q G V D T S N P T K <br /> gagggcaacaagttcatggccagcagcttcctacatttgacatcggaccagtggagatct<br /> E G N K F M A S S F L H L T S D Q W R S <br /> cacaacagttttacctgccaagttacacatgaaggggacactgtggagaagagtctgtct<br /> H N S F T C Q V T H E G D T V E K S L S <br /> cctgcagaatgtctctaa<br /> P A E C L - &lt;/pre&gt;<br /> <br /> Alignment<br /> &lt;pre&gt;Lambda1 GQPKSSPSVTLFPPSSEELETNKATLVCTITDFYPGVVTVDWKVDGTPVTQGMETTQPSK<br /> lambda2 GQPKSTPTLTVFPPSSEELKENKATLVCLISNFSPSGVTVAWKANGTPITQGVDTSNPTK<br /> *****:*::*:********: ******* *::* *. *** **.:***:***::*::*:*<br /> <br /> Lambda1 QSNNKYMASSYLTLTARAWERHSSYSCQVTHEGHTVEKSLSRADCS<br /> lambda2 E-GNKFMASSFLHLTSDQWRSHNSFTCQVTHEGDTVEKSLSPAECL<br /> : .**:****:* **: *. *.*::*******.******* *:* &lt;/pre&gt;<br /> <br /> == Heavy Chain constant region ==<br /> &lt;pre&gt;gccaaaacaacagccccatcggtctatccactggcccctgtgtgtggagatacaactggc<br /> A K T T A P S V Y P L A P V C G D T T G <br /> tcctcggtgactctaggatgcctggtcaagggttatttccctgagccagtgaccttgacc<br /> S S V T L G C L V K G Y F P E P V T L T <br /> tggaactctggatccctgtccagtggtgtgcacaccttcccagctgtcctgcagtctgac<br /> W N S G S L S S G V H T F P A V L Q S D <br /> ctctacaccctcagcagctcagtgactgtaacctcgagcacctggcccagccagtccatc<br /> L Y T L S S S V T V T S S T W P S Q S I <br /> acctgcaatgtggcccacccggcaagcagcaccaaggtggacaagaaaatt<br /> T C N V A H P A S S T K V D K K I <br /> <br /> ctcagggaaataacccttgaccaggcatcctagagtcac<br /> (gtgactctaggatgcctggtcaagggttatttccctgag) <br /> V T L G C L V K G Y F P E<br /> <br /> Primer for RT<br /> Mouse-κγLC-RT tSctcactggatggtgggaa<br /> Mouse-IgG-HC-RT ctcagggaaataacccttgac<br /> CTC AGG GAA ATA ACC CTT GAC <br /> &lt;/pre&gt;<br /> <br /> == Alignment ==<br /> &lt;pre&gt;G1 AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSD 60<br /> G2a AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSD 60<br /> G2b AKTTPPSVYPLAPGCGDTTGSSVTLGCLVKGYFPESVTVTWNSGSLSSSVHTFPALLQSG 60<br /> **** ******** .. *.* ************* **:*********.******:***.<br /> <br /> G1 LYTLSSSVTVPSSPRPSETVTCNVAHPASSTKVDKKIVPRDC-----GCKPC----ICTV 111<br /> G2a LYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGP--TIKPCPPC----KCPA 114<br /> G2b LYTMSSSVTVPSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPCKECHKCPA 120<br /> ***:****** ** **:::**.********.****: * . * ** * .<br /> <br /> G1 P---EVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQP 168<br /> G2a PNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQT 174<br /> G2b PNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPDVRISWFVNNVEVHTAQTQT 180<br /> * *******: **** *:*:* *******:*:***:*::****::********* <br /> <br /> G1 REEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTI 228<br /> G2a HREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVL 234<br /> G2b HREDYNSTIRVVSALPIQHQDWMSGKEFKCKVNNKDLPSPIERTISKIKGLVRAPQVYIL 240<br /> :.*::***:* ** *** ****:.******:**. :*:***:**** ** :***** :<br /> <br /> G1 PPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMNTNGSYFVYSKLN 288<br /> G2a PPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLR 294<br /> G2b PPPAEQLSRKDVSLTCLVVGFNPGDISVEWTSNGHTEENYKDTAPVLDSDGSYFIYSKLD 300<br /> *** *::::..*:***::..* * ** *** **: ***:* *:::::****:**** <br /> <br /> G1 VQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK 324<br /> G2a VEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK 330<br /> G2b IKTSKWEKTDSFSCNVRHEGLKNYYLKKTISRSPGK 336<br /> ::..:* ::::*.* ****:*:: *::*::***<br /> &lt;/pre&gt;<br /> <br /> <br /> <br /> === IgG1 ===<br /> <br /> GCTAAAACGACACCCCCATCTGTCTATCCACTGGCCCCTGGATCTGCTGCCCAAACTAACTCCATGGTGACCCTGGGATGCCTGGTCAAGGGCTATTTCCCTGAGCCAGTGACAGTGACCTGGAACTCTGGATCCCTGTCCAGCGGTGTGCACACCTTCCCAGCTGTCCTGCAGTCTGACCTCTACACTCTGAGCAGCTCAGTGACTGTCCCCTCCAGCACCTGGCCCAGCGAGACCGTCACCTGCAACGTTGCCCACCCGGCCAGCAGCACCAAGGTGGACAAGAAAATTGTGCCCAGGGATTGTGGTTGTAAGCCTTGCATATGTACAGTCCCAGAAGTATCATCTGTCTTCATCTTCCCCCCAAAGCCCAAGGATGTGCTCACCATTACTCTGACTCCTAAGGTCACGTGTGTTGTGGTAGACATCAGCAAGGATGATCCCGAGGTCCAGTTCAGCTGGTTTGTAGATGATGTGGAGGTGCACACAGCTCAGACGCAACCCCGGGAGGAGCAGTTCAACAGCACTTTCCGCTCAGTCAGTGAACTTCCCATCATGCACCAGGACTGGCTCAATGGCAAGGAGTTCAAATGCAGGGTCAACAGTGCAGCTTTCCCTGCCCCCATCGAGAAAACCATCTCCAAAACCAAAGGCAGACCGAAGGCTCCGCAGGTGTACACCATTCCACCTCCCAAGGAGCAGATGGCCAAGGATAAAGTCAGTCTGACCTGCATGATAACAGACTTCTTCCCTGAAGACATTACTGTGGAGTGGCAGTGGAATGGGCAGCCAGCGGAGAACTACAAGAACACTCAGCCCATCATGGACACAGATGGCTCTTACTTCGTCTACAGCAAGCTCAATGTGCAGAAGAGCAACTGGGAGGCAGGAAATACTTTCACCTGCTCTGTGTTACATGAGGGCCTGCACAACCACCATACTGAGAAGAGCCTCTCCCACTCTCCTGGTAAATGA<br /> <br /> AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSPRPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMNTNGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK<br /> &lt;pre&gt;alignment of some IgG1 VH domain<br /> <br /> 6 -----------------------------qvqlkqsgpglvapsqslsitctvsgfslt-<br /> 7 matgsrtslllafgl---lclpwlqegsaqvtlkesgpgilrpsqtldltcsfsgfslst<br /> 2 ----------mnfglsliflvlilkgvqcevklvesggglvkpggslklscaasgftfs-<br /> 1 -----------------------------qvqlqqsgaelvrpgssvkisckasgyafs-<br /> 5 matgsrtslllafgl---lclpwlqegsaqvqlqqsgaelvkpgssvkisckasgytft-<br /> 3 ----------mgwswiflfllsgtagvlsevqlqqsgpervkpgasvkisckasgytft-<br /> 4 ----------mgwswiflfllsgtagvlsevqlqqsgpervkpgasvkisckasgytft-<br /> 8 ----------mgwtwifililsvttgvhsdvqlqqsgpelekpgasvkisckasgfslp-<br /> :* * :** *. ::.::* **::: <br /> <br /> 6 -gygvnwvrqspgkglewlgmiwg-dgrtdyksalksrlsitkdnsksqvflkmnslqtd<br /> 7 sglsvgwirqpsgkglewlahiww-ddvkyfnpslksrltiskdssrnqvflkitsvdta<br /> 2 -sygmswvrqtpekrlewvatisgggshtyypdivkgrftisrdnaknnlylqmsslrse<br /> 1 -sywmnwvkqrpgqglewigqiypgdsdtnyngkfkgkatltadkssstaymqlssltse<br /> 5 -sddmhwikqqpgnglewigwiypgnddtkynqkfngkatltadkssstvymqlssltse<br /> 3 -dyymnwvkqshgkslewigdinpnnggtsynqkfegkatltvdkssstaymelrsltse<br /> 4 -dyymnwvkqshgkslewigdinpnnggtsynqkfegkatltvdkssstaymelrsltse<br /> 8 -ghninwivqrngkslewignidpyyggtnfnpkfkgkatltvdkssstlymhltslqse<br /> . : *: * : ***:. * . . : .:.: ::: *.: . ::.: *: : <br /> <br /> 6 dtaryfcasdyyg-sgsfaywgqgtlvtvsa<br /> 7 dsatyhctrgplg-hg-fdywgqgtlvtvsa<br /> 2 dtalyhcarrdwd-gyyfdywgqgttltvss<br /> 1 dsavyfcardyg--kdhfdywgqgttltvss<br /> 5 dsavyfcargdlnyggsmdawgqgtsvtvss<br /> 3 dsavyycarg--g-lhwfpywgqgtlvtvsa<br /> 4 dsavyycarg--g-lhwfpywgqgtlvtvsa<br /> 8 dsavyycarrrdg-nygftywgqgtlvtvsa<br /> *:* *.*: : ***** :***:<br /> <br /> Cloning site:<br /> Nhe1<br /> GCTAGCAGC GCTAAAACGAC<br /> A S S A K T <br /> <br /> &lt;/pre&gt;<br /> <br /> === IgG2a ===<br /> <br /> GCTAAAACAACAGCCCCATCGGTCTATCCACTGGCCCCTGTGTGTGGAGATACAACTGGCTCCTCGGTGACTCTAGGATGCCTGGTCAAGGGTTATTTCCCTGAGCCAGTGACCTTGACCTGGAACTCTGGATCCCTGTCCAGTGGTGTGCACACCTTCCCAGCTGTCCTGCAGTCTGACCTCTACACCCTCAGCAGCTCAGTGACTGTAACCTCGAGCACCTGGCCCAGCCAGTCCATCACCTGCAATGTGGCCCACCCGGCAAGCAGCACCAAGGTGGACAAGAAAATTGAGCCCAGAGGGCCCACAATCAAGCCCTGTCCTCCATGCAAATGCCCAGCACCTAACCTCTTGGGTGGACCATCCGTCTTCATCTTCCCTCCAAAGATCAAGGATGTACTCATGATCTCCCTGAGCCCCATAGTCACATGTGTGGTGGTGGATGTGAGCGAGGATGACCCAGATGTCCAGATCAGCTGGTTTGTGAACAACGTGGAAGTACACACAGCTCAGACACAAACCCATAGAGAGGATTACAACAGTACTCTCCGGGTGGTCAGTGCCCTCCCCATCCAGCACCAGGACTGGATGAGTGGCAAGGAGTTCAAATGCAAGGTCAACAACAAAGACCTCCCAGCGCCCATCGAGAGAACCATCTCAAAACCCAAAGGGTCAGTAAGAGCTCCACAGGTATATGTCTTGCCTCCACCAGAAGAAGAGATGACTAAGAAACAGGTCACTCTGACCTGCATGGTCACAGACTTCATGCCTGAAGACATTTACGTGGAGTGGACCAACAACGGGAAAACAGAGCTAAACTACAAGAACACTGAACCAGTCCTGGACTCTGATGGTTCTTACTTCATGTACAGCAAGCTGAGAGTGGAAAAGAAGAACTGGGTGGAAAGAAATAGCTACTCCTGTTCAGTGGTCCACGAGGGTCTGCACAATCACCACACGACTAAGAGCTTCTCCCGGACTCCGGGTAAATGA<br /> <br /> AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK<br /> <br /> === IgG2b ===<br /> <br /> GCTAAAACAACACCCCCATCAGTCTATCCACTGGCCCCTGGGTGTGGAGATACAACTGGTTCCTCTGTGACTCTGGGATGCCTGGTCAAGGGCTACTTCCCTGAGTCAGTGACTGTGACTTGGAACTCTGGATCCCTGTCCAGCAGTGTGCACACCTTCCCAGCTCTCCTGCAGTCTGGACTCTACACTATGAGCAGCTCAGTGACTGTCCCCTCCAGCACCTGGCCAAGTCAGACCGTCACCTGCAGCGTTGCTCACCCAGCCAGCAGCACCACGGTGGACAAAAAACTTGAGCCCAGCGGGCCCATTTCAACAATCAACCCCTGTCCTCCATGCAAGGAGTGTCACAAATGCCCAGCTCCTAACCTCGAGGGTGGACCATCCGTCTTCATCTTCCCTCCAAATATCAAGGATGTACTCATGATCTCCCTGACACCCAAGGTCACGTGTGTGGTGGTGGATGTGAGCGAGGATGACCCAGACGTCCGGATCAGCTGGTTTGTGAACAACGTGGAAGTACACACAGCTCAGACACAAACCCATAGAGAGGATTACAACAGTACTATCCGGGTGGTCAGTGCCCTCCCCATCCAGCACCAGGACTGGATGAGTGGCAAGGAGTTCAAATGCAAGGTCAACAACAAAGACCTCCCATCACCCATCGAGAGAACCATCTCAAAAATTAAAGGGCTAGTCAGAGCTCCACAAGTATACATCTTGCCGCCACCAGCAGAGCAGTTGTCCAGGAAAGATGTCAGTCTCACTTGCCTGGTCGTGGGCTTCAACCCTGGAGACATCAGTGTGGAGTGGACCAGCAATGGGCATACTGAGGAGAACTACAAGGACACCGCACCAGTCCTGGACTCTGACGGTTCTTACTTCATATACAGCAAGCTCGATATAAAAACAAGCAAGTGGGAGAAAACAGATTCCTTCTCATGCAACGTGAGACACGAGGGTCTGAAAAATTACTACCTGAAGAAGACCATCTCCCGGTCTCCGGGTAAATGA<br /> <br /> AKTTPPSVYPLAPGCGDTTGSSVTLGCLVKGYFPESVTVTWNSGSLSSSVHTFPALLQSGLYTMSSSVTVPSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPCKECHKCPAPNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPDVRISWFVNNVEVHTAQTQTHREDYNSTIRVVSALPIQHQDWMSGKEFKCKVNNKDLPSPIERTISKIKGLVRAPQVYILPPPAEQLSRKDVSLTCLVVGFNPGDISVEWTSNGHTEENYKDTAPVLDSDGSYFIYSKLDIKTSKWEKTDSFSCNVRHEGLKNYYLKKTISRSPGK<br /> &lt;pre&gt;IgG1 AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSD<br /> IgG2a AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSD<br /> IgG2b AKTTPPSVYPLAPGCGDTTGSSVTLGCLVKGYFPESVTVTWNSGSLSSSVHTFPALLQSG<br /> **** ******** .. *.* ************* **:*********.******:***.<br /> <br /> IgG1 LYTLSSSVTVPSSPRPSETVTCNVAHPASSTKVDKKIVPRDC-----GCKPC----ICTV<br /> IgG2a LYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGP--TIKPCPPC----KCPA<br /> IgG2b LYTMSSSVTVPSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPCKECHKCPA<br /> ***:****** ** **:::**.********.****: * . * ** * .<br /> <br /> IgG1 P---EVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQP<br /> IgG2a PNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQT<br /> IgG2b PNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPDVRISWFVNNVEVHTAQTQT<br /> * *******: **** *:*:* *******:*:***:*::****::********* <br /> <br /> IgG1 REEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTI<br /> IgG2a HREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVL<br /> IgG2b HREDYNSTIRVVSALPIQHQDWMSGKEFKCKVNNKDLPSPIERTISKIKGLVRAPQVYIL<br /> :.*::***:* ** *** ****:.******:**. :*:***:**** ** :***** :<br /> <br /> IgG1 PPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIMNTNGSYFVYSKLN<br /> IgG2a PPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLR<br /> IgG2b PPPAEQLSRKDVSLTCLVVGFNPGDISVEWTSNGHTEENYKDTAPVLDSDGSYFIYSKLD<br /> *** *::::..*:***::..* * ** *** **: ***:* *:::::****:**** <br /> <br /> IgG1 VQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK<br /> IgG2a VEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK<br /> IgG2b IKTSKWEKTDSFSCNVRHEGLKNYYLKKTISRSPGK<br /> ::..:* ::::*.* ****:*:: *::*::***<br /> &lt;/pre&gt;<br /> <br /> === IgG2a VH domain alignment ===<br /> &lt;pre&gt;CBI70317 mtslfslqllstqdlamgwsciilflvaaagrvhsqvqlqqpgaelvkpgasvkmsckas<br /> ADA84145 ----------------mavlvlflclvafpscvlsqvqlkesgpglvapsqslsitctvs<br /> AGH20709 --------------------------mdwtwrvlcqitlkesgpgilqpsqtlsltcsfs<br /> CAD18854 ----------------mgrltssflllivpayvlsqvtlkesgpgilqpsqtlsltcsfs<br /> AAI08376 ----------------mylglscvfivfllkgvqcevkldesggglvqpgrsmklscvas<br /> <br /> <br /> CBI70317 gysftsywm--nwvkqrpgrglewigridpsd-n-ethynqdfkdkvtltvdkssstvyi<br /> ADA84145 gfsltnygv--hwvrqppgkglewlgmiwagg---stnynsalmsrlninkdnsksqvfl<br /> AGH20709 gfslttygigagwirqppgkglewlthiwwnd---nkyyntalrsrltiskdssnnqvll<br /> CAD18854 gfslstygmgvgwirqpsgkglewlaniwwnd---hkyynsalksrltiskdtsnnqvfl<br /> AAI08376 gftfsdywm--nwvhqspekglewvaqirnkpynyvtyysdsvrgrftisrddskssvyl<br /> <br /> <br /> CBI70317 qlssltsedsavyycgr----lgyv-ygfdywgqgttltvssakttaps<br /> ADA84145 emnslqtddtalyfcareat-sgyvdyavdywgqgtsvtvssakttaps<br /> AGH20709 kianvdtadtatyycl----------ygytywgqgtlvtvsaakttaps<br /> CAD18854 kissvdtadtatyycah----ysnyayaldywgqgtsvtvssakttaps<br /> AAI08376 qmnnlraedtgiyyctssntyysyedy-fdywgqgttitvssakttaps<br /> <br /> &lt;/pre&gt;<br /> <br /> === IgG2b VH domain alignment ===<br /> &lt;pre&gt;7 mdrltssflllivpayvlsqvtlkesgpgilkpsqtlsltcsfsgfslstsgmgvgwirq<br /> 1 mdwlwnllflmaaaqsaqaqiqlaqsgpelkepgetvkiscktsgftfityg--vswvkq<br /> 5 mkcswvifflmavvigvnsevqllqsgtelvrpgasvklsctasgfsikdsl--lhwvmq<br /> 4 mewpcillfllsvtegvhsqvqllqsgpelvkpgasvkiscrasgyafsksw--mnwvkr<br /> 9 mewiwiflfilsgtagvqsqvqlqqsgaelarpgasvklsckasgytltsyg--iswvkq<br /> 10 mewiwiflfilsgtagvryqvqlqqsgaelarpgasvklsckasgytltsyg--iswvkq<br /> 3 mgwscimlflaatatgvhsqvqlqqpgaelvmpgasvrlsckasgytftryw--mhwlkq<br /> 8 mewswiflfllsgtagvhsevqlqqsgpelvnpgasvkmsckasgytfityv--mhwvkq<br /> 2 merhwiflfllsvtagvhsqvqlqqsgaelakpgasvkmsckasgytftsyw--mhwvkq<br /> 6 merhwiflflfsvtagvhsqvqlqqsgaelakpgasvkmsckasgysftsyw--mhwvkq<br /> * ::: . :: * : * : *. :: ::* **::: : *: :<br /> <br /> 7 psgkglewlahiww-dddkhynpslksqltiskdssrnqvflkitsvdtadtatyycvrr<br /> 1 apgkglkwmgwintysglptyapdfkgrfafslktsastaylqinnlknedtatyfcvin<br /> 5 rpeqglewigwidpedgetkyvpkfqdkatltaatssntaylqlssltsgdtatyyclfy<br /> 4 rpgkglewigrifpgdgdthysgkfqgkakltadkssvtaflqltsltsedsavyfcard<br /> 9 rtgqglewigeiypgsgnsyfnekfkgkatltvdkssstaylhlssltsedsavyfcagp<br /> 10 rtgqglewigeiypgsgnsyfnekfkgkatltvdkssstaylhlssltsedsavyfcagp<br /> 3 rpgrglewigwidpnsdgtkynekfkgkatltvdipsstafmqlssltsedsavyycagn<br /> 8 kpgqglewigyinpnkdgtkfnekfkgkatltsdkssntaymelssltsedsavyycard<br /> 2 rpgqglewigyinpssgyteynqkfkdkatltadkssstaymqlssltsedsavyycars<br /> 6 rpgqglewigyinpstgyteynqkfkdkatltadkssstaymqlssltsedsavyycarg<br /> :**:*:. * . : .::.: :: .::.:..: . *:*.*:* <br /> <br /> 7 sfsygssrdyfdywgqgttltvssakttpp<br /> 1 dpy------amdywgqgtsvtvssakttpp<br /> 5 gydd--grdyfdywgqgttitvssakttpp<br /> 4 sdy----gdyfddwgqgatvtvssakttpp<br /> 9 rqv---gllpfgywgqgtlvtasaakttpp<br /> 10 rqv---gllpfgywgqgtlvtasaakttpp<br /> 3 fg---------aywgqgtlvtvsaakttpp<br /> 8 ydy-----dwfaywgqgtlvtvsaakttpp<br /> 2 fyyy--segamdywgqgtsvtvssakttpp<br /> 6 dgyy---egwfaywgqgtlvtvsaakttpp<br /> ****: :*.*:******<br /> &lt;/pre&gt;<br /> <br /> == Mouse IgM ==<br /> <br /> SQSFPNVFPLVSCESPLSDKNLVAMGCLARDFLPSTISFTWNYQNNTEVIQGIRTFPTLRTGGKYLATSQVLLSPKSILEGSDEYLVCKIHYGGKNRDLHVPIPAVAEMNPNVNVFVPPRDGFSGPAPRKSKLICEATNFTPKPITVSWLKDGKLVESGFTTDPVTIENKGSTPQTYKVISTLTISEIDWLNLNVYTCRVDHRGLTFLKNVSSTCAASPSTDILTFTIPPSFADIFLSKSANLTCLVSNLATYETLNISWASQSGEPLETKIKIMESHPNGTFSAKGVASVCVEDWNNRKEFVCTVTHRDLPSPQKKFISKPNEVHKHPPAVYLLPPAREQLNLRESATVTCLVKGFSPADISVQWLQRGQLLPQEKYVTSAPMPEPGAPGFYFTHSILTVTEEEWNSGETYTCVVGHEALPHLV<br /> <br /> secreted form TERTVDKSTGKPTLYNVSLIMSDTGGTCY<br /> <br /> membrane bound form EGEVNAEEEGFENLWTTASTFIVLFLLSLFYSTTVTLFKVK<br /> <br /> Secreted Form from BALB/c AGAGTCAGTCCTTCCCAAATGTCTTCCCCCTCGTCTCCTGCGAGAGCCCCCTGTCTGATAAGAATCTGGTGGCCATGGGCTGCCTAGCCCGGGACTTCCTGCCCAGCACCATTTCCTTCACCTGGAACTACCAGAACAACACTGAAGTCATCCAGGGTATCAGAACCTTCCCAACACTGAGGACAGGGGGCAAGTACCTAGCCACCTCGCAGGTGTTGCTGTCTCCCAAGAGCATCCTTGAAGGTTCAGATGAATACCTGGTATGCAAAATCCACTACGGAGGCAAAAACAGAGATCTGCATGTGCCCATTCCAGCTGTCGCAGAGATGAACCCCAATGTAAATGTGTTCGTCCCACCACGGGATGGCTTCTCTGGCCCTGCACCACGCAAGTCTAAACTCATCTGCGAGGCCACGAACTTCACTCCAAAACCGATCACAGTATCCTGGCTAAAGGATGGGAAGCTCGTGGAATCTGGCTTCACCACAGATCCGGTGACCATCGAGAACAAAGGATCCACACCCCAAACCTACAAGGTCATAAGCACACTTACCATCTCTGAAATCGACTGGCTGAACCTGAATGTGTACACCTGCCGTGTGGATCACAGGGGTCTCACCTTCTTGAAGAACGTGTCCTCCACATGTGCTGCCAGTCCCTCCACAGACATCCTAACCTTCACCATCCCCCCCTCCTTTGCCGACATCTTCCTCAGCAAGTCCGCTAACCTGACCTGTCTGGTCTCAAACCTGGCAACCTATGAAACCCTGAATATCTCCTGGGCTTCTCAAAGTGGTGAACCACTGGAAACCAAAATTAAAATCATGGAAAGCCATCCCAATGGCACCTTCAGTGCTAAGGGTGTGGCTAGTGTTTGTGTGGAAGACTGGAATAACAGGAAGGAATTTGTGTGTACTGTGACTCACAGGGATCTGCCTTCACCACAGAAGAAATTCATCTCAAAACCCAATGAGGTGCACAAACATCCACCTGCTGTGTACCTGCTGCCACCAGCTCGTGAGCAACTGAACCTGAGGGAGTCAGCCACAGTCACCTGCTTGGTGAAGGGCTTCTCTCCTGCAGACATCAGTGTGCAGTGGCTTCAGAGAGGGCAACTCTTGCCCCAAGAGAAGTATGTGACCAGTGCCCCGATGCCAGAGCCTGGGGCCCCAGGCTTCTACTTTACCCACAGCATCCTGACTGTGACAGAGGAGGAATGGAACTCCGGAGAGACCTATACCTGTGTTGTAGGCCACGAGGCCCTGCCACACCTGGTGACCGAGAGGACCGTGGACAAGTCCACTGGTAAACCCACACTGTACAATGTCTCCCTGATCATGTC TGACACAGGCGGCACCTGCTATTGA<br /> <br /> == Reverse Transcriptase Gene Specific Primers ==<br /> <br /> Cloning from Hybridoma Murine SP2/0-Ag14 (SP2/0)<br /> <br /> Mouse-IgM-RT gacgagggggaagacatttgggaa&amp;nbsp; Published primers ([https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4278175/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4278175/]<br /> &lt;pre&gt;Igκ, AGTGGCCTCACAGGTAT<br /> (ATACCTGTGAGGCCACT, TCEAT)<br /> <br /> IgM, GGTATTCATCTGAACCTTCAAG<br /> (CTTGAAGGTTCAGATGAATACC, LEGSDEY)<br /> <br /> IgG, ACAGTCACTGAGCTGC<br /> (GCAGCTCAGTGACTGT, SSSVTV)<br /> <br /> https://www.sciencedirect.com/science/article/pii/S0161589017304789#tbl0005<br /> <br /> IgG1_2a_2b-CH1-RT, CTGGACAGGGATCCAGAGTTCCA <br /> tggaactctggatccctgtccag<br /> W N S G S L S <br /> <br /> IgG1_Hinge-RT, GGCTTACAACCACAATCCCTG <br /> cagggattgtggttgtaagcc<br /> R D C G C K <br /> <br /> LC-kappa-RT, GGATACAGTTGGTGCAGCATC <br /> gatgctgcaccaactgtatcc<br /> D A A P T V S <br /> <br /> IgG1_2b-CH1-RT GACAGATGGGGGTGTCGTTTTGGC<br /> gccaaaacgacacccccatctgtc<br /> A K T T P P S V <br /> &lt;/pre&gt;<br /> [[Category:Antibodies]]</div> Paul.xu