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lambda phosphatase  (Addgene inc)


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    Structured Review

    Addgene inc lambda phosphatase
    Lambda Phosphatase, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 24 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 24 article reviews
    lambda phosphatase - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Phosphorylation patterns modulate the transient secondary structure of RNA polymerase II CTD without altering its global conformation
    Article Snippet: .. BL21 (DE3) cells were co-transformed with plamids containing the kinase domain of human Dyrk1a (Addgene Plasmid #79690) and the lambda phosphatase (Addgene Plasmid #79748)( ). ..

    Article Title: Phosphorylation modulates secondary structure of intrinsically disorder regions in RNA polymerase II.
    Article Snippet: .. BL21 (DE3) cells were cotransformed with plasmids containing the kinase domain of human Dyrk1a (Addgene Plasmid #79690) and the lambda phosphatase (Addgene Plasmid #79748) (55). .. BL21 (DE3) cells were cotransformed with plasmids containing the kinase domain of human Dyrk1a (Addgene Plasmid #79690) and the lambda phosphatase (Addgene Plasmid #79748) (55).

    Article Title: Phosphorylation modulates secondary structure of intrinsically disorder regions in RNA polymerase II
    Article Snippet: .. BL21 (DE3) cells were cotransformed with plasmids containing the kinase domain of human Dyrk1a (Addgene Plasmid #79690) and the lambda phosphatase (Addgene Plasmid #79748) ( ). ..

    Article Title: Multivalent DNA and nucleosome acidic patch interactions specify VRK1 mitotic localization and activity
    Article Snippet: .. BL21(DE3)pLysS cells were first transformed with a plasmid encoding the Lambda phosphatase acquired as a gift from John Chodera, Nicholas Levinson and Markus Seeliger (Addgene plasmid #79748) and plated under spectinomycin and chloramphenicol selection. ..

    Expressing:

    Article Title: An atlas of substrate specificities for the human serine/threonine kinome
    Article Snippet: .. Bacterial expression constructs for DMPK1, CAMK1G, CAMK2G, PHKG2, CDKL1, GAK and lambda phosphatase were purchased from Addgene (Addgene, 1000000094) . .. Transformations were performed using BL21 Star cells (Thermo Fisher Scientific) unless specified otherwise.

    Article Title: A global atlas of substrate specificities for the human serine/threonine kinome
    Article Snippet: .. Bacterial expression constructs for DMPK1, CAMK1A, CAMK1G, CAMK2G, PHKG2, CDKL1, GAK, and lambda phosphatase were purchased from Addgene (Addgene Kit #1000000094) . .. Transformations were performed with BL21 Star cells (Thermo Fisher) unless specified otherwise.

    Construct:

    Article Title: An atlas of substrate specificities for the human serine/threonine kinome
    Article Snippet: .. Bacterial expression constructs for DMPK1, CAMK1G, CAMK2G, PHKG2, CDKL1, GAK and lambda phosphatase were purchased from Addgene (Addgene, 1000000094) . .. Transformations were performed using BL21 Star cells (Thermo Fisher Scientific) unless specified otherwise.

    Article Title: A global atlas of substrate specificities for the human serine/threonine kinome
    Article Snippet: .. Bacterial expression constructs for DMPK1, CAMK1A, CAMK1G, CAMK2G, PHKG2, CDKL1, GAK, and lambda phosphatase were purchased from Addgene (Addgene Kit #1000000094) . .. Transformations were performed with BL21 Star cells (Thermo Fisher) unless specified otherwise.

    other:

    Article Title: Phosphorylation modulates secondary structure of intrinsically disorder regions in RNA polymerase II.
    Article Snippet: pET His6 MBP TEV LIC cloning vector (1M) was a gift from Scott Gradia (Addgene plasmid # 29656; http://n2t.net/addgene:29656; RRID:Addgene_29656).

    Recombinant:

    Article Title: CaMKII monomers are sufficient for GluN2B binding, co-condensation, and synaptic potentiation.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: Cell lines Primary rat hippocampal cultures Laboratory of K. Ulrich Bayer N/A HEK293T cells ATCC CRL-3216 Experimental models: Organisms/strains GluN2B LA/RQ mice (C57BL/6J) Laboratory of Johannes Hell, Halt et al.9 N/A Recombinant DNA PSD95-FingR-GFP Laboratory of Don Arnold, Gross et al.47 Addgene #46295 pMH-Halo tag (‘‘HaloFill’’) Michael Huen Addgene #154144 CaMKIIα shRNA Laboratory of K. Ulrich Bayer, Barcomb et al.46 N/A pNES-iRFP-C1 Goulden et al.63 pNES-iRFP-C1, Addgene #116863 pmTurquoise2-T2A-Venus(L68V) Goedhart et al.32 pmTurquoise2-T2A-Venus(L68V), Addgene #60494 GFP-C1-CaMKIIα Shen et al.64 GFP-C1-CAMKIIalpha, Addgene #21226 Cry2olig Taslimi et al.34 N/A CIBN-mCherry-CaMKIIα Taslimi et al.34 N/A CMV-pDisplay-Halo Tobias et al.65 CMV-pDisplay-Halo, Addgene #223497 Signal-mCherry-TM(pDisplay)-GluN2B Made in house N/A mScarlet GluN2B (soluble) Made in house, Rumian et al.7 N/A GST-GluN2Bc Made in house, Bayer et al.8 N/A 6xHis-SUMO-CaMKIIα (6HS-CaMKII) Ozden et al.16 N/A λ phosphatase Albanese et al.66 Lambda Phosphatase, Addgene #79748 Software and algorithms Prism 10.5.0 Graphpad RRID: SCR_002798 FIJI ImageJ NIH RRID: SCR_003070 Slidebook 6.0 Intelligent Imaging Innovations (3i) RRID: SCR_014300 PyMOL Schrödinger RRID: SCR_000305 PyCorrFit 1.1.7 Lab of T. Weidemann, Müller et al.67 N/A Other HisPrep FF Nickel Sepharose column Cytiva 28936551 ll OPEN ACCESS Article e2 Molecular Cell 86, 1–14.e1–e6, October 1, 2026 .. All DNA constructs were either purchased from AddGene or received as a gift and were further cloned to include the indicated fluorophores, mutations, and truncations.

    shRNA:

    Article Title: CaMKII monomers are sufficient for GluN2B binding, co-condensation, and synaptic potentiation.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: Cell lines Primary rat hippocampal cultures Laboratory of K. Ulrich Bayer N/A HEK293T cells ATCC CRL-3216 Experimental models: Organisms/strains GluN2B LA/RQ mice (C57BL/6J) Laboratory of Johannes Hell, Halt et al.9 N/A Recombinant DNA PSD95-FingR-GFP Laboratory of Don Arnold, Gross et al.47 Addgene #46295 pMH-Halo tag (‘‘HaloFill’’) Michael Huen Addgene #154144 CaMKIIα shRNA Laboratory of K. Ulrich Bayer, Barcomb et al.46 N/A pNES-iRFP-C1 Goulden et al.63 pNES-iRFP-C1, Addgene #116863 pmTurquoise2-T2A-Venus(L68V) Goedhart et al.32 pmTurquoise2-T2A-Venus(L68V), Addgene #60494 GFP-C1-CaMKIIα Shen et al.64 GFP-C1-CAMKIIalpha, Addgene #21226 Cry2olig Taslimi et al.34 N/A CIBN-mCherry-CaMKIIα Taslimi et al.34 N/A CMV-pDisplay-Halo Tobias et al.65 CMV-pDisplay-Halo, Addgene #223497 Signal-mCherry-TM(pDisplay)-GluN2B Made in house N/A mScarlet GluN2B (soluble) Made in house, Rumian et al.7 N/A GST-GluN2Bc Made in house, Bayer et al.8 N/A 6xHis-SUMO-CaMKIIα (6HS-CaMKII) Ozden et al.16 N/A λ phosphatase Albanese et al.66 Lambda Phosphatase, Addgene #79748 Software and algorithms Prism 10.5.0 Graphpad RRID: SCR_002798 FIJI ImageJ NIH RRID: SCR_003070 Slidebook 6.0 Intelligent Imaging Innovations (3i) RRID: SCR_014300 PyMOL Schrödinger RRID: SCR_000305 PyCorrFit 1.1.7 Lab of T. Weidemann, Müller et al.67 N/A Other HisPrep FF Nickel Sepharose column Cytiva 28936551 ll OPEN ACCESS Article e2 Molecular Cell 86, 1–14.e1–e6, October 1, 2026 .. All DNA constructs were either purchased from AddGene or received as a gift and were further cloned to include the indicated fluorophores, mutations, and truncations.

    Software:

    Article Title: CaMKII monomers are sufficient for GluN2B binding, co-condensation, and synaptic potentiation.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Experimental models: Cell lines Primary rat hippocampal cultures Laboratory of K. Ulrich Bayer N/A HEK293T cells ATCC CRL-3216 Experimental models: Organisms/strains GluN2B LA/RQ mice (C57BL/6J) Laboratory of Johannes Hell, Halt et al.9 N/A Recombinant DNA PSD95-FingR-GFP Laboratory of Don Arnold, Gross et al.47 Addgene #46295 pMH-Halo tag (‘‘HaloFill’’) Michael Huen Addgene #154144 CaMKIIα shRNA Laboratory of K. Ulrich Bayer, Barcomb et al.46 N/A pNES-iRFP-C1 Goulden et al.63 pNES-iRFP-C1, Addgene #116863 pmTurquoise2-T2A-Venus(L68V) Goedhart et al.32 pmTurquoise2-T2A-Venus(L68V), Addgene #60494 GFP-C1-CaMKIIα Shen et al.64 GFP-C1-CAMKIIalpha, Addgene #21226 Cry2olig Taslimi et al.34 N/A CIBN-mCherry-CaMKIIα Taslimi et al.34 N/A CMV-pDisplay-Halo Tobias et al.65 CMV-pDisplay-Halo, Addgene #223497 Signal-mCherry-TM(pDisplay)-GluN2B Made in house N/A mScarlet GluN2B (soluble) Made in house, Rumian et al.7 N/A GST-GluN2Bc Made in house, Bayer et al.8 N/A 6xHis-SUMO-CaMKIIα (6HS-CaMKII) Ozden et al.16 N/A λ phosphatase Albanese et al.66 Lambda Phosphatase, Addgene #79748 Software and algorithms Prism 10.5.0 Graphpad RRID: SCR_002798 FIJI ImageJ NIH RRID: SCR_003070 Slidebook 6.0 Intelligent Imaging Innovations (3i) RRID: SCR_014300 PyMOL Schrödinger RRID: SCR_000305 PyCorrFit 1.1.7 Lab of T. Weidemann, Müller et al.67 N/A Other HisPrep FF Nickel Sepharose column Cytiva 28936551 ll OPEN ACCESS Article e2 Molecular Cell 86, 1–14.e1–e6, October 1, 2026 .. All DNA constructs were either purchased from AddGene or received as a gift and were further cloned to include the indicated fluorophores, mutations, and truncations.

    Transformation Assay:

    Article Title: Multivalent DNA and nucleosome acidic patch interactions specify VRK1 mitotic localization and activity
    Article Snippet: .. BL21(DE3)pLysS cells were first transformed with a plasmid encoding the Lambda phosphatase acquired as a gift from John Chodera, Nicholas Levinson and Markus Seeliger (Addgene plasmid #79748) and plated under spectinomycin and chloramphenicol selection. ..

    Selection:

    Article Title: Multivalent DNA and nucleosome acidic patch interactions specify VRK1 mitotic localization and activity
    Article Snippet: .. BL21(DE3)pLysS cells were first transformed with a plasmid encoding the Lambda phosphatase acquired as a gift from John Chodera, Nicholas Levinson and Markus Seeliger (Addgene plasmid #79748) and plated under spectinomycin and chloramphenicol selection. ..



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    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.
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    Image Search Results


    (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.

    Journal: PLOS One

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

    doi: 10.1371/journal.pone.0349130

    Figure Lengend 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.

    Article Snippet: DNA-PK inhibitor NU7441 was purchased from Tocris Bioscience. λ protein phosphatase (LPP) was purchased from New England Biolabs.

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