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neuromab icon sm.png NeuroMab/Trimmer Lab Recombinant mAb Plasmid Collection


The UC Davis/NIH NeuroMab Facility together with the laboratory of NeuroMab Director, Dr. James Trimmer, have produced numerous monoclonal antibodies (NeuroMabs) extensively validated for neuroscience research applications. A substantial number of these have been cloned, expressed and validated in recombinant form. The recombinant monoclonal antibodies or R-mAbs have been engineered such that they are a distinct IgG subclass from the conventional mAb to facilitate multiplex labeling with subclass-specific secondary antibodies. The plasmids encoding these R-mAbs have been deposited with Addgene for distribution to the research community. Protocols for the generation R-mAbs plasmids, production of R-mAbs in transfected cells and their validation can be found below.

Schematic of antibody plasmid

With these plasmids researchers will be able to easily and inexpensively generate R-mAbs by transiently transfecting the plasmids into mammalian tissue culture cells (e.g., HEK293, COS, CHO), from which the R-mAbs are secreted into the culture media. For high level expression we recommend using the HEK-2936E cell line engineered by Durocher et al., 2002.

The schematic to the right shows the separate elements of the R-mAb expression plasmid involved in coexpression of light (green) and heavy (blue) chains as driven by two CMV promoters (orange). Hybridoma-derived VL and VH domain PCR products are fused to a joining fragment comprising a κ light chain constant domain (CL) and the κ light chain polyA tail sequences (κ pA), a CMV promoter for heavy chain expression, and an ER signal/leader sequence (L) for translocation of the heavy chain across the ER membrane. PCR-mediated fusion of these three elements is followed by their insertion into the p1316 plasmid that contains an upstream CMV promoter for light chain expression, and an ER signal/leader sequence (L) for translocation of the light chain across the ER membrane. Downstream of the insert is a heavy chain constant domain (CH) that is either γ1 or γ2a depending on the plasmid, followed by the SV40 polyA tail (SV40 pA).


NeuroMab/Trimmer Lab R-mAb Plasmid Collection

The following plasmids can be used to produce R-mAbs from transfected mammalian cells.

Addgene ID Plasmid Target Target Species Antibody Species Isotype
114473 N212/3R TRIP8b (exon 4) Rat Mouse IgG2a
114474 N222/6R Clcn4 Human Mouse IgG2a
114475 N196/16R PARIS/ZNF746 Human Mouse IgG2a
114476 N278/19R Synaptotagmin-3 Mouse Mouse IgG2a
114477 N268/19R TRPML3/Mucolipin-3 Human Mouse IgG2a
114478 N413/67R GABA(A)R, Epsilon Human Mouse IgG2a
114479 N388A/10R Ankyrin-R Human Mouse IgG2a
114480 N323B/20R TRPML3/Mucolipin-3 Human Mouse IgG2a
114481 N195A/16R QKI-5 Human Mouse IgG2a
114482 N277/7R Synaptotagmin-12 Mouse Mouse IgG2a
114483 N355/1R GluA1/GluR1 glutamate receptor Rat Mouse IgG2a
114484 N346/9R Stonin-2 Human Mouse IgG2a
114485 N348/82R NSD3 Human Mouse IgG2a
114486 N353/13R Flrt3 Mouse Mouse IgG2a
114487 N116/14R Botch Rat Mouse IgG2a
114488 N331/19R Dopamine D3 receptor Human Mouse IgG2a
114490 N321C/49R Kirrel3, short and long Rat Mouse IgG2a
114492 N86/38.1R GFP Mouse IgG2a
114493 N51/6.2R NGL-3/LRRC4B Rat Mouse IgG2a
114494 N34/34.2R VGlut3 Rat Mouse IgG2a
114495 N359/28R Foxi3 Mouse Mouse IgG2a
114496 N11/33R KCNT2/Slo2.1/Slick K+ channel Mouse Mouse IgG2a
114497 N7/18.1R Cavbeta1 Ca2+ channel Human Mouse IgG2a
114499 N356/23R SVOP Rat Mouse IgG2a
114501 N29/29R VGlut2 Rat Mouse IgG2a
114502 L60/4R Copper ATPase 1 (Menke's disease protein) Human Mouse IgG2a
114503 N341/37R LRRK1 Human Mouse IgG2a
114505 N168/6.3R Navbeta4 Na+ channel Rat Mouse IgG2a
114506 N114/10.1R HCN4 Human Mouse IgG2a
114507 N120A/9.1R Neuregulin-HBD (Heparin binding domain, Type I/II) Human Mouse IgG2a
114508 N59/36.1R GluN2B/NR2B glutamate receptor Rat Mouse IgG2a
114509 N235/22R Pan-FHF-A Human Mouse IgG2a
114510 N326D/29R REEP1/2 Mouse Mouse IgG2a
114511 N16B/8R Kv3.1b K+ channel Rat Mouse IgG2a
114512 N70/28R HCN1 Rat Mouse IgG2a
114513 N85/18.1R Malin Human Mouse IgG2a
114514 N117/9.5R Kv3.1b K+ channel Rat Mouse IgG2a
114515 N151/3.3R GABA(A)R, Delta Rat Mouse IgG2a
114516 N150/21.3R Doc2b Rat Mouse IgG2a
114517 N104/32.1R SNAT1 Rat Mouse IgG2a
114518 N110/29.4R Neuroligin-3 Rat Mouse IgG2a
114519 N133/21.2R RGS14 Rat Mouse IgG2a
114520 N178A/9.1R Cav3.1 Ca2+ channel Mouse Mouse IgG2a
114521 N147/6.1R Pan-QKI Human Mouse IgG2a
114523 N18/30.1R PSD-93/Chapsyn-110 Rat Mouse IgG2a
114525 N91/27.1R FGF13/FHF2 Human Mouse IgG2a
114527 N135/37.2R JIP-2/IB-2 Mouse Mouse IgG2a
114529 N127/31.1R Pan-SAPAP Rat Mouse IgG2a
114530 N170A/1.1R Neurexin-1-Beta Human Mouse IgG2a
114533 N84/37.1R Laforin Human Mouse IgG2a
114536 N206B/9R GFAP R416WT Human Mouse IgG2a
114538 N94/17R FGF12/FHF1 Human Mouse IgG2a
114539 N88/12.1R MESD Mouse Mouse IgG2a
114540 N149/25.1R Olig1 Mouse Mouse IgG2a
114541 N2/16R KCNU1/Slo3 maxi-K+ channel Mouse Mouse IgG2a
114542 N164/6.1R LRP4 (cytoplasmic) Mouse Mouse IgG2a
114544 N81/37.1R GABA(B)R2 Human Mouse IgG2a
114547 N153/5.1R Mitofusin-2 Mouse Mouse IgG2a
114548 N52A/42.1R Mortalin/GRP75 Mouse Mouse IgG2a
114549 N96/55.3R GABA(A)R, Beta1 Mouse Mouse IgG2a
114550 N93A/49.2R GABA(B)R1 Rat Mouse IgG2a
114551 N171/17.4R INPP4b Human Mouse IgG2a
114552 N419/40R Pan-Nav1 Na+ channel Human Mouse IgG2a
114553 N419/78R Pan-Nav1 Na+ channel Human Mouse IgG2a
114554 N97A/31R Nlgn1 Rat Mouse IgG2a
114555 N230/21R MMACHC Human Mouse IgG2a
114556 N289/16R SUR1 Rat Mouse IgG2a
114557 N212/17R SUR1 Rat Mouse IgG2a
114558 N269/73R Synaptotagmin-10 Mouse Mouse IgG2a
114559 N76/3R Ataxin-1, 11NQ Mouse Mouse IgG2a
114560 N176A/35.1R S100A5 Rat Mouse IgG2a
114561 N182/17.1R QKI-6 Human Mouse IgG2a
114562 N244/5R SynCAM4 Mouse Mouse IgG2a
114563 N291C/22R TRIP8b (exon 1a/5) Mouse Mouse IgG2a
128612 D3/71R Kv2.1 K+ Channel Rat Mouse IgG2a
128613 K13/31R Kv1.4 K+ Channel Rat Mouse IgG2a
128614 K14/16R Kv1.2 K+ Channel Human Mouse IgG2a
128615 K17/70R Kvbeta2/KCNAB2 K+ Channel Mouse Mouse IgG2a
128616 K20/78R Kv1.1 K+ Channel Rat Mouse IgG2a
128617 K25/73R Pan-Kvbeta K+ Channel Mouse IgG2a
128618 K28/43R PSD-95 Human Mouse IgG2a
128619 K36/15R Kv1.1 K+ Channel (External) Rat Mouse IgG2a
128620 K39/25R Kv2.1 K+ Channel (External) Human Mouse IgG2a
128621 K51/1R SH3GL1/Endophilin A2 Rat Mouse IgG2a
128622 K57/1R Kv4.2 K+ Channel (External) Human Mouse IgG2a
128623 K58/35R Pan-Nav Channel Mouse IgG2a
128624 K64/15R SAP97 Rat Mouse IgG2a
128625 K66/38R CASPR/Neurexin IV Rat Mouse IgG2a
128626 K74/71R Nav1.1 Na+ channel Rat Mouse IgG2a
128627 K75/41R Kv4.3 K+ Channel Rat Mouse IgG2a
128628 K89/34R Kv2.1 K+ Channel Rat Mouse IgG2a
128629 K91/36R WAVE1/SCAR Human Mouse IgG2a
128630 L20/8R PICK1 Human Mouse IgG2a
128632 L42/17R SynDIG1/Tmem90b Mouse Mouse IgG2a
128633 L43/40R DLX1 Human Mouse IgG2a
128634 L6/23R Slo1/BKAlpha maxi-K+ channel Mouse Mouse IgG2a
128635 N106/43R Ankyrin-G (staining) Human Mouse IgG2a
128636 N205B/22R LRRTM4 Human Mouse IgG2a
128637 N23B/49R Pan-Shank Mouse IgG2a
128638 N28/9R VGlut1 Rat Mouse IgG2a
128639 N351/23R Versican Mouse Mouse IgG2a
128640 N388A/60R Pan-Ankyrin Mouse IgG2a
128641 N76/8R Ataxin-1, 11NQ Mouse Mouse IgG2a
140063 L61/14R Kv2.1 K+ channel pS603 Rat Mouse IgG2a
140064 N3/26R KCNT1/Slo2.2/Slack K+ channel Rat Mouse IgG2a
140065 L5/1R SCG10/Stathmin-2 Rat Mouse IgG2a
140066 K72/8R Co-Rest/RCOR1 Human Mouse IgG2a
140068 K19/36R Kv1.6 K+ channel Rat Mouse IgG2a
140069 K7/45R Kv1.5 K+ channel Rat Mouse IgG2a
140070 K9/40R Kvbeta1.1/KCNAB1 K+ channel Mouse Mouse IgG2a
140071 L8/15R Snapin/SNAPAP Human Mouse IgG2a
140072 N111/24R Mitofusin-1 Mouse Mouse IgG2a
140073 N121A/31R Pannexin-2 Rat Mouse IgG2a
140074 N124B/38R Kir2.2 K+ channel Rat Mouse IgG2a
140075 N129A/6R Mad3 Human Mouse IgG2a
140076 N295B/66R Arl13b Mouse Mouse IgG2a
140077 N328B/37R ZNF423 Human Mouse IgG2a
140078 N69/46R Shank3 Rat Mouse IgG2a
140079 N83/48R GIT2 Human Mouse IgG2a
140081 N98/7R Neuroligin-4* Mouse Mouse IgG2a
149451 N279B/27R GluK5/Grik5/KA2 kainate receptor Rat Mouse IgG2a
149452 N320/48R Kirrel3, short isoform 3 Mouse Mouse IgG2a
149453 L28/4R Kv4.2 K+ channel Rat Mouse IgG2a
149454 L18A/3R BK beta4 K+ channel Mouse Mouse IgG2a
149455 L51/82R MMP9 matrix metalloproteinase-9 precursor Rat Mouse IgG2a
149456 N415/24R GABA(A)R, Alpha5 Human Mouse IgG2a
149458 N8B/1R Cavbeta2 Ca2+ channel Rat Mouse IgG2a

Protocols

The UC Davis/NIH NeuroMab Facility has several protocols available that may be helpful for working with R-mAbs expressed from the plasmids in this collection.

UC Davis/NIH NeuroMab Protocols

References

Andrews NP, et al. 2019. A toolbox of IgG subclass-switched recombinant monoclonal antibodies for enhanced multiplex immunolabeling of brain. eLife. 8:e43322. PMID: 30667360

Crosnier C, et al. 2010. A rapid and scalable method for selecting recombinant mouse monoclonal antibodies. BMC Biology. 8:76. PMID: 20525357

Durocher Y, et al. 2002. High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res. 30:e9. PMID: 11788735

UC Davis/NIH NeuroMab Publications