Skip to main content
This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

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
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
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
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
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
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
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

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