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.
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 | IgG1 |
| 114474 | N222/6R | Clcn4 | Human | Mouse | IgG1 |
| 114475 | N196/16R | PARIS/ZNF746 | Human | Mouse | IgG1 |
| 114476 | N278/19R | Synaptotagmin-3 | Mouse | Mouse | IgG1 |
| 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 | IgG2b |
| 114480 | N323B/20R | TRPML3/Mucolipin-3 | Human | Mouse | IgG2a |
| 114481 | N195A/16R | QKI-5 | Human | Mouse | IgG1 |
| 114482 | N277/7R | Synaptotagmin-12 | Mouse | Mouse | IgG1 |
| 114483 | N355/1R | GluA1/GluR1 glutamate receptor | Rat | Mouse | IgG1 |
| 114484 | N346/9R | Stonin-2 | Human | Mouse | IgG2b |
| 114485 | N348/82R | NSD3 | Human | Mouse | IgG1 |
| 114486 | N353/13R | Flrt3 | Mouse | Mouse | IgG1 |
| 114487 | N116/14R | Botch | Rat | Mouse | IgG1 |
| 114488 | N331/19R | Dopamine D3 receptor | Human | Mouse | IgG2a |
| 114490 | N321C/49R | Kirrel3, short and long | Rat | Mouse | IgG2b |
| 114492 | N86/38.1R | GFP | Mouse | IgG2a | |
| 114493 | N51/6.2R | NGL-3/LRRC4B | Rat | Mouse | IgG1 |
| 114494 | N34/34.2R | VGlut3 | Rat | Mouse | IgG1 |
| 114495 | N359/28R | Foxi3 | Mouse | Mouse | IgG1 |
| 114496 | N11/33R | KCNT2/Slo2.1/Slick K+ channel | Mouse | Mouse | IgG1 |
| 114497 | N7/18.1R | Cavbeta1 Ca2+ channel | Human | Mouse | IgG2a |
| 114499 | N356/23R | SVOP | Rat | Mouse | IgG1 |
| 114501 | N29/29R | VGlut2 | Rat | Mouse | IgG1 |
| 114502 | L60/4R | Copper ATPase 1 (Menke's disease protein) | Human | Mouse | IgG2b |
| 114503 | N341/37R | LRRK1 | Human | Mouse | IgG2a |
| 114505 | N168/6.3R | Navbeta4 Na+ channel | Rat | Mouse | IgG1 |
| 114506 | N114/10.1R | HCN4 | Human | Mouse | IgG1 |
| 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 | IgG2b |
| 114509 | N235/22R | Pan-FHF-A | Human | Mouse | IgG2b |
| 114510 | N326D/29R | REEP1/2 | Mouse | Mouse | IgG1 |
| 114511 | N16B/8R | Kv3.1b K+ channel | Rat | Mouse | IgG1 |
| 114512 | N70/28R | HCN1 | Rat | Mouse | IgG1 |
| 114513 | N85/18.1R | Malin | Human | Mouse | IgG1 |
| 114514 | N117/9.5R | Kv3.1b K+ channel | Rat | Mouse | IgG1 |
| 114515 | N151/3.3R | GABA(A)R, Delta | Rat | Mouse | IgG2a |
| 114516 | N150/21.3R | Doc2b | Rat | Mouse | IgG1 |
| 114518 | N110/29.4R | Neuroligin-3 | Rat | Mouse | IgG1 |
| 114519 | N133/21.2R | RGS14 | Rat | Mouse | IgG2a |
| 114520 | N178A/9.1R | Cav3.1 Ca2+ channel | Mouse | Mouse | IgG1 |
| 114521 | N147/6.1R | Pan-QKI | Human | Mouse | IgG2b |
| 114525 | N91/27.1R | FGF13/FHF2 | Human | Mouse | IgG1 |
| 114527 | N135/37.2R | JIP-2/IB-2 | Mouse | Mouse | IgG1 |
| 114529 | N127/31.1R | Pan-SAPAP | Rat | Mouse | IgG2b |
| 114530 | N170A/1.1R | Neurexin-1-Beta | Human | Mouse | IgG1 |
| 114533 | N84/37.1R | Laforin | Human | Mouse | IgG1 |
| 114536 | N206B/9R | GFAP R416WT | Human | Mouse | IgG1 |
| 114538 | N94/17R | FGF12/FHF1 | Human | Mouse | IgG1 |
| 114539 | N88/12.1R | MESD | Mouse | Mouse | IgG2b |
| 114540 | N149/25.1R | Olig1 | Mouse | Mouse | IgG1 |
| 114541 | N2/16R | KCNU1/Slo3 maxi-K+ channel | Mouse | Mouse | IgG1 |
| 114542 | N164/6.1R | LRP4 (cytoplasmic) | Mouse | Mouse | IgG1 |
| 114544 | N81/37.1R | GABA(B)R2 | Human | Mouse | IgG1 |
| 114547 | N153/5.1R | Mitofusin-2 | Mouse | Mouse | IgG2a |
| 114548 | N52A/42.1R | Mortalin/GRP75 | Mouse | Mouse | IgG1 |
| 114549 | N96/55.3R | GABA(A)R, Beta1 | Mouse | Mouse | IgG1 |
| 114550 | N93A/49.2R | GABA(B)R1 | Rat | Mouse | IgG1 |
| 114551 | N171/17.4R | INPP4b | Human | Mouse | IgG1 |
| 114552 | N419/40R | Pan-Nav1 Na+ channel | Human | Mouse | IgG2a |
| 114553 | N419/78R | Pan-Nav1 Na+ channel | Human | Mouse | IgG1 |
| 114554 | N97A/31R | Nlgn1 | Rat | Mouse | IgG1 |
| 114555 | N230/21R | MMACHC | Human | Mouse | IgG1 |
| 114556 | N289/16R | SUR1 | Rat | Mouse | IgG1 |
| 114557 | N212/17R | SUR1 | Rat | Mouse | IgG1 |
| 114558 | N269/73R | Synaptotagmin-10 | Mouse | Mouse | IgG1 |
| 114559 | N76/3R | Ataxin-1, 11NQ | Mouse | Mouse | IgG1 |
| 114560 | N176A/35.1R | S100A5 | Rat | Mouse | IgG1 |
| 114561 | N182/17.1R | QKI-6 | Human | Mouse | IgG1 |
| 114562 | N244/5R | SynCAM4 | Mouse | Mouse | IgG1 |
| 114563 | N291C/22R | TRIP8b (exon 1a/5) | Mouse | Mouse | IgG1 |
| 128612 | D3/71R | Kv2.1 K+ Channel | Rat | Mouse | |
| 128613 | K13/31R | Kv1.4 K+ Channel | Rat | Mouse | IgG1 |
| 128614 | K14/16R | Kv1.2 K+ Channel | Human | Mouse | IgG2b |
| 128615 | K17/70R | Kvbeta2/KCNAB2 K+ Channel | Mouse | Mouse | IgG1 |
| 128616 | K20/78R | Kv1.1 K+ Channel | Rat | Mouse | IgG1 |
| 128617 | K25/73R | Pan-Kvbeta K+ Channel | Mouse | IgG1 | |
| 128618 | K28/43R | PSD-95 | Human | Mouse | IgG2a |
| 128619 | K36/15R | Kv1.1 K+ Channel (External) | Rat | Mouse | IgG2b |
| 128620 | K39/25R | Kv2.1 K+ Channel (External) | Human | Mouse | IgG2a |
| 128621 | K51/1R | SH3GL1/Endophilin A2 | Rat | Mouse | IgG1 |
| 128622 | K57/1R | Kv4.2 K+ Channel (External) | Human | Mouse | IgG1 |
| 128623 | K58/35R | Pan-Nav Channel | Mouse | ||
| 128624 | K64/15R | SAP97 | Rat | Mouse | IgG1 |
| 128625 | K66/38R | CASPR/Neurexin IV | Rat | Mouse | IgG2a |
| 128626 | K74/71R | Nav1.1 Na+ channel | Rat | Mouse | IgG1 |
| 128627 | K75/41R | Kv4.3 K+ Channel | Rat | Mouse | IgG1 |
| 128628 | K89/34R | Kv2.1 K+ Channel | Rat | Mouse | IgG1 |
| 128629 | K91/36R | WAVE1/SCAR | Human | Mouse | IgG1 |
| 128630 | L20/8R | PICK1 | Human | Mouse | IgG2a |
| 128631 | L28/4R | Kv4.2 K+ channel | Rat | Mouse | IgG1 |
| 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 | IgG1 |
| 128635 | N106/43R | Ankyrin-G (staining) | Human | Mouse | IgG1 |
| 128636 | N205B/22R | LRRTM4 | Human | Mouse | IgG1 |
| 128637 | N23B/49R | Pan-Shank | Mouse | IgG1 | |
| 128638 | N28/9R | VGlut1 | Rat | Mouse | IgG1 |
| 128639 | N351/23R | Versican | Mouse | Mouse | IgG1 |
| 128640 | N388A/60R | Pan-Ankyrin | Mouse | IgG1 | |
| 128641 | N76/8R | Ataxin-1, 11NQ | Mouse | Mouse | IgG2b |
| 140076 | N295B/66R | Arl13b | Mouse | Mouse | IgG2a |
| 140079 | N83/48R | GIT2 | Human | Mouse | IgG1 |
| 140081 | N98/7R | Neuroligin-4* | Mouse | Mouse | IgG1 |
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