Activation of endogenous protein phosphatase 1 enhances the calcium sensitivity of the ryanodine receptor type 2 in murine ventricular cardiomyocytes.

KEY POINTS
Increased protein phosphatase 1 (PP-1) activity was found in the late stages of human heart failure. Although PP-1 has been studied extensively, a detailed understanding of its role in excitation-contraction coupling mechanism, in normal and diseased heart, remains elusive. This work examines the functional effects of the activities of PP-1 on the release of Ca2 + local events in ventricular cardiomyocytes, using the activation peptide (PDP3) for the stimulation of endogenous PP-1 protein. We reported that the acute de-phosphorylation may increase the sensitivity of RyR2 to Ca2 + channels in situ, and that RyR2-serine2808 phosphorylation sites can mediate the process. Our approach unmask the functional importance of PP-1 in the regulation of RyR2 activity, suggesting a potential role in the generation of the pathophysiology of the sarcoplasmic reticulum Ca2 + leak at the heart sick.


ABSTRACT
Changes heart Ryanodine receptor (RyR2) phosphorylation was considered a disease of regulation and post-translational modification of proteins related matters. The extent of phosphorylation of RyR2 is mainly determined by the balance of the activity of protein kinases and phosphatases, respectively. Increased protein phosphatase-1 (PP-1) activity has been observed in heart failure, but the role of this enzyme in the regulation of intracellular Ca2 + handling the remains poorly understood.

To determine the physiological and significance of the pathophysiology of increased activity of PP-1, we investigate how the PP-1 catalytic subunit (PP-1c) alter Ca2 + sparks in cardiomyocytes permeabilized and we also apply PP-1-disturbing peptide (PDP3) to specifically activate endogenous PP-1, including one anchored in the RyR2 macromolecular complex.

We compared the wild type (WT) and transgenic mice in which the normally highly phosphorylated RyR2 site-S2808 has been ablated to investigate his involvement in RyR2 modulation (S2808A + / +). In WT myocytes, PP-1 increased the frequency of Ca2 + spark (CaSpF) by 2-fold, followed by depletion of sarcoplasmic reticulum (SR) Ca2 + stores. Similarly, PDP3 temporarily increase and decrease the frequency spark SR Ca2 + load. RyR2 Ca2 + sensitivity, as assessed by analysis of Ca2 + spark recovery, increase with PDP3 when compared to the negative control peptide. S2808A + / + cardiomyocytes do not respond well PP-1c and treatment PDP3.

Our results showed an increase in Ca 2+ sensitivity of RyR2 in the de-phosphorylation by PP-1. Furthermore, we confirmed the S2808 as a target site for PP-1 and as a potential relationship between the modulation RyR2s and cellular responses. This article is protected by copyright. All rights reserved.

Activation of endogenous <em>protein</em> <em>phosphatase</em> <em>1</em> enhances the calcium sensitivity of the ryanodine receptor type 2 in murine ventricular cardiomyocytes.
Activation of endogenous protein phosphatase 1 enhances the calcium sensitivity of the ryanodine receptor type 2 in murine ventricular cardiomyocytes.

Phactr one set Slack (KCNT one ) channels through protein phosphatase one (PP one ).

The Slack (KCNT1) gene encodes sodium-activated potassium channels are abundant expressed in the central nervous system. human mutations alter channel function Slack, resulting in epilepsy and intellectual disability. The majority of disease-causing mutations located in the cytoplasmic C-terminus extended channels and the result is increased current Slack Slack. Previous experiments have shown that the C-terminus of the protein channel Slack bind a number of cytoplasmic signaling.

PPP3R1 siRNA

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Description: Rabbit polyclonal PPP3R1 antibody

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PPP3R1 Antibody

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Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human. This antibody is Unconjugated. Tested in the following application: ELISA, WB; Recommended dilution: WB:1:500-1:2000

PPP3R1 Antibody

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Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.

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Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.

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EUR 349.3
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.

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V8030-20UG 20 ug
EUR 153.3
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.

Calcineurin B Antibody / PPP3R1

V8030SAF-100UG 100 ug
EUR 349.3
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.

PPP3R1 Antibody, FITC conjugated

1-CSB-PA018575LC01HU
  • EUR 380.40
  • EUR 402.00
  • 100ug
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Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human. This antibody is FITC conjugated. Tested in the following application: ELISA

OACD06670-1ML - PPP3R1 Antibody

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PPP3R1 Recombinant Protein (Mouse)

RP164066 100 ug Ask for price

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PPP3R1 Antibody, Biotin conjugated

1-CSB-PA018575LD01HU
  • EUR 380.40
  • EUR 402.00
  • 100ug
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Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA

OACD06670-100UL - PPP3R1 Antibody

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PPP3R1 (GFP-tagged) - Human protein phosphatase 3, regulatory subunit B, alpha (PPP3R1)

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Ppp3r1 ORF Vector (Rat) (pORF)

ORF073949 1.0 ug DNA
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[One Step] PPP3R1 Antibody Kit

RK05683 50 ul
EUR 288

Human PPP3R1 Protein, His Tag

E40KMPH4377 20ug
EUR 495

PPP3R1 ELISA Kit (Rat) (OKEH00948)

OKEH00948 96 Wells
EUR 794.4
Description: Description of target: regulatory subunit of the enzyme that catalyzes the calcium-dependent removal of serine- and threonine-bound phosphate groups [RGD, Feb 2006];Species reactivity: Rat;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.086 ng/mL

PPP3R1 (Myc-DDK-tagged)-Human protein phosphatase 3, regulatory subunit B, alpha (PPP3R1)

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Human PPP3R1 knockout cell line

ABC-KH12044 1 vial Ask for price
Description: Human PPP3R1 knockout cell line is HEK293/HeLa cell line, edited by CRISPR/Cas9 technology.

PPP3R1 ELISA Kit (Human) (OKEH00947)

OKEH00947 96 Wells
EUR 794.4
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PPP3R1 ELISA Kit (Mouse) (OKEH07121)

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EUR 794.4
Description: Description of target: Regulatory subunit of calcineurin, a calcium-dependent, calmodulin stimulated protein phosphatase. Confers calcium sensitivity. ;Species reactivity: Mouse;Application: ;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.085 ng/mL

PPP3R1 ELISA Kit (Mouse) (OKCD02818)

OKCD02818 96 Wells
EUR 1028.4
Description: Description of target: Regulatory subunit of calcineurin, a calcium-dependent, calmodulin stimulated protein phosphatase. Confers calcium sensitivity. ;Species reactivity: Mouse;Application: ;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.047 ng/mL

Human PPP3R1 knockdown cell line

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EUR 213
Description: Human PPP3R1 Protein Lysate 20ug

OCOA17193-20UG - PPP3R1 Protein Lysate

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PPP3R1 Polyclonal Antibody, HRP Conjugated

A60397
  • EUR 684.66
  • EUR 302.50
  • EUR 423.50
  • 100 µg
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PPP3R1 Polyclonal Antibody, FITC Conjugated

A60396
  • EUR 684.66
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  • EUR 423.50
  • 100 µg
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OPCD06358-10UG - PPP3R1 Recombinant Protein

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EUR 129

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OPCD06358-50UG 50ug
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Calcineurin B / PPP3R1(BE2.1), 0.2mg/mL

BNUB2321-100 1uL
EUR 225
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Calcineurin B / PPP3R1(BE2.1), 0.2mg/mL

BNUB2321-500 1uL
EUR 485
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), 1mg/mL

BNUM2321-50 1uL
EUR 396
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

PPP3R1 Polyclonal Antibody, Biotin Conjugated

A60395
  • EUR 684.66
  • EUR 302.50
  • EUR 423.50
  • 100 µg
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OPCD06358-200UG - PPP3R1 Recombinant Protein

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EUR 639

OKEH00948-96W - PPP3R1 ELISA Kit (Rat)

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EUR 525

OKEH07121-96W - PPP3R1 ELISA Kit (Mouse)

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OKCD02818-96W - PPP3R1 ELISA Kit (Mouse)

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EUR 700

OKEH00947-96W - PPP3R1 ELISA Kit (Human)

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EUR 525

Ppp3r1 sgRNA CRISPR Lentivector set (Rat)

K6874001 3 x 1.0 ug
EUR 406.8

PPP3R1 sgRNA CRISPR Lentivector set (Human)

K1709301 3 x 1.0 ug
EUR 406.8

Ppp3r1 3'UTR GFP Stable Cell Line

TU266693 1.0 ml Ask for price

PPP3R1 3'UTR GFP Stable Cell Line

TU068697 1.0 ml
EUR 1825.2

Calcineurin B / PPP3R1(CALNB/2342), 0.2mg/mL

BNUB2342-100 1uL
EUR 225
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(CALNB/2342), 0.2mg/mL

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EUR 485
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Calcineurin B / PPP3R1(CALNB/2342), 1mg/mL

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EUR 396
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

PPP3R1 Protein Vector (Rat) (pPM-C-HA)

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PPP3R1 Protein Vector (Rat) (pPB-C-His)

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Ppp3r1 3'UTR Luciferase Stable Cell Line

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PPP3R1 3'UTR Luciferase Stable Cell Line

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Ppp3r1 (untagged) - Mouse protein phosphatase 3, regulatory subunit B, alpha isoform (calcineurin B, type I) (Ppp3r1), (10ug)

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Recombinant Human CNB/PPP3R1 Protein, His Tag

E40KMH895 20ug
EUR 495

Ppp3r1 sgRNA CRISPR Lentivector (Rat) (Target 1)

K6874002 1.0 ug DNA
EUR 184.8

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EUR 99

Calcineurin Subunit B Type 1 (PPP3R1) Antibody

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EUR 577.2

Calcineurin Subunit B Type 1 (PPP3R1) Antibody

20-abx217914
  • EUR 510.00
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  • 100 ug
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Calcineurin Subunit B Type 1 (PPP3R1) Antibody

20-abx114854
  • EUR 878.40
  • EUR 477.60
  • 150 ul
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Calcineurin Subunit B Type 1 (PPP3R1) Antibody

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EUR 469.2

Calcineurin Subunit B Type 1 (PPP3R1) Antibody

20-abx000913
  • EUR 493.20
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  • 100 ul
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Calcineurin Subunit B Type 1 (PPP3R1) Antibody

20-abx302644
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
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PPP3R1 sgRNA CRISPR Lentivector (Human) (Target 1)

K1709302 1.0 ug DNA
EUR 184.8

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EUR 3308.4

Recombinant Human PPP3R1 Protein, His, E.coli-5ug

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EUR 186

Human PPP3R1 Over-expressing Stable Cell Line

ABC-X2418 1 vial Ask for price
Description: Gentaur can provide custom lentiviral constructs expressing any genes of interest as long as it is less than about 3 kb. Lentiviral technology enables us to efficiently generate stable expression lines which are then selected for moderate or high expressers, depending on the experimental requirements. If you are interested in specific lentiviral DNA constructs or have further questions, please contact us to discuss the details. PPP3R1 protein phosphatase 3, regulatory subunit B, alpha [ Homo sapiens ] http://www.ncbi.nlm.nih.gov/gene/5534

Recombinant Human PPP3R1 Protein, His, E.coli-20ug

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EUR 241.2

Ppp3r1 (Myc-DDK-tagged) - Mouse protein phosphatase 3, regulatory subunit B, alpha isoform (calcineurin B, type I) (Ppp3r1)

MR227456 10 µg Ask for price

Ppp3r1 (GFP-tagged) - Mouse protein phosphatase 3 regulatory subunit B alpha isoform (calcineurin B type I) (Ppp3r1), (10ug)

MG227456 10 µg Ask for price

Monoclonal PPP3R1 Antibody (monoclonal) (M01), Clone: 40

AMM07304G 0.1mg
EUR 580.8
Description: A Monoclonal antibody against Human PPP3R1 (monoclonal) (M01). The antibodies are raised in mouse and are from clone 40. This antibody is applicable in WB and IF, E

Polyclonal PPP3R1 / Calcineurin B Antibody (aa7-37)

AMM07318G 0.05mg
EUR 580.8
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human PPP3R1 / Calcineurin B (aa7-37). This antibody is tested and proven to work in the following applications:

Lenti ORF particles, PPP3R1 (mGFP-tagged) - Human protein phosphatase 3, regulatory subunit B, alpha (PPP3R1), 200ul, >10^7 TU/mL

RC206585L4V 200 µl Ask for price

AAP56124-100UG - PPP3R1 Peptide - N-terminal region

AAP56124-100UG 100ug
EUR 99

PPP3R1 Recombinant Protein N-6His Tag Lyophilized

IHUPPP3R1RN6HISLY10UG each
EUR 203
Description: PPP3R1 Recombinant Protein N-6His Tag Lyophilized

Ppp3r1 (untagged ORF) - Rat protein phosphatase 3, regulatory subunit B, alpha isoform (calcineurin B, type I) (Ppp3r1), (10 ug)

RN207229 10 µg Ask for price

Lenti ORF particles, PPP3R1 (Myc-DDK tagged) - Human protein phosphatase 3, regulatory subunit B, alpha (PPP3R1), 200ul, >10^7 TU/mL

RC206585L3V 200 µl Ask for price

ARP56124_P050 - PPP3R1 antibody - N-terminal region (ARP56124_P050)

ARP56124_P050 100ul
EUR 389

Calcineurin B / PPP3R1(BE2.1), CF568 conjugate, 0.1mg/mL

BNC682321-100 1uL
EUR 192
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF568 conjugate, 0.1mg/mL

BNC682321-500 1uL
EUR 532
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF647 conjugate, 0.1mg/mL

BNC472321-100 1uL
EUR 192
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF647 conjugate, 0.1mg/mL

BNC472321-500 1uL
EUR 532
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF594 conjugate, 0.1mg/mL

BNC942321-100 1uL
EUR 192
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF594 conjugate, 0.1mg/mL

BNC942321-500 1uL
EUR 532
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF640R conjugate, 0.1mg/mL

BNC402321-100 1uL
EUR 192
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF640R conjugate, 0.1mg/mL

BNC402321-500 1uL
EUR 532
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF405S conjugate, 0.1mg/mL

BNC042321-100 1uL
EUR 192
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF405S conjugate, 0.1mg/mL

BNC042321-500 1uL
EUR 532
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF488A conjugate, 0.1mg/mL

BNC882321-100 1uL
EUR 192
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), CF488A conjugate, 0.1mg/mL

BNC882321-500 1uL
EUR 532
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

Calcineurin B / PPP3R1(BE2.1), Biotin conjugate, 0.1mg/mL

BNCB2321-100 1uL
EUR 192
Description: Primary and secondary antibodies for multiple methodologyimmunostaining detection application

One is Phactr1, actin-binding protein that recruits protein phosphatase 1 (PP1) to specific phosphoprotein substrates. Using co-immunoprecipitation, we found that Phactr1 required to connect the channel to actin. Using a clamp recordings, we found that co-expression of wild-type channels Phactr1 with Slack reduce current amplitude but had no effect on the channel Slack where preserved PKC phosphorylation sites (S407), which regulates the current amplitude has been mutated.