Review



λ protein phosphatase lpp  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 99
      Buy from Supplier

    Structured Review

    New England Biolabs λ protein phosphatase lpp
    (A) After standard ARE pulldowns, the AR-DNA-bound complexes were washed and resuspended in 1x kinase buffer with/without ATP at 30 o C for listed times. Gel mobility shifts in AR were detected as a function of ATP addition. All pulldowns had 100 nM R1881 present. Proteins were analyzed by immunoblotting. Lanes 3 and 4 represent the standard AR-ARE pulldown. (B) To test if the above AR gel mobility shifts were reflective of phosphorylation events, the AR-CoR complexes were treated with ATP or AMP-PNP (non-hydrolyzable analogue) for 30 min at 30 o C. The DNA-PK inhibitor NU7441 or <t>λ</t> <t>protein</t> <t>phosphatase</t> <t>(LPP)</t> were pre-incubated for 5 min before adding ATP. LPP buffer was used for the LPP control reaction. (C) DNA-PK plays a role in phosphorylating AR. ATP treatment of the AR-CoR complexes using HeLa NE where DNA-PKcs was immunodepleted reveals a reduction of AR gel mobility shifts ( left panel) with the loss of DNA-PK. Proteins were assayed by immunoblotting and the level of unphosphorylated AR was quantified by Image J after normalization to the loading control HDAC1 ( right panel). P-value here = 0.00011. (D) DNA-bound AR or ARv7 was incubated with purified heterotrimeric DNA-PK (i.e., DNA-PKcs and Ku70/80) and then treated (-/+) ATP (-/+) NU7441 to demonstrate direct phosphorylation of AR/ARv7 by DNA-PK. Gel mobility shifts were assayed by immunoblotting. (E) ATP treatment (-/+) NU7441 pre-treatment demonstrates that DNA-PK enzymatic activity is needed for stabilizing CAND1 and ANP32A in the AR/ARv7 complexes. Auto-phosphorylation of DNA-PKcs at serine 2056 (pS2056) is a marker of DNA-PK activity and its inhibition by NU7441. Proteins were assayed by immunoblotting. ARv7 was detected here with the N-20 antibody. (F) HeLa NE used in 3xARE-E4 cell-free transcription assays was pre-treated (-/+) NU7441 to determine effects on AR and ARv7 transcriptional activity. E4 mRNA transcript levels were measured by RT-qPCR. In panels A-C and E, HDAC1 was used as a loading control and ARE pulldowns were conducted using HeLa NE as listed. All AR reactions had 100 nM R1881 present. Short exp., Medium exp., and Long exp. denote short, medium, and long exposures during x-ray film development. P-values: for ARv7, p = 0.00015; for AR, p = 0.000035. **** p < 0.0005, ***** p < 0.0001.
    λ Protein Phosphatase Lpp, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2929 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/pmc13175348-42-8-15
    Average 99 stars, based on 2929 article reviews
    λ protein phosphatase lpp - by Bioz Stars, 2026-11
    99/100 stars

    Images

    1) Product Images from "Identification of CAND1 as a DNA-dependent protein kinase-regulated coactivator of androgen receptor and the ARv7 splice variant"

    Article Title: Identification of CAND1 as a DNA-dependent protein kinase-regulated coactivator of androgen receptor and the ARv7 splice variant

    Journal: PLOS One

    doi: 10.1371/journal.pone.0349130

    (A) After standard ARE pulldowns, the AR-DNA-bound complexes were washed and resuspended in 1x kinase buffer with/without ATP at 30 o C for listed times. Gel mobility shifts in AR were detected as a function of ATP addition. All pulldowns had 100 nM R1881 present. Proteins were analyzed by immunoblotting. Lanes 3 and 4 represent the standard AR-ARE pulldown. (B) To test if the above AR gel mobility shifts were reflective of phosphorylation events, the AR-CoR complexes were treated with ATP or AMP-PNP (non-hydrolyzable analogue) for 30 min at 30 o C. The DNA-PK inhibitor NU7441 or λ protein phosphatase (LPP) were pre-incubated for 5 min before adding ATP. LPP buffer was used for the LPP control reaction. (C) DNA-PK plays a role in phosphorylating AR. ATP treatment of the AR-CoR complexes using HeLa NE where DNA-PKcs was immunodepleted reveals a reduction of AR gel mobility shifts ( left panel) with the loss of DNA-PK. Proteins were assayed by immunoblotting and the level of unphosphorylated AR was quantified by Image J after normalization to the loading control HDAC1 ( right panel). P-value here = 0.00011. (D) DNA-bound AR or ARv7 was incubated with purified heterotrimeric DNA-PK (i.e., DNA-PKcs and Ku70/80) and then treated (-/+) ATP (-/+) NU7441 to demonstrate direct phosphorylation of AR/ARv7 by DNA-PK. Gel mobility shifts were assayed by immunoblotting. (E) ATP treatment (-/+) NU7441 pre-treatment demonstrates that DNA-PK enzymatic activity is needed for stabilizing CAND1 and ANP32A in the AR/ARv7 complexes. Auto-phosphorylation of DNA-PKcs at serine 2056 (pS2056) is a marker of DNA-PK activity and its inhibition by NU7441. Proteins were assayed by immunoblotting. ARv7 was detected here with the N-20 antibody. (F) HeLa NE used in 3xARE-E4 cell-free transcription assays was pre-treated (-/+) NU7441 to determine effects on AR and ARv7 transcriptional activity. E4 mRNA transcript levels were measured by RT-qPCR. In panels A-C and E, HDAC1 was used as a loading control and ARE pulldowns were conducted using HeLa NE as listed. All AR reactions had 100 nM R1881 present. Short exp., Medium exp., and Long exp. denote short, medium, and long exposures during x-ray film development. P-values: for ARv7, p = 0.00015; for AR, p = 0.000035. **** p < 0.0005, ***** p < 0.0001.
    Figure Legend Snippet: (A) After standard ARE pulldowns, the AR-DNA-bound complexes were washed and resuspended in 1x kinase buffer with/without ATP at 30 o C for listed times. Gel mobility shifts in AR were detected as a function of ATP addition. All pulldowns had 100 nM R1881 present. Proteins were analyzed by immunoblotting. Lanes 3 and 4 represent the standard AR-ARE pulldown. (B) To test if the above AR gel mobility shifts were reflective of phosphorylation events, the AR-CoR complexes were treated with ATP or AMP-PNP (non-hydrolyzable analogue) for 30 min at 30 o C. The DNA-PK inhibitor NU7441 or λ protein phosphatase (LPP) were pre-incubated for 5 min before adding ATP. LPP buffer was used for the LPP control reaction. (C) DNA-PK plays a role in phosphorylating AR. ATP treatment of the AR-CoR complexes using HeLa NE where DNA-PKcs was immunodepleted reveals a reduction of AR gel mobility shifts ( left panel) with the loss of DNA-PK. Proteins were assayed by immunoblotting and the level of unphosphorylated AR was quantified by Image J after normalization to the loading control HDAC1 ( right panel). P-value here = 0.00011. (D) DNA-bound AR or ARv7 was incubated with purified heterotrimeric DNA-PK (i.e., DNA-PKcs and Ku70/80) and then treated (-/+) ATP (-/+) NU7441 to demonstrate direct phosphorylation of AR/ARv7 by DNA-PK. Gel mobility shifts were assayed by immunoblotting. (E) ATP treatment (-/+) NU7441 pre-treatment demonstrates that DNA-PK enzymatic activity is needed for stabilizing CAND1 and ANP32A in the AR/ARv7 complexes. Auto-phosphorylation of DNA-PKcs at serine 2056 (pS2056) is a marker of DNA-PK activity and its inhibition by NU7441. Proteins were assayed by immunoblotting. ARv7 was detected here with the N-20 antibody. (F) HeLa NE used in 3xARE-E4 cell-free transcription assays was pre-treated (-/+) NU7441 to determine effects on AR and ARv7 transcriptional activity. E4 mRNA transcript levels were measured by RT-qPCR. In panels A-C and E, HDAC1 was used as a loading control and ARE pulldowns were conducted using HeLa NE as listed. All AR reactions had 100 nM R1881 present. Short exp., Medium exp., and Long exp. denote short, medium, and long exposures during x-ray film development. P-values: for ARv7, p = 0.00015; for AR, p = 0.000035. **** p < 0.0005, ***** p < 0.0001.

    Techniques Used: Western Blot, Phospho-proteomics, Incubation, Control, Purification, Activity Assay, Marker, Inhibition, Quantitative RT-PCR

    Related Articles

    Incubation:

    Article Title: AKT Phosphorylation of FOXK2 at Ser398 Impacts Breast Cancer Cell Growth and Viability
    Article Snippet: .. Afterwards, 100 μg of protein were incubated in a total of 50 μL containing 400 units of lambda protein phosphatase (λPP; P0753S, New England BioLabs Inc., Ipswich, MA, USA), 1× PMP (NEBuffer Pack for Protein MetalloPhosphatases; B0761S, New England BioLabs Inc.) and 1 mM MnCl2 (B1761S, New England BioLabs Inc.) for 1 h at 37°C. ..

    Article Title: Single-molecule resolution of post-translational modification on GRK4 with cyclodextrin/SiN x hybrid nanopore.
    Article Snippet: .. A 100 μL of the mixture containing 10 μM of pep-T182-P and 200 U of lambda protein phosphatase in 1 mM MnCl2 and NEbuffer for PMP was incubated at 30 ◦C for 2 h, a control assay is under identical condition but without lambda protein phosphatase. ..

    Article Title: Identification of NEK9 as a novel substrate of protein kinase B.
    Article Snippet: Protein kinase B (PKB/Akt) regulates diverse cellular processes through phosphorylation of numerous substrate proteins downstream of phosphatidylinositol 3-kinase (PI3K).. Although many PKB substrates have been identified, additional substrates remain to be characterized.. NEK9 is an essential serine/threonine kinase involved in mitotic progression and contains two putative PKB consensus phosphorylation motifs.

    Article Title: The PvAURK-PvGRFs regulatory module controls salt tolerance in common bean (Phaseolus vulgaris L.).
    Article Snippet: .. For the dephosphorylation assay, the extracted protein samples were incubated with λ-protein phosphatase (λ-PPase, New England Biolabs) at 30 ◦C for 45 min according to the manufacturer's instructions. ..

    Article Title: VASH1 phosphorylation regulates tubulin detyrosination via CNP/PRKG1α signaling in DRG neurons
    Article Snippet: .. For phosphatase treatment experiments, bead-bound complexes were incubated with phosphatase (Lambda protein phosphatase, New England Biolabs) for 30 min at 30°C according to the manufacturer’s protocol before elution. ..

    Control:

    Article Title: Single-molecule resolution of post-translational modification on GRK4 with cyclodextrin/SiN x hybrid nanopore.
    Article Snippet: .. A 100 μL of the mixture containing 10 μM of pep-T182-P and 200 U of lambda protein phosphatase in 1 mM MnCl2 and NEbuffer for PMP was incubated at 30 ◦C for 2 h, a control assay is under identical condition but without lambda protein phosphatase. ..

    Thin Layer Chromatography:

    Article Title: A bifunctional ARHGEF17-AS1 locus encodes MSEP, a protein that supports mitotic spindle organization.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-α-tubulin (clone DM1A) Sigma-Aldrich Cat# T6199; RRID: AB_477583 Rabbit polyclonal anti-α-tubulin Proteintech Cat# 11224-1-AP; RRID: AB_2210206 Mouse monoclonal anti-γ-tubulin (clone GTU-88) Sigma-Aldrich Cat# T5326; RRID: AB_532292 Rabbit polyclonal anti-γ-tubulin Sigma-Aldrich Cat# T5192; RRID: AB_261690 Mouse monoclonal anti-RPS3 (clone 2G7H4) Proteintech Cat# 66046-1-Ig; RRID: AB_11182493 Rabbit polyclonal anti-RPS3 Novus Biologicals Cat# NBP1-33691; RRID: AB_2180759 Rabbit monoclonal anti-RPS3 (clone D50G7) Cell Signaling Technology Cat# 9538S; RRID: AB_10622028 Rabbit polyclonal anti-RNF10 Proteintech Cat# 16936-1-AP; RRID: AB_2181846 Mouse monoclonal anti-TPX2 (clone 2E10C1) Proteintech Cat# 68712-1-Ig; RRID: AB_3670405 Rabbit polyclonal anti-TPX2 Proteintech Cat# 11741-1-AP; RRID: AB_2208895 Mouse monoclonal anti-KPNA2 (clone 2G9C1) Proteintech Cat# 66870-1-Ig; RRID: AB_2882207 Rabbit polyclonal anti-KPNA2 Proteintech Cat# 10819-1-AP; RRID: AB_2265526 Rabbit polyclonal anti-DYNC1H1/dynein heavy chain Proteintech Cat# 12345-1-AP; RRID: AB_2261765 Mouse monoclonal anti-cytoplasmic dynein intermediate chain (clone 74.1) Abcam Cat# ab23905; RRID: AB_2096669 Mouse monoclonal anti-FLAG M2 (clone M2) Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse monoclonal anti-GFP tag (clone 1E10H7) Proteintech Cat# 66002-1-Ig; RRID: AB_11182611 Rabbit polyclonal anti-GFP Abcam Cat# ab290; RRID: AB_303395 Rabbit monoclonal anti-GAPDH (clone D16H11) Cell Signaling Technology Cat# 5174; RRID: AB_10622025 Rabbit monoclonal anti-Aurora-A/AIK (clone 1G4) Cell Signaling Technology Cat# 4718; RRID: AB_2061482 Mouse monoclonal anti-Aurora-A (clone 1F4B10) Proteintech Cat# 66757-1-Ig; RRID: AB_2882103 Rabbit monoclonal anti-WEE1 (clone D10D2) Cell Signaling Technology Cat# 13084; RRID: AB_2713924 Rabbit monoclonal anti-PLK1 (clone 208G4) Cell Signaling Technology Cat# 4513; RRID: AB_2167409 Rabbit polyclonal anti-vinculin Proteintech Cat# 26520-1-AP; RRID: AB_2868558 Normal mouse IgG MilliporeSigma Cat# 12–371; RRID: AB_145840 Normal rabbit IgG MilliporeSigma Cat# 12–370; RRID: AB_145841 Rabbit polyclonal anti-MSEP; immunogen CGHRPQRLHPGRP (residues 257–269) This paper; Innovagen AB N/A Anti-MYC tag antibody Abcam Cat# ab32; RRID: AB_303599 Anti-cyclin B1 antibody Abcam Cat# ab32053; RRID: AB_731779 Anti-total PARP antibody Abcam Cat# ab191217; RRID: AB_2861274 (Continued on next page) 16 Cell Reports 45, 118026, October 27, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER N-Acetyl-D-sphingosine; C2 ceramide (d18:1/2:0), ≥97% (TLC) Sigma-Aldrich Cat# A7191; CAS: 3102–57-6 Lambda Protein Phosphatase New England Biolabs Cat# P0753S Puromycin dihydrochloride Sigma-Aldrich Cat# P8833; CAS: 58–58-2 Hygromycin B Sigma-Aldrich Cat# H3274; CAS: 31282–04-9 TRIzol Reagent Invitrogen/Thermo Fisher Scientific Cat# 15596026 ProLong Gold Antifade Mountant Invitrogen/Thermo Fisher Scientific Cat# P36930 RNase A Invitrogen/Thermo Fisher Scientific Cat# 12091021 Halt Protease and Phosphatase Inhibitor Cocktail Thermo Fisher Scientific Cat# 78440 Phosphatase Inhibitor Cocktail Set IV MilliporeSigma Cat# 524628 Protein G magnetic beads Invitrogen/Thermo Fisher Scientific Cat# 10004D MSEP immunogen peptide CGHRPQRLHPGRP (residues 257–269) This paper; Innovagen AB N/A Critical commercial assays Lipofectamine RNAiMAX Transfection Reagent Invitrogen/Thermo Fisher Scientific Cat# 13778150 Lipofectamine 2000 Transfection Reagent Invitrogen/Thermo Fisher Scientific Cat# 11668019 SF Cell Line 4D-Nucleofector X Kit L Lonza Cat# V4XC-2012 ReliaPrep RNA Cell Miniprep System Promega Cat# Z6010, Z6011. .. ImProm-II Reverse Transcription System Promega Cat# A3800 Power SYBR Green PCR Master Mix Applied Biosystems/Thermo Fisher Scientific Cat# 4367659 RNAscope Multiplex Fluorescent Reagent Kit v2 Advanced Cell Diagnostics/Bio-Techne Cat# 323100 Duolink In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Pierce BCA Protein Assay Kit Thermo Fisher Scientific Cat# 23225 NuPAGE Novex 4–12% Bis-Tris protein gels Invitrogen/Thermo Fisher Scientific Cat# NP0321 Microcon 30 kDa centrifugal filter units Merck Millipore Cat# MRCF0R030 CellTiter-Glo 2.0 Cell Viability Assay Promega Cat# G9241 Deposited data RNA-sequencing data reported in this study This paper; NCBI Gene Expression Omnibus GEO: GSE274156; https://www.ncbi.nlm.

    Magnetic Beads:

    Article Title: A bifunctional ARHGEF17-AS1 locus encodes MSEP, a protein that supports mitotic spindle organization.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-α-tubulin (clone DM1A) Sigma-Aldrich Cat# T6199; RRID: AB_477583 Rabbit polyclonal anti-α-tubulin Proteintech Cat# 11224-1-AP; RRID: AB_2210206 Mouse monoclonal anti-γ-tubulin (clone GTU-88) Sigma-Aldrich Cat# T5326; RRID: AB_532292 Rabbit polyclonal anti-γ-tubulin Sigma-Aldrich Cat# T5192; RRID: AB_261690 Mouse monoclonal anti-RPS3 (clone 2G7H4) Proteintech Cat# 66046-1-Ig; RRID: AB_11182493 Rabbit polyclonal anti-RPS3 Novus Biologicals Cat# NBP1-33691; RRID: AB_2180759 Rabbit monoclonal anti-RPS3 (clone D50G7) Cell Signaling Technology Cat# 9538S; RRID: AB_10622028 Rabbit polyclonal anti-RNF10 Proteintech Cat# 16936-1-AP; RRID: AB_2181846 Mouse monoclonal anti-TPX2 (clone 2E10C1) Proteintech Cat# 68712-1-Ig; RRID: AB_3670405 Rabbit polyclonal anti-TPX2 Proteintech Cat# 11741-1-AP; RRID: AB_2208895 Mouse monoclonal anti-KPNA2 (clone 2G9C1) Proteintech Cat# 66870-1-Ig; RRID: AB_2882207 Rabbit polyclonal anti-KPNA2 Proteintech Cat# 10819-1-AP; RRID: AB_2265526 Rabbit polyclonal anti-DYNC1H1/dynein heavy chain Proteintech Cat# 12345-1-AP; RRID: AB_2261765 Mouse monoclonal anti-cytoplasmic dynein intermediate chain (clone 74.1) Abcam Cat# ab23905; RRID: AB_2096669 Mouse monoclonal anti-FLAG M2 (clone M2) Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse monoclonal anti-GFP tag (clone 1E10H7) Proteintech Cat# 66002-1-Ig; RRID: AB_11182611 Rabbit polyclonal anti-GFP Abcam Cat# ab290; RRID: AB_303395 Rabbit monoclonal anti-GAPDH (clone D16H11) Cell Signaling Technology Cat# 5174; RRID: AB_10622025 Rabbit monoclonal anti-Aurora-A/AIK (clone 1G4) Cell Signaling Technology Cat# 4718; RRID: AB_2061482 Mouse monoclonal anti-Aurora-A (clone 1F4B10) Proteintech Cat# 66757-1-Ig; RRID: AB_2882103 Rabbit monoclonal anti-WEE1 (clone D10D2) Cell Signaling Technology Cat# 13084; RRID: AB_2713924 Rabbit monoclonal anti-PLK1 (clone 208G4) Cell Signaling Technology Cat# 4513; RRID: AB_2167409 Rabbit polyclonal anti-vinculin Proteintech Cat# 26520-1-AP; RRID: AB_2868558 Normal mouse IgG MilliporeSigma Cat# 12–371; RRID: AB_145840 Normal rabbit IgG MilliporeSigma Cat# 12–370; RRID: AB_145841 Rabbit polyclonal anti-MSEP; immunogen CGHRPQRLHPGRP (residues 257–269) This paper; Innovagen AB N/A Anti-MYC tag antibody Abcam Cat# ab32; RRID: AB_303599 Anti-cyclin B1 antibody Abcam Cat# ab32053; RRID: AB_731779 Anti-total PARP antibody Abcam Cat# ab191217; RRID: AB_2861274 (Continued on next page) 16 Cell Reports 45, 118026, October 27, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER N-Acetyl-D-sphingosine; C2 ceramide (d18:1/2:0), ≥97% (TLC) Sigma-Aldrich Cat# A7191; CAS: 3102–57-6 Lambda Protein Phosphatase New England Biolabs Cat# P0753S Puromycin dihydrochloride Sigma-Aldrich Cat# P8833; CAS: 58–58-2 Hygromycin B Sigma-Aldrich Cat# H3274; CAS: 31282–04-9 TRIzol Reagent Invitrogen/Thermo Fisher Scientific Cat# 15596026 ProLong Gold Antifade Mountant Invitrogen/Thermo Fisher Scientific Cat# P36930 RNase A Invitrogen/Thermo Fisher Scientific Cat# 12091021 Halt Protease and Phosphatase Inhibitor Cocktail Thermo Fisher Scientific Cat# 78440 Phosphatase Inhibitor Cocktail Set IV MilliporeSigma Cat# 524628 Protein G magnetic beads Invitrogen/Thermo Fisher Scientific Cat# 10004D MSEP immunogen peptide CGHRPQRLHPGRP (residues 257–269) This paper; Innovagen AB N/A Critical commercial assays Lipofectamine RNAiMAX Transfection Reagent Invitrogen/Thermo Fisher Scientific Cat# 13778150 Lipofectamine 2000 Transfection Reagent Invitrogen/Thermo Fisher Scientific Cat# 11668019 SF Cell Line 4D-Nucleofector X Kit L Lonza Cat# V4XC-2012 ReliaPrep RNA Cell Miniprep System Promega Cat# Z6010, Z6011. .. ImProm-II Reverse Transcription System Promega Cat# A3800 Power SYBR Green PCR Master Mix Applied Biosystems/Thermo Fisher Scientific Cat# 4367659 RNAscope Multiplex Fluorescent Reagent Kit v2 Advanced Cell Diagnostics/Bio-Techne Cat# 323100 Duolink In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Pierce BCA Protein Assay Kit Thermo Fisher Scientific Cat# 23225 NuPAGE Novex 4–12% Bis-Tris protein gels Invitrogen/Thermo Fisher Scientific Cat# NP0321 Microcon 30 kDa centrifugal filter units Merck Millipore Cat# MRCF0R030 CellTiter-Glo 2.0 Cell Viability Assay Promega Cat# G9241 Deposited data RNA-sequencing data reported in this study This paper; NCBI Gene Expression Omnibus GEO: GSE274156; https://www.ncbi.nlm.

    Transfection:

    Article Title: A bifunctional ARHGEF17-AS1 locus encodes MSEP, a protein that supports mitotic spindle organization.
    Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-α-tubulin (clone DM1A) Sigma-Aldrich Cat# T6199; RRID: AB_477583 Rabbit polyclonal anti-α-tubulin Proteintech Cat# 11224-1-AP; RRID: AB_2210206 Mouse monoclonal anti-γ-tubulin (clone GTU-88) Sigma-Aldrich Cat# T5326; RRID: AB_532292 Rabbit polyclonal anti-γ-tubulin Sigma-Aldrich Cat# T5192; RRID: AB_261690 Mouse monoclonal anti-RPS3 (clone 2G7H4) Proteintech Cat# 66046-1-Ig; RRID: AB_11182493 Rabbit polyclonal anti-RPS3 Novus Biologicals Cat# NBP1-33691; RRID: AB_2180759 Rabbit monoclonal anti-RPS3 (clone D50G7) Cell Signaling Technology Cat# 9538S; RRID: AB_10622028 Rabbit polyclonal anti-RNF10 Proteintech Cat# 16936-1-AP; RRID: AB_2181846 Mouse monoclonal anti-TPX2 (clone 2E10C1) Proteintech Cat# 68712-1-Ig; RRID: AB_3670405 Rabbit polyclonal anti-TPX2 Proteintech Cat# 11741-1-AP; RRID: AB_2208895 Mouse monoclonal anti-KPNA2 (clone 2G9C1) Proteintech Cat# 66870-1-Ig; RRID: AB_2882207 Rabbit polyclonal anti-KPNA2 Proteintech Cat# 10819-1-AP; RRID: AB_2265526 Rabbit polyclonal anti-DYNC1H1/dynein heavy chain Proteintech Cat# 12345-1-AP; RRID: AB_2261765 Mouse monoclonal anti-cytoplasmic dynein intermediate chain (clone 74.1) Abcam Cat# ab23905; RRID: AB_2096669 Mouse monoclonal anti-FLAG M2 (clone M2) Sigma-Aldrich Cat# F1804; RRID: AB_262044 Mouse monoclonal anti-GFP tag (clone 1E10H7) Proteintech Cat# 66002-1-Ig; RRID: AB_11182611 Rabbit polyclonal anti-GFP Abcam Cat# ab290; RRID: AB_303395 Rabbit monoclonal anti-GAPDH (clone D16H11) Cell Signaling Technology Cat# 5174; RRID: AB_10622025 Rabbit monoclonal anti-Aurora-A/AIK (clone 1G4) Cell Signaling Technology Cat# 4718; RRID: AB_2061482 Mouse monoclonal anti-Aurora-A (clone 1F4B10) Proteintech Cat# 66757-1-Ig; RRID: AB_2882103 Rabbit monoclonal anti-WEE1 (clone D10D2) Cell Signaling Technology Cat# 13084; RRID: AB_2713924 Rabbit monoclonal anti-PLK1 (clone 208G4) Cell Signaling Technology Cat# 4513; RRID: AB_2167409 Rabbit polyclonal anti-vinculin Proteintech Cat# 26520-1-AP; RRID: AB_2868558 Normal mouse IgG MilliporeSigma Cat# 12–371; RRID: AB_145840 Normal rabbit IgG MilliporeSigma Cat# 12–370; RRID: AB_145841 Rabbit polyclonal anti-MSEP; immunogen CGHRPQRLHPGRP (residues 257–269) This paper; Innovagen AB N/A Anti-MYC tag antibody Abcam Cat# ab32; RRID: AB_303599 Anti-cyclin B1 antibody Abcam Cat# ab32053; RRID: AB_731779 Anti-total PARP antibody Abcam Cat# ab191217; RRID: AB_2861274 (Continued on next page) 16 Cell Reports 45, 118026, October 27, 2026 .. REAGENT or RESOURCE SOURCE IDENTIFIER N-Acetyl-D-sphingosine; C2 ceramide (d18:1/2:0), ≥97% (TLC) Sigma-Aldrich Cat# A7191; CAS: 3102–57-6 Lambda Protein Phosphatase New England Biolabs Cat# P0753S Puromycin dihydrochloride Sigma-Aldrich Cat# P8833; CAS: 58–58-2 Hygromycin B Sigma-Aldrich Cat# H3274; CAS: 31282–04-9 TRIzol Reagent Invitrogen/Thermo Fisher Scientific Cat# 15596026 ProLong Gold Antifade Mountant Invitrogen/Thermo Fisher Scientific Cat# P36930 RNase A Invitrogen/Thermo Fisher Scientific Cat# 12091021 Halt Protease and Phosphatase Inhibitor Cocktail Thermo Fisher Scientific Cat# 78440 Phosphatase Inhibitor Cocktail Set IV MilliporeSigma Cat# 524628 Protein G magnetic beads Invitrogen/Thermo Fisher Scientific Cat# 10004D MSEP immunogen peptide CGHRPQRLHPGRP (residues 257–269) This paper; Innovagen AB N/A Critical commercial assays Lipofectamine RNAiMAX Transfection Reagent Invitrogen/Thermo Fisher Scientific Cat# 13778150 Lipofectamine 2000 Transfection Reagent Invitrogen/Thermo Fisher Scientific Cat# 11668019 SF Cell Line 4D-Nucleofector X Kit L Lonza Cat# V4XC-2012 ReliaPrep RNA Cell Miniprep System Promega Cat# Z6010, Z6011. .. ImProm-II Reverse Transcription System Promega Cat# A3800 Power SYBR Green PCR Master Mix Applied Biosystems/Thermo Fisher Scientific Cat# 4367659 RNAscope Multiplex Fluorescent Reagent Kit v2 Advanced Cell Diagnostics/Bio-Techne Cat# 323100 Duolink In Situ Red Starter Kit Mouse/Rabbit MilliporeSigma Cat# DUO92101 Pierce BCA Protein Assay Kit Thermo Fisher Scientific Cat# 23225 NuPAGE Novex 4–12% Bis-Tris protein gels Invitrogen/Thermo Fisher Scientific Cat# NP0321 Microcon 30 kDa centrifugal filter units Merck Millipore Cat# MRCF0R030 CellTiter-Glo 2.0 Cell Viability Assay Promega Cat# G9241 Deposited data RNA-sequencing data reported in this study This paper; NCBI Gene Expression Omnibus GEO: GSE274156; https://www.ncbi.nlm.

    De-Phosphorylation Assay:

    Article Title: The PvAURK-PvGRFs regulatory module controls salt tolerance in common bean (Phaseolus vulgaris L.).
    Article Snippet: .. For the dephosphorylation assay, the extracted protein samples were incubated with λ-protein phosphatase (λ-PPase, New England Biolabs) at 30 ◦C for 45 min according to the manufacturer's instructions. ..



    Similar Products

    phosphatase  (New England Biolabs)
    99
    New England Biolabs phosphatase
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    Phosphatase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/bio_rxiv__64898__2026__10__14__759141-247-9-13
    Average 99 stars, based on 1 article reviews
    phosphatase - by Bioz Stars, 2026-11
    99/100 stars
    		  Buy from Supplier
    lambda protein phosphatase  (New England Biolabs)
    99
    New England Biolabs lambda protein phosphatase
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    Lambda Protein Phosphatase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/10__1096_slash_fj__202600413rrr-99-17-22
    Average 99 stars, based on 1 article reviews
    lambda protein phosphatase - by Bioz Stars, 2026-11
    99/100 stars
    		  Buy from Supplier
    lambda phosphatase treatments  (New England Biolabs)
    99
    New England Biolabs lambda phosphatase treatments
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    Lambda Phosphatase Treatments, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/pm42847695-325-0-2
    Average 99 stars, based on 1 article reviews
    lambda phosphatase treatments - by Bioz Stars, 2026-11
    99/100 stars
    		  Buy from Supplier
    λ protein phosphatase  (New England Biolabs)
    99
    New England Biolabs λ protein phosphatase
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    λ Protein Phosphatase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/pm42837978-174-11-14
    Average 99 stars, based on 1 article reviews
    λ protein phosphatase - by Bioz Stars, 2026-11
    99/100 stars
    		  Buy from Supplier
    lambda phosphatase  (New England Biolabs)
    99
    New England Biolabs lambda phosphatase
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    Lambda Phosphatase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/pm42815464-353-32-34
    Average 99 stars, based on 1 article reviews
    lambda phosphatase - by Bioz Stars, 2026-11
    99/100 stars
    		  Buy from Supplier
    lambda pp  (New England Biolabs)
    99
    New England Biolabs lambda pp
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    Lambda Pp, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/pm42837813-62-23-27
    Average 99 stars, based on 1 article reviews
    lambda pp - by Bioz Stars, 2026-11
    99/100 stars
    		  Buy from Supplier
    lambda protein phosphatase new england biolabs  (New England Biolabs)
    99
    New England Biolabs lambda protein phosphatase new england biolabs
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    Lambda Protein Phosphatase New England Biolabs, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Protein+Phosphatase/pm42789387-519-16-19
    Average 99 stars, based on 1 article reviews
    lambda protein phosphatase new england biolabs - by Bioz Stars, 2026-11
    99/100 stars
    		  Buy from Supplier
    lambda phosphatase  (Addgene inc)
    94
    Addgene inc lambda phosphatase
    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with <t>Lambda</t> protein <t>phosphatase</t> results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.
    Lambda Phosphatase, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lambda+phosphatase/Lambda+Phosphatase+(Plasmid+%2379748)/pm42777704-252-140-142
    Average 94 stars, based on 1 article reviews
    lambda phosphatase - by Bioz Stars, 2026-11
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with Lambda protein phosphatase results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.

    Journal: bioRxiv

    Article Title: VASH1 phosphorylation regulates tubulin detyrosination via CNP/PRKG1α signaling in DRG neurons

    doi: 10.64898/2026.10.14.759141

    Figure Lengend Snippet: (A) Representative confocal images of axon with growth cones stained for tyrosinated α-tubulin (Tyr, red), F-actin (phalloidin, magenta), and phosphorylated-VASH1 (P[S337]VASH1, gray). Scale bar, 10 µm. ( B ) Quantification of P[S337]VASH1 in growth cones of WT and Vash1 KO neurons. n/N: 67/5 (WT), 111/7 (KO) with n indicating the number of growth cones and N of E13 embryos. Data are expressed as mean ± SD and as fold change over WT. ( C ) Quantification of P[S337]VASH1 in growth cones without treatment or with CNP treatment. n/N: 61/4 (control), 48/3 (CNP) with n indicating the number of growth cones and N the number of E13 embryos. Data are expressed as mean ± SD and as fold change over control. Statistical significance was determined using Mann-Witney test. **** p < 0.0001, ns p > 0.05. ( D, E ) Detection by immunoblot of VASH1 protein and its phosphorylated form in cultured DRG neurons after immunoprecipitation using an anti-SVBP antibody. β-tubulin staining shows similar protein content in the immunoprecipitation inputs. ( D ) Both VASH1 protein and its phosphorylated form are detected in WT but not in Vash1 KO DRGs. Treatment with Lambda protein phosphatase results in the loss of P[S337]VASH1 band. ( E, F ) CNP induces phosphorylation of S337 of VASH1, which is reversed by DT-3, as shown by representative immunoblots from an experiment done with cultured DRG explants ( E ) and quantifications of the P[S337]VASH1/VASH1 ratios from 4 independent experiments ( F ). In F , triangles correspond to data from immunoprecipitation experiments done with cultured DRG dissociated neurons, and circle to immunoprecipitation experiments done with cultured DRG explants.

    Article Snippet: For phosphatase treatment experiments, bead-bound complexes were incubated with phosphatase (Lambda protein phosphatase, New England Biolabs) for 30 min at 30°C according to the manufacturer’s protocol before elution.

    Techniques: Confocal, Staining, Control, Western Blot, Cell Culture, Immunoprecipitation, Phospho-proteomics