NeuroMab/Trimmer Lab Recombinant mAb and Nanobody 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.
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).
Recombinant Monoclonal Antibody Plasmid Collection
The following plasmids can be used to produce R-mAbs from transfected mammalian cells.
| Addgene ID | Plasmid | Target | Target Species | Host Species | Isotype |
|---|---|---|---|---|---|
| 114473 | Anti-TRIP8b (exon 4) [N212/3R] | TRIP8b (exon 4) | Rat | Mouse | IgG2a |
| 114474 | Anti-Clcn4 [N222/6R] | Clcn4 | Human | Mouse | IgG2a |
| 114475 | Anti-PARIS/ZNF746 [N196/16R] | PARIS/ZNF746 | Human | Mouse | IgG2a |
| 114476 | Anti-Synaptotagmin-3 [N278/19R] | Synaptotagmin-3 | Mouse | Mouse | IgG2a |
| 114477 | Anti-TRPML3/Mucolipin-3 [N268/19R] | TRPML3/Mucolipin-3 | Human | Mouse | IgG2a |
| 114478 | Anti-GABA(A)R, Epsilon [N413/67R] | GABA(A)R, Epsilon | Human | Mouse | IgG2a |
| 114479 | Anti-Ankyrin-R [N388A/10R] | Ankyrin-R | Human | Mouse | IgG2a |
| 114480 | Anti-SUR2B [N323B/20R] | SUR2B | Rat | Mouse | IgG2a |
| 114481 | Anti-QKI-5 [N195A/16R] | QKI-5 | Human | Mouse | IgG2a |
| 114482 | Anti-Synaptotagmin-12 [N277/7R] | Synaptotagmin-12 | Mouse | Mouse | IgG2a |
| 114483 | Anti-GluA1/GluR1 glutamate receptor [N355/1R] | GluA1/GluR1 glutamate receptor | Rat | Mouse | IgG2a |
| 114484 | Anti-Stonin-2 [N346/9R] | Stonin-2 | Human | Mouse | IgG2a |
| 114485 | Anti-NSD3 [N348/82R] | NSD3 | Human | Mouse | IgG2a |
| 114486 | Anti-Flrt3 [N353/13R] | Flrt3 | Mouse | Mouse | IgG2a |
| 114487 | Anti-Botch [N116/14R] | Botch | Rat | Mouse | IgG2a |
| 114488 | Anti-Dopamine D3 receptor [N331/19R] | Dopamine D3 receptor | Human | Mouse | IgG2a |
| 114490 | Anti-Kirrel3, short and long [N321C/49R] | Kirrel3, short and long | Rat | Mouse | IgG2a |
| 114492 | Anti-GFP [N86/38.1R] | GFP | Mouse | IgG2a | |
| 114493 | Anti-NGL-3/LRRC4B [N51/6.2R] | NGL-3/LRRC4B | Rat | Mouse | IgG2a |
| 114494 | Anti-VGlut3 [N34/34.2R] | VGlut3 | Rat | Mouse | IgG2a |
| 114495 | Anti-Foxi3 [N359/28R] | Foxi3 | Mouse | Mouse | IgG2a |
| 114496 | Anti-KCNT2/Slo2.1/Slick K+ channel [N11/33R] | KCNT2/Slo2.1/Slick K+ channel | Mouse | Mouse | IgG2a |
| 114497 | Anti-Cavbeta1 Ca2+ channel [N7/18.1R] | Cavbeta1 Ca2+ channel | Human | Mouse | IgG2a |
| 114499 | Anti-SVOP [N356/23R] | SVOP | Rat | Mouse | IgG2a |
| 114501 | Anti-VGlut2 [N29/29R] | VGlut2 | Rat | Mouse | IgG2a |
| 114502 | Anti-Copper ATPase 1 (Menke's disease protein) [L60/4R] | Copper ATPase 1 (Menke's disease protein) | Human | Mouse | IgG2a |
| 114503 | Anti-LRRK1 [N341/37R] | LRRK1 | Human | Mouse | IgG2a |
| 114505 | Anti-Navbeta4 Na+ channel [N168/6.3R] | Navbeta4 Na+ channel | Rat | Mouse | IgG2a |
| 114506 | Anti-HCN4 [N114/10.1R] | HCN4 | Human | Mouse | IgG2a |
| 114507 | Anti-Neuregulin-HBD (Heparin binding domain, Type I/II) [N120A/9.1R] | Neuregulin-HBD (Heparin binding domain, Type I/II) | Human | Mouse | IgG2a |
| 114508 | Anti-GluN2B/NR2B glutamate receptor [N59/36.1R] | GluN2B/NR2B glutamate receptor | Rat | Mouse | IgG2a |
| 114509 | Anti-Pan-FHF-A [N235/22R] | Pan-FHF-A | Human | Mouse | IgG2a |
| 114510 | Anti-REEP1/2 [N326D/29R] | REEP1/2 | Mouse | Mouse | IgG2a |
| 114511 | Anti-Kv3.1b K+ channel [N16B/8R] | Kv3.1b K+ channel | Rat | Mouse | IgG2a |
| 114512 | Anti-HCN1 [N70/28R] | HCN1 | Rat | Mouse | IgG2a |
| 114513 | Anti-Malin [N85/18.1R] | Malin | Human | Mouse | IgG2a |
| 114514 | Anti-DNMT3L [N117/9.5R] | DNMT3L | Human | Mouse | IgG2a |
| 114515 | Anti-GABA(A)R, Delta [N151/3.3R] | GABA(A)R, Delta | Rat | Mouse | IgG2a |
| 114516 | Anti-Doc2b [N150/21.3R] | Doc2b | Rat | Mouse | IgG2a |
| 114517 | Anti-SNAT1 [N104/32.1R] | SNAT1 | Rat | Mouse | IgG2a |
| 114518 | Anti-Neuroligin-3 [N110/29.4R] | Neuroligin-3 | Rat | Mouse | IgG2a |
| 114519 | Anti-RGS14 [N133/21.2R] | RGS14 | Rat | Mouse | IgG2a |
| 114520 | Anti-Cav3.1 Ca2+ channel [N178A/9.1R] | Cav3.1 Ca2+ channel | Mouse | Mouse | IgG2a |
| 114521 | Anti-Pan-QKI [N147/6.1R] | Pan-QKI | Human | Mouse | IgG2a |
| 114525 | Anti-FGF13/FHF2 [N91/27.1R] | FGF13/FHF2 | Human | Mouse | IgG2a |
| 114527 | Anti-JIP-2/IB-2 [N135/37.2R] | JIP-2/IB-2 | Mouse | Mouse | IgG2a |
| 114529 | Anti-Pan-SAPAP [N127/31.1R] | Pan-SAPAP | Rat | Mouse | IgG2a |
| 114530 | Anti-Neurexin-1-Beta [N170A/1.1R] | Neurexin-1-Beta | Human | Mouse | IgG2a |
| 114533 | Anti-Laforin [N84/37.1R] | Laforin | Human | Mouse | IgG2a |
| 114536 | Anti-GFAP R416WT [N206B/9R] | GFAP R416WT | Human | Mouse | IgG2a |
| 114538 | Anti-FGF12/FHF1 [N94/17R] | FGF12/FHF1 | Human | Mouse | IgG2a |
| 114539 | Anti-MESD [N88/12.1R] | MESD | Mouse | Mouse | IgG2a |
| 114540 | Anti-Olig1 [N149/25.1R] | Olig1 | Mouse | Mouse | IgG2a |
| 114541 | Anti-KCNU1/Slo3 maxi-K+ channel [N2/16R] | KCNU1/Slo3 maxi-K+ channel | Mouse | Mouse | IgG2a |
| 114542 | Anti-LRP4 (cytoplasmic) [N164/6.1R] | LRP4 (cytoplasmic) | Mouse | Mouse | IgG2a |
| 114544 | Anti-GABA(B)R2 [N81/37.1R] | GABA(B)R2 | Human | Mouse | IgG2a |
| 114547 | Anti-Mitofusin-2 [N153/5.1R] | Mitofusin-2 | Mouse | Mouse | IgG2a |
| 114548 | Anti-Mortalin/GRP75 [N52A/42.1R] | Mortalin/GRP75 | Mouse | Mouse | IgG2a |
| 114549 | Anti-GABA(A)R, Beta1 [N96/55.3R] | GABA(A)R, Beta1 | Mouse | Mouse | IgG2a |
| 114551 | Anti-INPP4b [N171/17.4R] | INPP4b | Human | Mouse | IgG2a |
| 114552 | Anti-Pan-Nav1 Na+ channel [N419/40R] | Pan-Nav1 Na+ channel | Human | Mouse | IgG2a |
| 114553 | Anti-Pan-Nav1 Na+ channel [N419/78R] | Pan-Nav1 Na+ channel | Human | Mouse | IgG2a |
| 114554 | Anti-Neuroligin-1 [N97A/31R] | Neuroligin-1 | Rat | Mouse | IgG2a |
| 114555 | Anti-MMACHC [N230/21R] | MMACHC | Human | Mouse | IgG2a |
| 114556 | Anti-SUR1 [N289/16R] | SUR1 | Rat | Mouse | IgG2a |
| 114557 | Anti-TRIP8b (constant) [N212/17R] | TRIP8b (constant) | Rat | Mouse | IgG2a |
| 114558 | Anti-Synaptotagmin-10 [N269/73R] | Synaptotagmin-10 | Mouse | Mouse | IgG2a |
| 114559 | Anti-Ataxin-1, 11NQ [N76/3R] | Ataxin-1, 11NQ | Mouse | Mouse | IgG2a |
| 114560 | Anti-S100A5 [N176A/35.1R] | S100A5 | Rat | Mouse | IgG2a |
| 114561 | Anti-QKI-6 [N182/17.1R] | QKI-6 | Human | Mouse | IgG2a |
| 114562 | Anti-SynCAM4 [N244/5R] | SynCAM4 | Mouse | Mouse | IgG2a |
| 114563 | Anti-TRIP8b (exon 1a/5) [N291C/22R] | TRIP8b (exon 1a/5) | Mouse | Mouse | IgG2a |
| 128612 | Anti-Kv2.1 K+ Channel [D3/71R] | Kv2.1 K+ Channel | Rat | Mouse | IgG2a |
| 128613 | Anti-Kv1.4 K+ channel [K13/31R] | Kv1.4 K+ Channel | Rat | Mouse | IgG2a |
| 128614 | Anti-Kv1.2 K+ Channel [K14/16R] | Kv1.2 K+ Channel | Human | Mouse | IgG2a |
| 128615 | Anti-Kvbeta2/KCNAB2 K+ channel [K17/70R] | Kvbeta2/KCNAB2 K+ Channel | Mouse | Mouse | IgG2a |
| 128616 | Anti-Kv1.1 K+ channel [K20/78R] | Kv1.1 K+ Channel | Rat | Mouse | IgG2a |
| 128617 | Anti-Pan-Kvbeta K+ channel [K25/73R] | Pan-Kvbeta K+ Channel | Mouse | IgG2a | |
| 128618 | Anti-PSD-95 [K28/43R] | PSD-95 | Human | Mouse | IgG2a |
| 128619 | Anti-Kv1.1 K+ channel (External) [K36/15R] | Kv1.1 K+ Channel (External) | Rat | Mouse | IgG2a |
| 128620 | Anti-Kv2.1 K+ channel (External) [K39/25R] | Kv2.1 K+ Channel (External) | Human | Mouse | IgG2a |
| 128621 | K51/1RAnti-SH3GL1/Endophilin A2 [K51/1R] | SH3GL1/Endophilin A2 | Rat | Mouse | IgG2a |
| 128622 | Anti-Kv4.2 K+ Channel (External) [K57/1R] | Kv4.2 K+ Channel (External) | Human | Mouse | IgG2a |
| 128623 | Anti-Pan-Nav channel [K58/35R] | Pan-Nav Channel | Mouse | IgG2a | |
| 128624 | Anti-SAP97 [K64/15R] | SAP97 | Rat | Mouse | IgG2a |
| 128625 | Anti-CASPR/Neurexin IV [K66/38R] | CASPR/Neurexin IV | Rat | Mouse | IgG2a |
| 128626 | Anti-Nav1.1 Na+ channel [K74/71R] | Nav1.1 Na+ channel | Rat | Mouse | IgG2a |
| 128627 | Anti-Kv4.3 K+ channel [K75/41R] | Kv4.3 K+ Channel | Rat | Mouse | IgG2a |
| 128628 | Anti-Kv2.1 K+ Channel [K89/34R] | Kv2.1 K+ Channel | Rat | Mouse | IgG2a |
| 128629 | Anti-WAVE1/SCAR [K91/36R] | WAVE1/SCAR | Human | Mouse | IgG2a |
| 128630 | Anti-PICK1 [L20/8R] | PICK1 | Human | Mouse | IgG2a |
| 128632 | Anti-SynDIG1/Tmem90b [L42/17R] | SynDIG1/Tmem90b | Mouse | Mouse | IgG2a |
| 128633 | Anti-DLX1 [L43/40R] | DLX1 | Human | Mouse | IgG2a |
| 128634 | Anti-Slo1/BKAlpha maxi-K+ channel (Mouse) [L6/23R] | Slo1/BKAlpha maxi-K+ channel | Mouse | Mouse | IgG2a |
| 128635 | Anti-Ankyrin-G (staining) [N106/43R] | Ankyrin-G (staining) | Human | Mouse | IgG2a |
| 128636 | Anti-LRRTM4 [N205B/22R] | LRRTM4 | Human | Mouse | IgG2a |
| 128637 | Anti-Pan-Shank [N23B/49R] | Pan-Shank | Mouse | IgG2a | |
| 128638 | Anti-VGlut1 [N28/9R] | VGlut1 | Rat | Mouse | IgG2a |
| 128639 | Anti-Versican [N351/23R] | Versican | Mouse | Mouse | IgG2a |
| 128640 | Anti-Pan-Ankyrin [N388A/60R] | Pan-Ankyrin | Mouse | IgG2a | |
| 128641 | Anti-Ataxin-1, 11NQ (Mouse) [N76/8R] | Ataxin-1, 11NQ | Mouse | Mouse | IgG2a |
| 140063 | Anti-Kv2.1 K+ channel pS603 [L61/14R] | Kv2.1 K+ channel pS603 | Mouse | Mouse | IgG2a |
| 140064 | Anti-KCNT1/Slo2.2/Slack K+ channel [N3/26R] | KCNT1/Slo2.2/Slack K+ channel | Rt | Mouse | IgG2a |
| 140065 | Anti-SCG10/Stathmin-2 [L5/1R] | SCG10/Stathmin-2 | Rat | Mouse | IgG2a |
| 140066 | Anti-Co-Rest/RCOR1 [K72/8R] | Co-Rest/RCOR1 | Human | Mouse | IgG2a |
| 140068 | Anti-Kv1.6 K+ channel [K19/36R] | Kv1.6 K+ channel | Rat | Mouse | IgG2a |
| 140069 | Anti-Kv1.5 K+ channel [K7/45R] | Kv1.5 K+ channel | Rat | Mouse | IgG2a |
| 140070 | Anti-Kvbeta1.1/KCNAB1 K+ channel [K9/40R] | Kvbeta1.1/KCNAB1 K+ channel | Mouse | Mouse | IgG2a |
| 140071 | Anti-Snapin/SNAPAP [L8/15R] | Snapin/SNAPAP | Human | Mouse | IgG2a |
| 140072 | Anti-Mitofusin-1 [N111/24R] | Mitofusin-1 | Mouse | Mouse | IgG2a |
| 140073 | Anti-Pannexin-2 [N121A/31R] | Pannexin-2 | Rat | Mouse | IgG2a |
| 140074 | Anti-Kir2.2 K+ channel [N124B/38R] | Kir2.2 K+ channel | Rat | Mouse | IgG2a |
| 140075 | Anti-Mad3 [N129A/6R] | Mad3 | Human | Mouse | IgG2a |
| 140076 | Anti-Arl13b [N295B/66R] | Arl13b? | Mouse | Mouse | IgG2a |
| 140077 | Anti-ZNF423 [N328B/37R] | ZNF423 | Human | Mouse | IgG2a |
| 140078 | Anti-Shank3 [N69/46R] | Shank3 | Rat | Mouse | IgG2a |
| 140079 | Anti-GIT2 [N83/48R] | GIT2 | Human | Mouse | IgG2a |
| 140081 | Anti-Neuroligin-4* [N98/7R] | Neuroligin-4* | Mouse | Mouse | IgG2a |
| 149451 | Anti-GluK5/Grik5/KA2 kainate receptor [N279B/27R] | GluK5/Grik5/KA2 kainate receptor | Rat | Mouse | IgG2a |
| 149452 | Anti-Kirrel3, short isoform 3 [N320/48R] | Kirrel3, short isoform 3 | Mouse | Mouse | IgG2a |
| 149453 | Anti-Kv4.2 K+ channel [L28/4R] | Kv4.2 K+ channel | Rat | Mouse | IgG2a |
| 149454 | Anti-BK beta4 K+ channel [L18A/3R] | BK beta4 K+ channel | Mouse | Mouse | IgG2a |
| 149455 | Anti-MMP9 matrix metalloproteinase-9 precursor [L51/82R] | MMP9 matrix metalloproteinase-9 precursor | Rat | Mouse | IgG2a |
| 149456 | Anti-GABA(A)R, Alpha5 [N415/24R] | GABA(A)R, Alpha5 | Human | Mouse | IgG2a |
| 149458 | Anti-Cavbeta2 Ca2+ channel [N8B/1R] | Cavbeta2 Ca2+ channel | Rat | Mouse | IgG2a |
| 164154 | Anti-TRIP8b [N212/7R] | TRIP8b | Rat | Mouse | IgG2a |
| 164155 | Anti-GABA(A)R, alpha6 [N229A/32R] | GABA(A)R, alpha6 | Mouse | Mouse | IgG2a |
| 164156 | Anti-Gamma-protocadherin-C3 [N174B/27R] | Gamma-protocadherin-C3 | Mouse | Mouse | IgG2a |
| 164157 | Anti-Iduna/RNF146 [N201/35R] | Iduna/RNF146 | Mouse | Mouse | IgG2a |
| 164158 | Anti-DNAH7 [N250/21.4R] | DNAH7 | Human | Mouse | IgG2a |
Browse Addgene’s full antibody and nanobody collection here.
Nanobody Plasmid Collection
The following plasmids can be used to produce nanobodies from transfected bacterial cells.
Browse Addgene’s full antibody and nanobody collection here.
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
Trimmer J, et al. 2020. Recombinant Antibodies in Basic Neuroscience Research. Curr Prot Neurosci. 94.1. PMID: 33151027
Dong JX, et al. 2019. A toolbox of nanobodies developed and validated for use as intrabodies and nanoscale immunolabels in mammalian brain neurons. eLife. 8:e48750. PMID: 31566565
Gavira-O'Neill CE, et al. 2020. Development, Screening, and Validation of Camelid-Derived Nanobodies for Neuroscience Research. Curr Prot Neurosci. 94.1. PMID: 33185319