Enzyme-linked Immunosorbent Assay:Article Title: Regulation of Foxo1 expression is critical for central B cell tolerance and allelic exclusion.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-B220 BioLegend Clone# RA3-6B2; RRID:AB_11203907 Anti-IgD BD Biosciences Clone# 11-26c-2a; RRID:AB_2738723 Anti-IgM eBioscience Clone# eB121-15F9; RRID:AB_10671539 Anti-IgM, polyclonal (for ELISA) SouthernBiotech Cat# 1021-01; RRID:AB_2687524 Anti-IgG1 (for ELISA) BioLegend or BD Biosciences Clone# A85-3; RRID:AB_394863 Anti-IgG2a (for ELISA) BioLegend or BD Biosciences Clone# RMG2a-62; RRID:AB_2563481 Anti-IgG2b (for ELISA) BioLegend or BD Biosciences Clone# RMG2b-1; RRID:AB_315065 Anti-IgG3 (for ELISA) BioLegend or BD Biosciences Clone# R2-38; RRID:AB_394841 Anti-CD43 BD Biosciences Clone# 1G10; RRID:AB_394748 Anti-CD21 BioLegend Clone# 7E9; RRID:AB_1953277 Anti-CD23 BD Biosciences Clone# B3B4; RRID:AB_2738526 Anti-CD24 BioLegend Clone# M1/69; RRID:AB_1595596 Anti-CD19 BD Biosciences Clone# 1D3; RRID:AB_2737915 Anti-Iglambda BioLegend Clone# RML-42; RRID:AB_961363 Anti-Iglambda SouthernBiotech Cat# 1060-02; RRID:AB_2794390 Anti-Igkappa BD Biosciences Clone# 187.1; RRID:AB_1727535 Anti-IL-7R BioLegend Clone# A7R34; RRID:AB_1937252 Anti-BAFFR R&D Systems Clone# BAF1357; RRID:AB_2271820 Anti-CXCR4 BioLegend Clone# L276F12; RRID:AB_2562785 Anti-CD93 eBioscience Clone# AA4.1; RID:AB_469466 Anti-CD1d BioLegend Clone# 1B1; RRID:AB_10683167 Anti-pH2AX BioLegend Clone# 2F3; RRID:AB_528919 Anti-pAKT (T308) Cell Signaling Technology Clone# C31E5E; RRID:AB_2255933 Anti-Foxo1 Cell Signaling Technology Clone# C29H4; RRID:AB_2106495 Anti-tri-methyl-histone H3 Cell Signaling Technology Clone# C36B11; RRID:AB_2799182 Anti-rabbit IgG-FITC SouthernBiotech Cat# 4050-02; RRID:AB_2795952 Anti-rabbit IgG-DyLight 649 BioLegend Cat# 406406; RRID:AB_1575135 Anti-3-83Ig(H + L) Produced in-house, University of Colorado Anschutz Medical Campus Clone# 54.1 Anti-B220 Microbeads Miltenyi Biotec Cat# 130-049-501 Anti-FITC Microbeads Miltenyi Biotec Cat# 130-058-701 Chemicals, peptides, and recombinant proteins Phosphate substrate Millipore Sigma #P5744 Streptavidin-AP SouthernBiotech #7105-05 Streptavidin-APC-Cy7 BD Biosciences #554063 BSA Fisher Scientific #BP1600-100 Critical commercial assays Zombie UV Fixable Viability Kit BioLegend #423108 eBioscienceTM Foxp3/Transcription Factor Staining Buffer Set eBioscience #00-5523-00 BioLegend Fixation Buffer BioLegend #420801 APC BrdU Flow Kit BD Pharmingen #552598 RNeasy PlusMini Kit Qiagen #74134 SuperScript IV VILO Master Mix Thermo Fisher Scientific #11756050 (Continued on next page) 16 Cell Reports 43, 114283, June 25, 2024 .. REAGENT or RESOURCE SOURCE IDENTIFIER Taqman Gene Expression Assay, Rag1, Mm01270936_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Rag2, Mm00501300_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Foxo1, Mm00490671_m1 Thermo Fisher Scientific #4331182 Taqman Gene Expression Assay, Cxcr4, Mm01996749_s1 Thermo Fisher Scientific #4453320 Taqman Gene Expression Assay, Cd79b, Mm00434143_m1 Thermo Fisher Scientific #4331182 Taqman Universal PCR Master Mix Applied Biosystems #4364338 Experimental models: Organisms/strains 3-83Igi,H-2d mice Braun et al74 Jackson Laboratories #011074 3-83Igi,H-2b mice Halverson et al26 N/A 3-83Igi,Rosa26-P110*,mb1-Cre mice Greaves et al31 N/A Rosa26-P110* mice Srinivasan et al55 Jackson Laboratories #012343 Mb1-Cre mice Hobeika et al75 Jackson Laboratories #029412 3-83Igi,Foxo1fl/fl,mb1-Cre mice This paper N/A Foxo1fl/fl mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-Foxo1-AAA,mb1-Cre mice This paper N/A Foxo1-AAA mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-P110*/Foxo1-AAA,mb1-Cre mice This paper N/A Software and algorithms Flowjo v10.9 BD https://www.flowjo.com/ GraphPad Prism 10 GraphPad https://www.graphpad.com/ QuantStudio 7 Flex Real-Time PCR System software Thermo Fisher Scientific https://www.thermofisher.com/ Other Fetal Bovine Serum Gemini #100106, Lot#050A07H00K
Produced:Article Title: Regulation of Foxo1 expression is critical for central B cell tolerance and allelic exclusion.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-B220 BioLegend Clone# RA3-6B2; RRID:AB_11203907 Anti-IgD BD Biosciences Clone# 11-26c-2a; RRID:AB_2738723 Anti-IgM eBioscience Clone# eB121-15F9; RRID:AB_10671539 Anti-IgM, polyclonal (for ELISA) SouthernBiotech Cat# 1021-01; RRID:AB_2687524 Anti-IgG1 (for ELISA) BioLegend or BD Biosciences Clone# A85-3; RRID:AB_394863 Anti-IgG2a (for ELISA) BioLegend or BD Biosciences Clone# RMG2a-62; RRID:AB_2563481 Anti-IgG2b (for ELISA) BioLegend or BD Biosciences Clone# RMG2b-1; RRID:AB_315065 Anti-IgG3 (for ELISA) BioLegend or BD Biosciences Clone# R2-38; RRID:AB_394841 Anti-CD43 BD Biosciences Clone# 1G10; RRID:AB_394748 Anti-CD21 BioLegend Clone# 7E9; RRID:AB_1953277 Anti-CD23 BD Biosciences Clone# B3B4; RRID:AB_2738526 Anti-CD24 BioLegend Clone# M1/69; RRID:AB_1595596 Anti-CD19 BD Biosciences Clone# 1D3; RRID:AB_2737915 Anti-Iglambda BioLegend Clone# RML-42; RRID:AB_961363 Anti-Iglambda SouthernBiotech Cat# 1060-02; RRID:AB_2794390 Anti-Igkappa BD Biosciences Clone# 187.1; RRID:AB_1727535 Anti-IL-7R BioLegend Clone# A7R34; RRID:AB_1937252 Anti-BAFFR R&D Systems Clone# BAF1357; RRID:AB_2271820 Anti-CXCR4 BioLegend Clone# L276F12; RRID:AB_2562785 Anti-CD93 eBioscience Clone# AA4.1; RID:AB_469466 Anti-CD1d BioLegend Clone# 1B1; RRID:AB_10683167 Anti-pH2AX BioLegend Clone# 2F3; RRID:AB_528919 Anti-pAKT (T308) Cell Signaling Technology Clone# C31E5E; RRID:AB_2255933 Anti-Foxo1 Cell Signaling Technology Clone# C29H4; RRID:AB_2106495 Anti-tri-methyl-histone H3 Cell Signaling Technology Clone# C36B11; RRID:AB_2799182 Anti-rabbit IgG-FITC SouthernBiotech Cat# 4050-02; RRID:AB_2795952 Anti-rabbit IgG-DyLight 649 BioLegend Cat# 406406; RRID:AB_1575135 Anti-3-83Ig(H + L) Produced in-house, University of Colorado Anschutz Medical Campus Clone# 54.1 Anti-B220 Microbeads Miltenyi Biotec Cat# 130-049-501 Anti-FITC Microbeads Miltenyi Biotec Cat# 130-058-701 Chemicals, peptides, and recombinant proteins Phosphate substrate Millipore Sigma #P5744 Streptavidin-AP SouthernBiotech #7105-05 Streptavidin-APC-Cy7 BD Biosciences #554063 BSA Fisher Scientific #BP1600-100 Critical commercial assays Zombie UV Fixable Viability Kit BioLegend #423108 eBioscienceTM Foxp3/Transcription Factor Staining Buffer Set eBioscience #00-5523-00 BioLegend Fixation Buffer BioLegend #420801 APC BrdU Flow Kit BD Pharmingen #552598 RNeasy PlusMini Kit Qiagen #74134 SuperScript IV VILO Master Mix Thermo Fisher Scientific #11756050 (Continued on next page) 16 Cell Reports 43, 114283, June 25, 2024 .. REAGENT or RESOURCE SOURCE IDENTIFIER Taqman Gene Expression Assay, Rag1, Mm01270936_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Rag2, Mm00501300_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Foxo1, Mm00490671_m1 Thermo Fisher Scientific #4331182 Taqman Gene Expression Assay, Cxcr4, Mm01996749_s1 Thermo Fisher Scientific #4453320 Taqman Gene Expression Assay, Cd79b, Mm00434143_m1 Thermo Fisher Scientific #4331182 Taqman Universal PCR Master Mix Applied Biosystems #4364338 Experimental models: Organisms/strains 3-83Igi,H-2d mice Braun et al74 Jackson Laboratories #011074 3-83Igi,H-2b mice Halverson et al26 N/A 3-83Igi,Rosa26-P110*,mb1-Cre mice Greaves et al31 N/A Rosa26-P110* mice Srinivasan et al55 Jackson Laboratories #012343 Mb1-Cre mice Hobeika et al75 Jackson Laboratories #029412 3-83Igi,Foxo1fl/fl,mb1-Cre mice This paper N/A Foxo1fl/fl mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-Foxo1-AAA,mb1-Cre mice This paper N/A Foxo1-AAA mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-P110*/Foxo1-AAA,mb1-Cre mice This paper N/A Software and algorithms Flowjo v10.9 BD https://www.flowjo.com/ GraphPad Prism 10 GraphPad https://www.graphpad.com/ QuantStudio 7 Flex Real-Time PCR System software Thermo Fisher Scientific https://www.thermofisher.com/ Other Fetal Bovine Serum Gemini #100106, Lot#050A07H00K
Recombinant:Article Title: Regulation of Foxo1 expression is critical for central B cell tolerance and allelic exclusion.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-B220 BioLegend Clone# RA3-6B2; RRID:AB_11203907 Anti-IgD BD Biosciences Clone# 11-26c-2a; RRID:AB_2738723 Anti-IgM eBioscience Clone# eB121-15F9; RRID:AB_10671539 Anti-IgM, polyclonal (for ELISA) SouthernBiotech Cat# 1021-01; RRID:AB_2687524 Anti-IgG1 (for ELISA) BioLegend or BD Biosciences Clone# A85-3; RRID:AB_394863 Anti-IgG2a (for ELISA) BioLegend or BD Biosciences Clone# RMG2a-62; RRID:AB_2563481 Anti-IgG2b (for ELISA) BioLegend or BD Biosciences Clone# RMG2b-1; RRID:AB_315065 Anti-IgG3 (for ELISA) BioLegend or BD Biosciences Clone# R2-38; RRID:AB_394841 Anti-CD43 BD Biosciences Clone# 1G10; RRID:AB_394748 Anti-CD21 BioLegend Clone# 7E9; RRID:AB_1953277 Anti-CD23 BD Biosciences Clone# B3B4; RRID:AB_2738526 Anti-CD24 BioLegend Clone# M1/69; RRID:AB_1595596 Anti-CD19 BD Biosciences Clone# 1D3; RRID:AB_2737915 Anti-Iglambda BioLegend Clone# RML-42; RRID:AB_961363 Anti-Iglambda SouthernBiotech Cat# 1060-02; RRID:AB_2794390 Anti-Igkappa BD Biosciences Clone# 187.1; RRID:AB_1727535 Anti-IL-7R BioLegend Clone# A7R34; RRID:AB_1937252 Anti-BAFFR R&D Systems Clone# BAF1357; RRID:AB_2271820 Anti-CXCR4 BioLegend Clone# L276F12; RRID:AB_2562785 Anti-CD93 eBioscience Clone# AA4.1; RID:AB_469466 Anti-CD1d BioLegend Clone# 1B1; RRID:AB_10683167 Anti-pH2AX BioLegend Clone# 2F3; RRID:AB_528919 Anti-pAKT (T308) Cell Signaling Technology Clone# C31E5E; RRID:AB_2255933 Anti-Foxo1 Cell Signaling Technology Clone# C29H4; RRID:AB_2106495 Anti-tri-methyl-histone H3 Cell Signaling Technology Clone# C36B11; RRID:AB_2799182 Anti-rabbit IgG-FITC SouthernBiotech Cat# 4050-02; RRID:AB_2795952 Anti-rabbit IgG-DyLight 649 BioLegend Cat# 406406; RRID:AB_1575135 Anti-3-83Ig(H + L) Produced in-house, University of Colorado Anschutz Medical Campus Clone# 54.1 Anti-B220 Microbeads Miltenyi Biotec Cat# 130-049-501 Anti-FITC Microbeads Miltenyi Biotec Cat# 130-058-701 Chemicals, peptides, and recombinant proteins Phosphate substrate Millipore Sigma #P5744 Streptavidin-AP SouthernBiotech #7105-05 Streptavidin-APC-Cy7 BD Biosciences #554063 BSA Fisher Scientific #BP1600-100 Critical commercial assays Zombie UV Fixable Viability Kit BioLegend #423108 eBioscienceTM Foxp3/Transcription Factor Staining Buffer Set eBioscience #00-5523-00 BioLegend Fixation Buffer BioLegend #420801 APC BrdU Flow Kit BD Pharmingen #552598 RNeasy PlusMini Kit Qiagen #74134 SuperScript IV VILO Master Mix Thermo Fisher Scientific #11756050 (Continued on next page) 16 Cell Reports 43, 114283, June 25, 2024 .. REAGENT or RESOURCE SOURCE IDENTIFIER Taqman Gene Expression Assay, Rag1, Mm01270936_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Rag2, Mm00501300_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Foxo1, Mm00490671_m1 Thermo Fisher Scientific #4331182 Taqman Gene Expression Assay, Cxcr4, Mm01996749_s1 Thermo Fisher Scientific #4453320 Taqman Gene Expression Assay, Cd79b, Mm00434143_m1 Thermo Fisher Scientific #4331182 Taqman Universal PCR Master Mix Applied Biosystems #4364338 Experimental models: Organisms/strains 3-83Igi,H-2d mice Braun et al74 Jackson Laboratories #011074 3-83Igi,H-2b mice Halverson et al26 N/A 3-83Igi,Rosa26-P110*,mb1-Cre mice Greaves et al31 N/A Rosa26-P110* mice Srinivasan et al55 Jackson Laboratories #012343 Mb1-Cre mice Hobeika et al75 Jackson Laboratories #029412 3-83Igi,Foxo1fl/fl,mb1-Cre mice This paper N/A Foxo1fl/fl mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-Foxo1-AAA,mb1-Cre mice This paper N/A Foxo1-AAA mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-P110*/Foxo1-AAA,mb1-Cre mice This paper N/A Software and algorithms Flowjo v10.9 BD https://www.flowjo.com/ GraphPad Prism 10 GraphPad https://www.graphpad.com/ QuantStudio 7 Flex Real-Time PCR System software Thermo Fisher Scientific https://www.thermofisher.com/ Other Fetal Bovine Serum Gemini #100106, Lot#050A07H00K
Staining:Article Title: Regulation of Foxo1 expression is critical for central B cell tolerance and allelic exclusion.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-B220 BioLegend Clone# RA3-6B2; RRID:AB_11203907 Anti-IgD BD Biosciences Clone# 11-26c-2a; RRID:AB_2738723 Anti-IgM eBioscience Clone# eB121-15F9; RRID:AB_10671539 Anti-IgM, polyclonal (for ELISA) SouthernBiotech Cat# 1021-01; RRID:AB_2687524 Anti-IgG1 (for ELISA) BioLegend or BD Biosciences Clone# A85-3; RRID:AB_394863 Anti-IgG2a (for ELISA) BioLegend or BD Biosciences Clone# RMG2a-62; RRID:AB_2563481 Anti-IgG2b (for ELISA) BioLegend or BD Biosciences Clone# RMG2b-1; RRID:AB_315065 Anti-IgG3 (for ELISA) BioLegend or BD Biosciences Clone# R2-38; RRID:AB_394841 Anti-CD43 BD Biosciences Clone# 1G10; RRID:AB_394748 Anti-CD21 BioLegend Clone# 7E9; RRID:AB_1953277 Anti-CD23 BD Biosciences Clone# B3B4; RRID:AB_2738526 Anti-CD24 BioLegend Clone# M1/69; RRID:AB_1595596 Anti-CD19 BD Biosciences Clone# 1D3; RRID:AB_2737915 Anti-Iglambda BioLegend Clone# RML-42; RRID:AB_961363 Anti-Iglambda SouthernBiotech Cat# 1060-02; RRID:AB_2794390 Anti-Igkappa BD Biosciences Clone# 187.1; RRID:AB_1727535 Anti-IL-7R BioLegend Clone# A7R34; RRID:AB_1937252 Anti-BAFFR R&D Systems Clone# BAF1357; RRID:AB_2271820 Anti-CXCR4 BioLegend Clone# L276F12; RRID:AB_2562785 Anti-CD93 eBioscience Clone# AA4.1; RID:AB_469466 Anti-CD1d BioLegend Clone# 1B1; RRID:AB_10683167 Anti-pH2AX BioLegend Clone# 2F3; RRID:AB_528919 Anti-pAKT (T308) Cell Signaling Technology Clone# C31E5E; RRID:AB_2255933 Anti-Foxo1 Cell Signaling Technology Clone# C29H4; RRID:AB_2106495 Anti-tri-methyl-histone H3 Cell Signaling Technology Clone# C36B11; RRID:AB_2799182 Anti-rabbit IgG-FITC SouthernBiotech Cat# 4050-02; RRID:AB_2795952 Anti-rabbit IgG-DyLight 649 BioLegend Cat# 406406; RRID:AB_1575135 Anti-3-83Ig(H + L) Produced in-house, University of Colorado Anschutz Medical Campus Clone# 54.1 Anti-B220 Microbeads Miltenyi Biotec Cat# 130-049-501 Anti-FITC Microbeads Miltenyi Biotec Cat# 130-058-701 Chemicals, peptides, and recombinant proteins Phosphate substrate Millipore Sigma #P5744 Streptavidin-AP SouthernBiotech #7105-05 Streptavidin-APC-Cy7 BD Biosciences #554063 BSA Fisher Scientific #BP1600-100 Critical commercial assays Zombie UV Fixable Viability Kit BioLegend #423108 eBioscienceTM Foxp3/Transcription Factor Staining Buffer Set eBioscience #00-5523-00 BioLegend Fixation Buffer BioLegend #420801 APC BrdU Flow Kit BD Pharmingen #552598 RNeasy PlusMini Kit Qiagen #74134 SuperScript IV VILO Master Mix Thermo Fisher Scientific #11756050 (Continued on next page) 16 Cell Reports 43, 114283, June 25, 2024 .. REAGENT or RESOURCE SOURCE IDENTIFIER Taqman Gene Expression Assay, Rag1, Mm01270936_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Rag2, Mm00501300_m1 Thermo Fisher Scientific #4448892 Taqman Gene Expression Assay, Foxo1, Mm00490671_m1 Thermo Fisher Scientific #4331182 Taqman Gene Expression Assay, Cxcr4, Mm01996749_s1 Thermo Fisher Scientific #4453320 Taqman Gene Expression Assay, Cd79b, Mm00434143_m1 Thermo Fisher Scientific #4331182 Taqman Universal PCR Master Mix Applied Biosystems #4364338 Experimental models: Organisms/strains 3-83Igi,H-2d mice Braun et al74 Jackson Laboratories #011074 3-83Igi,H-2b mice Halverson et al26 N/A 3-83Igi,Rosa26-P110*,mb1-Cre mice Greaves et al31 N/A Rosa26-P110* mice Srinivasan et al55 Jackson Laboratories #012343 Mb1-Cre mice Hobeika et al75 Jackson Laboratories #029412 3-83Igi,Foxo1fl/fl,mb1-Cre mice This paper N/A Foxo1fl/fl mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-Foxo1-AAA,mb1-Cre mice This paper N/A Foxo1-AAA mice Stephen Hedrick (UCSD) N/A 3-83Igi,Rosa26-P110*/Foxo1-AAA,mb1-Cre mice This paper N/A Software and algorithms Flowjo v10.9 BD https://www.flowjo.com/ GraphPad Prism 10 GraphPad https://www.graphpad.com/ QuantStudio 7 Flex Real-Time PCR System software Thermo Fisher Scientific https://www.thermofisher.com/ Other Fetal Bovine Serum Gemini #100106, Lot#050A07H00K
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