Key resources table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Purified anti-mouse CD16/32, Clone 93 | Biolegend | CAT# 101330; RRID:AB_2561482 |
| BV711 anti-mouse B220, Clone RA3-6B2 | Biolegend | CAT# 103255; RRID:AB_2563491 |
| AF700 anti-mouse CD19, Clone 6D5 | Biolegend | CAT# 115528; RRID:AB_493735 |
| AF700 anti-mouse CD45.1, Clone A20 | Biolegend | CAT# 110724; RRID:AB_493733 |
| Pacific Blue anti-mouse CD45.1, Clone A20 | Biolegend | CAT# 110722; RRID:AB_492866 |
| BV650 anti-mouse CD45.1, Clone A20 | Biolegend | CAT# 110736; RRID:AB_2562564 |
| AF700 anti-mouse CD45.2, Clone 104 | Biolegend | CAT# 109822; RRID:AB_493731 |
| Pacific Blue anti-mouse CD45.2, Clone 104 | Biolegend | CAT# 109819; RRID:AB_492873 |
| BV650 anti-mouse CD45.2, Clone 104 | Biolegend | CAT# 109836; RRID:AB_2563065 |
| BV510 anti-mouse IgD, Clone 11-26c.2a | Biolegend | CAT# 405723; RRID:AB_2562742 |
| PerCP/Cy5.5 anti-mouse IgD, Clone 11-26c.2a | Biolegend | CAT# 405710; RRID:AB_1575113 |
| Pacific Blue anti-mouse IgD, Clone 11-26c.2a | Biolegend | CAT# 405712; RRID:AB_1937244 |
| Alexa Fluor 647 anti-mouse IgD, Clone 11-26c.2a | Biolegend | CAT# 405708; RRID:AB_893528 |
| BV421 anti-mouse CD138, Clone 281-2 | Biolegend | CAT#142508;RRID:AB_11203544 |
| AF647 anti-mouse GL7, Clone GL7 | Biolegend | CAT# 144606; RRID:AB_2562185 |
| PerCP/Cy5.5 anti-mouse GL7, Clone GL7 | Biolegend | CAT# 144610; RRID:AB_2562979 |
| PE/Cy7anti-mouse CD38, Clone 90 | Biolegend | CAT# 102718; RRID:AB_2275531 |
| PerCP/Cy5.5 anti-mouse CD73, Clone TY/11.8 | Biolegend | CAT#127214;RRID:AB_11219403 |
| BV605 anti-mouse CD80, Clone 16-10A1 | Biolegend | CAT#104729;RRID:AB_11126141 |
| BV605 anti-mouse CD86, Clone GL-1 | Biolegend | CAT#105037;RRID:AB_11204429 |
| FITC anti-mouse CD86, Clone GL-1 | Biolegend | CAT# 105005; RRID:AB_313148 |
| Biotin anti-mouse CXCR4, Clone 2B11/CXCR4 | BD Biosciences | CAT# 551968; RRID:AB_394307 |
| PE anti-mouse CXCR4, Clone 2B11/CXCR4 | BD Biosciences | CAT# 551966; RRID:AB_394305 |
| PE anti-mouse CCR9, Clone 9B1 | Biolegend | CAT# 129708; RRID:AB_2073249 |
| Biotin anti-mouse LPAM-1, Clone DATK32 | Biolegend | CAT#120612;RRID:AB_11203892 |
| APC anti-mouse IgM, Clone II/41 | BD Biosciences | CAT# 550676; RRID:AB_398464 |
| FITC anti-mouse IgM, Clone II/41 | BD Biosciences | CAT# 553437; RRID:AB_394857 |
| Biotin anti-mouse IgM, Clone II/41 | BD Biosciences | CAT# 553436;RRID:AB_394856 |
| FITC anti-mouse IgMa, Clone MA-69 | Biolegend | CAT# 408606; RRID:AB_940541 |
| PE anti-mouse IgMa, Clone MA-69 | Biolegend | CAT# 408608; RRID: AB_940545 |
| FITC anti-mouse IgMb, Clone AF6-78 | Biolegend | CAT# 406206; RRID: AB_315039 |
| PE anti-mouse IgMb, Clone AF6-78 | Biolegend | CAT# 406208; RRID: AB_315041 |
| FITC anti-mouse IgG1, Clone RMG1-1 | Biolegend | CAT# 406605; RRID: AB_493292 |
| PE ant-mouse IgG1, Clone RMG1-1 | Biolegend | CAT# 406607; RRID:AB_10551439 |
| FITC anti-mouse IgG2b, Clone RMG2b-1 | Biolegend | CAT# 406705; RRID: AB_493296 |
| PE anti-mouse IgG2b, Clone RMG2b-1 | Biolegend | CAT# 406708; RRID:AB_2563381 |
| Biotin anti-mouse IgG3, Clone RMG3-1 | Biolegend | CAT# 406803; RRID: AB_315070 |
| FITC anti-mouse IgA | Southern Biotech | CAT# 1040-02; RRID: AB_2794370 |
| PE anti-mouse IgA | Southern Biotech | CAT# 1040-09; RRID: AB_2794375 |
| Biotin anti-mouse IgA | Southern Biotech | CAT# 1040-08; RRID: AB_2794374 |
| Biotin anti-active Caspase 3 | BD Biosciences | CAT# 550557; RRID: AB_393750 |
| AF700 anti-mouse Kappa | Biolegend | CAT# 409508; RRID:AB_2563583 |
| APC anti-mouse CD22, Clone OX-97 | Biolegend | CAT# 126110; RRID:AB_2561630 |
| PerCP/Cy5.5 anti-mouse I-A/I-E, Clone M5/114.15.2 | Biolegend | CAT# 107626; RRID:AB_2191071 |
| PerCP/CY5.5 anti-mouse CD79b, Clone HM79-12 | Biolegend | CAT# 132810; RRID:AB_2632918 |
| PE/Cy7 anti-mouse CD69, Clone H1.2F3 | Biolegend | CAT# 104512; RRID: AB_493564 |
| AF700 anti-mouse CD4, Clone RM4-4 | Biolegend | CAT# 116022; RRID:AB_2715958 |
| PE anti-mouse Valpha2 TCR, Clone B20.1 | BD Biosciences | CAT# 553289; RRID: AB_394760 |
| Pacific Blue anti-mouse TCRbeta, Clone H57-597 | Biolegend | CAT# 109226; RRID:AB_1027649 |
| PE/Cy7anti-mouse Fas, Clone Jo2 | BD Biosciences | CAT# 557653; RRID: AB_396768 |
| FITC Annexin V | BD Biosciences | CAT# 556419; RRID: AB_2665412 |
| PE anti-BTK (pY223), Clone N35-86 | BD Biosciences | CAT# 562753; RRID:AB_2737769 |
| AF647 anti-Syk (pY352), Clone 17A/P-ZAP70 | BD Biosciences | CAT# 557817; RRID: AB_396884 |
| PE anti-phosphotyrosine, Clone PY20 | Biolegend | CAT# 309310; RRID:AB_2572196 |
| AF647 anti-mouse CD98, Clone RL388 | Biolegend | CAT#128210; RRID:AB_2254922 |
| Biotin anti-mouse IgD , Clone 11-26c.2a | Biolegend | CAT# 405734; RRID: AB_2563344 |
| Biotin anti-mouse CD138, Clone 281-2 | Biolegend | CAT# 142512; RRID: AB_2561981 |
| Biotin anti-mouse IgG2b, Clone RMG2b-1 | Biolegend | CAT# 406704; RRID: AB_315067 |
| Biotin anti-mouse CD8a, Clone 53-6.7 | Biolegend | CAT# 100704; RRID: AB_312743 |
| Biotin anti-mouse CD4, Clone GK1.5 | Biolegend | CAT# 100404; RRID: AB_312689 |
| Biotin anti-mouse IgG1, Clone RMG1-1 | Biolegend | CAT# 406604; RRID: AB_315063 |
| Biotin anti-mouse IgMa, Clone MA-69 | Biolegend | CAT# 408603; RRID: AB_940535 |
| Purified anti-mouse IgMa, Clone MA-69 | Biolegend | CAT# 408602; RRID: AB_940549 |
| Biotin anti-mouse IgMb, Clone AF6-78 | Biolegend | CAT# 406204; RRID: AB_315037 |
| Purified anti-mouse IgMb, Clone AF6-78 | Biolegend | CAT# 406202; RRID: AB_315035 |
| Unlabeled Goat anti-mouse IgA | Southern Biotech | CAT# 1040-01; RRID: AB_2314669 |
| Unlabeled Goat anti-mouse IgG2b | Southern Biotech | CAT# 1090-01; RRID: AB_2794517 |
| Unlabeled Goat anti-mouse IgM | Southern Biotech | CAT# 1020-01; RRID: AB_2794197 |
| Biotin anti-mouse GL7, Clone GL7 | Biolegend | CAT# 144616; RRID: AB_2721505 |
| Rabbit Anti-RFP | Rockland | CAT# 600401379; RRID: AB_11182807 |
| AF647 anti-mouse CD21/35, Clone 7E9 | Biolegend | CAT#123424; RRID: AB_2629577 |
| Mouse Anti-Phosphotyrosine, Clone 4g10 | Millipore Sigma | CAT# 05-321; RRID: AB_309678 |
| Mouse Anti beta-Actin, Clone AC-74 | Millipore Sigma | CAT# A5316; RRID: AB_476743 |
| HRP linked anti-mouse IgG | Cell Signaling | CAT# 7076; RRID: AB_330924 |
| HRP conjugated anti-mouse Phospho-Tyrosine (p-Tyr-100) | Cell Signaling | CAT# 5465; RRID: AB_10694719 |
| Purified mouse anti-actin, Clone C4/actin | BD Biosciences | CAT# 612657; RRID: AB_399901 |
| Goat polyclonal anti-mouse IgD | Nordic Immunology | CAT# GAM/IgD(Fc)/7S |
| Rabbit polyclonal anti-RFP | Rockland | CAT# 601-401-379 |
| Donkey anti-rabbit Cy3 | Jackson Immunoresearch | CAT# 711-165-152; RRID: AB 2307443 |
| Donkey anti-goat AF488 | Jackson Immunoresearch | CAT# 705-545-003; RRID: AB_2340428 |
| Human TruStain FcX | Biolegend | CAT# 422302; RRID:AB_2818986 |
| PE/Cy7 anti-human CD19, Clone HIB19 | Biolegend | CAT# 560728; RRID:AB_1727438 |
| BV605 anti-human CD27, Clone O323 | Biolegend | CAT# 302830; RRID:AB_2561450 |
| APC anti-human CD10, Clone HI10a | Biolegend | CAT# 312210; RRID:AB_314921 |
| APC/Cy7 anti-human CD10, Clone HI10a | Biolegend | CAT# 312212; RRID:AB_2146550 |
| PerCP/Cy5.5 anti-human CD138, Clone MI15 | Biolegend | CAT#356509; RRID:AB_2561898 |
| PE Goat F(ab’)2 Anti-human IgM | Southern Biotech | CAT# 2022-09; RRID: AB_2795614 |
| PE Goat F(ab’)2 Anti-human IgA | Southern Biotech | CAT# 2052-09; RRID: AB_2687523 |
| Unlabeled Goat F(ab’)2 Anti-human IgM | Southern Biotech | CAT# 2022-01; RRID: AB_2795610 |
| Unlabeled Goat F(ab’)2 Anti-human IgA | Southern Biotech | CAT# 2052-01; RRID: AB_2795709 |
| FITC Goat F(ab’)2 Anti-human IgG | Thermo Fisher | CAT#AHI1308;RRID:AB_1500723 |
| Bacterial strains | ||
| S. enterica strain SL1344 ΔaroA OVA | Gift from M. Bogunovic | N/A |
| Biological samples | ||
| Frozen human tonsils | Gift from E. Clark | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| AF488 Streptavidin | Biolegend | CAT# 405235 |
| PerCP/Cy5.5 Streptavidin | Biolegend | CAT# 405214 |
| BV605 Streptavidin | Biolegend | CAT# 405229 |
| BV650 Streptavidin | Biolegend | CAT# 405231 |
| PE/Cy7Streptavidin | eBioscience | CAT# 25-4317-82 |
| eFluor780 Live/Dead | BD Biosciences | CAT# 4302687 |
| EF450 Live/Dead | eBioscience | CAT# 65-0863-18 |
| DAPI | Thermo Fisher | CAT# 1306 |
| Cholera Toxin | Sigma | CAT# C8052-2MG |
| 16% PFA | Electrom microscopy sciences | CAT# 15710 |
| DNase I | Sigma | CAT# DN25 |
| Collagenase IV | Worthington | CAT# LS004189 |
| Percoll | GE Healthcare | CAT# 17089101 |
| FTY-720 | Selleck Chemicals | CAT# S5002 |
| Isotonic saline | Ricca Chemical | CAT# 7210-16 |
| Ibrutinib | Selleckchem | CAT# S2680 |
| R406 | Invivogen | CAT# inhr406 |
| Fluo 3 AM | Invitrogen | CAT# F23915 |
| Fura Red | Invitrogen | CAT# F3021 |
| 5-Fluorouracil | Sigma | CAT# F6627 |
| Recombinant Murine IL-3 | Peprotech | CAT# 213-13 |
| Recombinant Murine IL-6 | Peprotech | CAT# 216-16 |
| Recombinant Murine stem cell factor | Peprotech | CAT# 250-03 |
| Lipofectamine 2000 | Invitrogen | CAT# 11668-019 |
| RIPA buffer | Thermo Fisher | CAT# 89900 |
| 4x Laemmli sample buffer | Bio-Rad | CAT# 1610747 |
| Phosphatase inhibitor | Thermo Fisher | CAT# 1861281 |
| BSA | Sigma | CAT# A2153 |
| 10% Mini-PROTEAN® TGX™ Precast Protein Gels | Biorad | CAT# 4561033 |
| Protein Standard | BioRad | CAT# 161-0374 |
| Trans-Blot Turbo Mini 0.2 μm PVDF Transfer Packs | BioRad | CAT# 1704156 |
| ECL prime western blotting detection | GE Healthcare | CAT# RPN2232 |
| SuperSignal West Pico Plus substrate | Thermo Fisher | CAT# 34577 |
| 4 to 12% NuPAGE Bis-Tris gels | Invitrogen | CAT# NP0322BOX |
| Restore PLUS Western blot stripping buffer | Thermo Fisher | CAT# 46430 |
| Napthol AS-MX Phosphate | Sigma | CAT# N4875 |
| N-N-Dimethylformamide | Fisher | CAT# D119-500 |
| Levamisole | Sigma | CAT# L9756 |
| Fast Blue B salt | Sigma | CAT# D9805 |
| SIGMAFAST 3,3’-Diaminobenzidine tablets | Sigma | CAT# D4293 |
| Critical commercial assays | ||
| BRDU FITC kit | BD Biosciences | CAT# 559619 |
| BD Cytofix/Cytoperm | BD Biosciences | CAT# 554714 |
| EasySep Streptavidin RapidSpheres Kit | Stemcell Technologies | CAT# 19860A |
| SYBR green master mix | Biorad | CAT# 1725270 |
| Fluoromount-G | Southern Biotech | CAT# 0100-01 |
| Elisa/Elispot diluent solution | Invitrogen | CAT# 004202 |
| Stop Solution for TMB Substrate | Biolegend | CAT# 423001 |
| Deposited data | ||
| N/A | ||
| Experimental models: Cell lines | ||
| NB-21 | Gift from G. Kelsoe | Kuraoka et al., 2016 |
| I29μ | Gift from C. Schrader | N/A |
| CH12 | Gift from J. Muppidi | N/A |
| Platinum-E (Plat-E) retroviral packaging cell line | Gift from S. Schwab | N/A |
| Experimental models: Organisms/strains | ||
| C57BL/6J (CD45.2) | The Jackson Laboratory | Strain# 000664 ; RRID: IMSR_JAX:000664 |
| Ly5.2 (CD45.1) congenic B6.SJL-Ptprca Pepcb/BoyJ | The Jackson Laboratory | Strain# 002014; RRID: IMSR_JAX:002014 |
| Rosa26-flox-stop-flox-tdTomato | The Jackson Laboratory | Strain# 007914; RRID: IMSR_JAX:007914 |
| Aicda-cre | The Jackson Laboratory | Strain# 007770; RRID: IMSR_JAX:007770 |
| B6.MRL-Faslpr | The Jackson Laboratory | Strain# 000482; RRID: IMSR_JAX:000482 |
| B6J.Cg-Gt(ROSA)26Sortm95.1(CAG-GCaMPgf)Hze/MwarJ (gCAMP) | The Jackson Laboratory | Strain# 028865; RRID: IMSR_JAX:028865 |
| B6.Cg-Tg(TcraTcrb)425Cbn/J (OTII) | The Jackson Laboratory | Strain# 004194; RRID: IMSR_JAX:004194 |
| B6.Cg-Rag2tm1.1Cgn/J | The Jackson Laboratory | Strain# 008449; RRID: IMSR_JAX:008449 |
| B6.129SC-Ighmtm1Cgn/J | The Jackson Laboratory | Strain# 002288; RRID: IMSR_JAX:002288 |
| Iga −/− | Barton Lab | N/A |
| IgMa | Reboldi Lab | N/A |
| Rosa26-INDIA | Mayer Lab | N/A |
| cMYC-GFP | Bannard Lab | N/A |
| Oligonucleotides | ||
| N/A | ||
| Recombinant DNA | ||
| MSCV-IRES-blc2 | Gift from J. Muppidi | |
| Software and algorithms | ||
| FlowJo | Tree Star | https://www.flowjo.com |
| FACSDiva | BD | https://www.bdbiosciences.com/en-us/products/software/instrument-software/bd-facsdiva-software |
| Prism | GraphPad Software | https://www.graphpad.com |
| ZEN 3.1 | Carl Zeiss Microscopy | https://www.zeiss.com/microscopy/en/products/software/zeiss-zen.html |
| ImageJ | NIH | https://imagej.net/ij/index.html |
| Adobe Illustrator | Adobe Systems | https://www.adobe.com |
| Biorender | Biorender | https://www.biorender.com |
| Other | ||