λ protein phosphatase lpp (New England Biolabs)
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λ protein phosphatase lpp
λ Protein Phosphatase Lpp, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2909 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/lpp/Lambda+Protein+Phosphatase/pmc13175348-42-8-15
Average 99 stars, based on 2909 article reviews
λ Protein Phosphatase Lpp, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2909 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/lpp/Lambda+Protein+Phosphatase/pmc13175348-42-8-15
Average 99 stars, based on 2909 article reviews
λ protein phosphatase lpp - by Bioz Stars,
2026-09
99/100 stars
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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
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
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Purification:Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units protein phosphatase (LPP) in a total reaction volume of 50 l following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units λ protein phosphatase (LPP) in a total reaction volume of 50 μl following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of In Vitro:Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units protein phosphatase (LPP) in a total reaction volume of 50 l following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units λ protein phosphatase (LPP) in a total reaction volume of 50 μl following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of Kinase Assay:Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units protein phosphatase (LPP) in a total reaction volume of 50 l following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units λ protein phosphatase (LPP) in a total reaction volume of 50 μl following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of Incubation:Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units protein phosphatase (LPP) in a total reaction volume of 50 l following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of Article Title: Endogenous IGFBP-3 Mediates Intrinsic Apoptosis Through Modulation of Nur77 Phosphorylation and Nuclear Export. Article Snippet: .. Lysates were incubated Article Title: Phosphorylation of FANCD2 Inhibits the FANCD2/FANCI Complex and Suppresses the Fanconi Anemia Pathway in the Absence of DNA Damage Article Snippet: In vitro Phosphorylation and Dephosphorylation Assays Phosphorylation reaction containing FANCD2/FANCI and CK2 (P6010, NEB) were incubated at 30 C for 30 min in 50 mM Tris (pH 7.5), 10 mM MgCl2, 2 mM ATP and 2 mM DTT. .. Dephosphorylation reaction containing FANCD2/FANCI and Article Title: Untangling the interactions: Structural basis of nitric oxide regulated cyclic-di-GMP metabolism in bacteria Article Snippet: .. 70 pL of the supernatant was mixed with 10 uL of 3U/ml Article Title: Ultrasonic Reporter of Kinase Activity Article Snippet: For all phosphorylation assays, 250 units of PKA (P6000L; NEB; specific activity: 1 pmol/min/unit), 1.5 μg of human CaMKIIα (Sino Biological; specific activity: 160 nmol/min/mg), or 0.1 μg of PKC alpha (Abcam; specific activity: 3,200 nmol/min/mg) per μL of engineered GVs at OD500 = 10 were mixed with 1 mM ATP (Sigma-Aldrich) in NEBufferTM for Protein Kinases (50 mM Tris-HCl, 10 mM MgCl 2 , 0.1 mM EDTA, 2 mM DTT, 0.01% Brij 35, pH 7.5; NEB) and incubated with slow rotation at 37°C for 10–12 hours, unless otherwise specified. .. For dephosphorylation assays, 10 units of Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units λ protein phosphatase (LPP) in a total reaction volume of 50 μl following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of SDS Page:Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units protein phosphatase (LPP) in a total reaction volume of 50 l following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of Western Blot:Article Title: PINK1 Kinase Catalytic Activity Is Regulated by Phosphorylation on Serines 228 and 402 Article Snippet: The mitochondrial lysate was incubated at 30 °C for 1 h with 200 units protein phosphatase (LPP) in a total reaction volume of 50 l following the instructions of the manufacturer (New England Biolabs). .. Purified PINK1 bound to FLAG beads was dephosphorylated either prior to or after the in vitro kinase assay by incubation at 30 °C for 15 min with 800 units of De-Phosphorylation Assay:Article Title: Phosphorylation of FANCD2 Inhibits the FANCD2/FANCI Complex and Suppresses the Fanconi Anemia Pathway in the Absence of DNA Damage Article Snippet: In vitro Phosphorylation and Dephosphorylation Assays Phosphorylation reaction containing FANCD2/FANCI and CK2 (P6010, NEB) were incubated at 30 C for 30 min in 50 mM Tris (pH 7.5), 10 mM MgCl2, 2 mM ATP and 2 mM DTT. .. Dephosphorylation reaction containing FANCD2/FANCI and Article Title: Ultrasonic Reporter of Kinase Activity Article Snippet: For all phosphorylation assays, 250 units of PKA (P6000L; NEB; specific activity: 1 pmol/min/unit), 1.5 μg of human CaMKIIα (Sino Biological; specific activity: 160 nmol/min/mg), or 0.1 μg of PKC alpha (Abcam; specific activity: 3,200 nmol/min/mg) per μL of engineered GVs at OD500 = 10 were mixed with 1 mM ATP (Sigma-Aldrich) in NEBufferTM for Protein Kinases (50 mM Tris-HCl, 10 mM MgCl 2 , 0.1 mM EDTA, 2 mM DTT, 0.01% Brij 35, pH 7.5; NEB) and incubated with slow rotation at 37°C for 10–12 hours, unless otherwise specified. .. For dephosphorylation assays, 10 units of Produced:Article Title: Untangling the interactions: Structural basis of nitric oxide regulated cyclic-di-GMP metabolism in bacteria Article Snippet: .. 70 pL of the supernatant was mixed with 10 uL of 3U/ml Mass Spectrometry:Article Title: Multisite phosphorylation dictates selective E2-E3 pairing as revealed by Ubc8/UBE2H-GID/CTLH assemblies. Article Snippet: .. Similarly, unlabeled bacterially expressed E2s were phosphorylated overnight at RT and analyzed with intact mass spectrometry (Figure 5D). |
