in an ethylenediaminetetraacetic acid tetrasodium salt dihydrate edta  (Millipore)


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    Name:
    Ethylenediaminetetraacetic acid tetrasodium salt dihydrate
    Description:
    Ethylenediaminetetraacetic acid EDTA is a chelator of metal ions It is a substituted diamine which has antibacterial activity EDTA removes the undesirable effects of ferric cupric and manganic ions in bleaching It prevents cellular division chlorophyll synthesis and algal biomass production EDTA is an inhibitor of metalloprotease It has anticoagulant property
    Catalog Number:
    e6511
    Price:
    None
    Applications:
    Ethylenediaminetetraacetic acid tetrasodium salt dihydrate has been used:. as a chelating agent to isolate human endometrial stem cell/stromal cells (hEnSCs) from menstrual blood. as a chelator in animal and testicular cell isolation. as a chelator for the culture of neural stem cells
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    Structured Review

    Millipore in an ethylenediaminetetraacetic acid tetrasodium salt dihydrate edta
    Ethylenediaminetetraacetic acid tetrasodium salt dihydrate
    Ethylenediaminetetraacetic acid EDTA is a chelator of metal ions It is a substituted diamine which has antibacterial activity EDTA removes the undesirable effects of ferric cupric and manganic ions in bleaching It prevents cellular division chlorophyll synthesis and algal biomass production EDTA is an inhibitor of metalloprotease It has anticoagulant property
    https://www.bioz.com/result/in an ethylenediaminetetraacetic acid tetrasodium salt dihydrate edta/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    in an ethylenediaminetetraacetic acid tetrasodium salt dihydrate edta - by Bioz Stars, 2020-05
    93/100 stars

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    Protease Inhibitor:

    Article Title: The polyamine transporter Slc18b1(VPAT) is important for both short and long time memory and for regulation of polyamine content in the brain
    Article Snippet: .. Acetone, ethylenediaminetetraacetic acid tetrasodium salt dihydrate (EDTA), protease inhibitor cocktail, phosphate buffered saline (PBS), Tris-HCl, diethylamide (DEA), trifluoroacetic acid (TFA), n-octyl-β-D-glucopyranoside, triethyl ammonium bicarbonate (TEAB), and formaldehyde CH2 O (37% (vol/vol)) were purchased from Sigma Aldrich (St. Louis, MO, USA). .. For tryptic digestion, iodoacetamide (IAA), urea and dithiothreitol (DTT) were obtained from Sigma Aldrich and trypsin (sequencing grade from bovine pancreas 1418475; Roche diagnostic, Basel, Switzerland) were used.

    Incubation:

    Article Title: Inhibition of In Vitro Fertilizing Capacity of Cryopreserved Mouse Sperm by Factors Released by Damaged Sperm, and Stimulation by Glutathione
    Article Snippet: .. The fertilized eggs were incubated overnight in drops of fertilization medium containing 0.1 mM EDTA (E6511, Sigma) under oil. ..

    other:

    Article Title: The yeast autophagy protease Atg4 is regulated by thioredoxin
    Article Snippet: The typical reaction mixture contained 4.5 μM Atg4, 5 μM Atg8, 1 mM 1,4-dithiothreitol (DTT; Sigma, 43815) and 1 mM EDTA (Sigma, E6511) in Tris-buffered saline (50 mM Trizma base, 138 mM NaCl, 27 mM KCl, pH 8).

    Article Title: Reduction of Articular and Systemic Inflammation by Kava-241 in a Porphyromonas gingivalis-Induced Arthritis Murine Model
    Article Snippet: Following fixation, specimens were consecutively washed with 5%, 10%, and 15% glycerol (American Bioanalytical, Natick, MA) in PBS, each for 15 min at 4°C, and decalcified in an EDTA tetrasodium salt dihydrate solution (Sigma-Aldrich, St. Louis, MO) for 14 days at 4°C.

    Article Title: Oxidation-stable amine-containing adsorbents for carbon dioxide capture
    Article Snippet: For the preparation of the aqueous solutions, trisodiumphosphate ( 1 in Fig. , Sigma Aldrich, 96%) and ethylenediaminetetraacetic acid tetrasodium salt dihydrate ( 4 , Sigma Aldrich, 99%) were directly dissolved in deionized water.

    Article Title: Detection of Numerous Y Chromosome Biallelic Polymorphisms by Denaturing High-Performance Liquid Chromatography
    Article Snippet: The mobile phase was 0.1 m triethylammonium acetate buffer at pH 7.0 (TEAA; cat. no. 400613, PE Applied Biosystems, Foster City, CA), containing in addition 0.1 m m tetrasodium ethylenediamine-tetraacetic acid (EDTA; cat. no. ED4SS, Sigma, St. Louis, MO).

    Molecular Weight:

    Article Title: Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp *
    Article Snippet: .. OTC hydrochloride and other TC antibiotics, bovine serum albumin (BSA), ovalbumin (OVA), complete and incomplete Freund’s adjuvants, polyethylene glycol (PEG) (molecular weight 3000–3700), 3,3',5,5'-tetramethylbenzidine (TMB), and ethylenediaminetetraacetic acid (EDTA) tetrasodium salt dihydrate were purchased from the Sigma Chemical Co. (MO, USA). .. Hybridoma serum-free medium was purchased from GIBCO (NY, USA).

    Cell Culture:

    Article Title: Inactivation of Retinoblastoma Protein (Rb1) in the Oocyte: Evidence That Dysregulated Follicle Growth Drives Ovarian Teratoma Formation in Mice
    Article Snippet: .. When no evidence of parthenogenesis was observed, we conducted a second series of experiments replicating a previously described culture protocol [ ] in which GV oocytes were collected and cultured in Minimal Essential Medium (MEM) with Earle’s balanced salt solution (Sigma, MO, USA) supplemented with essential amino acids (Sigma, MO, USA), Penicillin and Streptomycin (Gibco, Life Technologies, CA, USA), 0.01 mM Ethylenediaminetetraacetic acid tetrasodium salt hydrate (Tetrasodium EDTA, Sigma, USA), 0.23 mM pyruvic acid (Sigma, USA) and 3% bovine serum albumin (BSA, Sigma, USA). .. GV breakdown (GVBD), polar body (PB) extrusion and parthenogenetic activation were assessed as described previously [ ].

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    Millipore triton x 100 buffer
    Pretreating hippocampal slices with Trolox rescues Aβ 1–42 -induced GLT-1 Triton X-100 detergent-insolubility and ubiquitination.  A , Representative Western blots show GLT-1 levels in protein lysates first subjected to three serial 1% Triton X-100 extractions to remove Triton X-100-soluble proteins. Aβ 1–42  increased detergent-insoluble GLT-1 levels compared with control slices. Trolox reduced GLT-1 detergent-insolubility to control levels. Prestaining with Sypro confirmed equivalent protein loading among lanes.  B , Aβ 1–42  increased Triton X-100-insoluble GLT-1 (left) and was rescued by Trolox pretreatment (Ctrl, Aβ 1–42 , and Aβ 1–42  pretreated 1 h before and during 30 min Aβ 1–42  treatment;  n  = 9, 50, and 18, respectively;  F (2,74)  = 13.605,  p
    Triton X 100 Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 2 article reviews
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    85
    Millipore jak2 ip buffer
    Erythropoietin Receptor-Mediated Activation of the <t>JAK2/STAT5,</t> RAS/MAPK, and PI3K/AKT Signaling Pathways is Attenuated by BCR-ABL Kinase Activity in K562 Cells A. Normalized hEPO-mediated (10′ stimulation) JAK2 activation in K562 cells treated for 1hr with either 100nM dasatinib or 1μM imatinib. Data represents the average ± SD (n=3). JAK2 activation was monitored by immunoprecipitation and activation loop (Y1007) phosphorylation. JAK2 activation was normalized to the level of phospho-Y1007 observed in the “DMSO” condition for each experimental replicate. B. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of whole cell lysates from K562 cells after short-term and prolonged BCR-ABL inhibition (100nM dasatinib: 2hrs, 4hrs, 8hrs, 24hrs; 0.2% DMSO, 24hrs) followed by hEPO stimulation (10′). C. Normalized hEPO-mediated JAK2 activation in K562 cells after short-term (1hr) and prolonged (24hrs) 100nM dasatinib treatment followed hEPO stimulation (10′). Data is representative of triplicate experimental analysis. JAK2 activation and normalization was performed as in (A). D. Normalized RAS-GTP loading in K562 cells after short-term (1hr) and prolonged (24hrs) 100nM dasatinib treatment followed by hEPO stimulation (10′). Data is representative of triplicate experimental analysis. RAS activation was monitored using a RAS-GTP pulldown assay and RAS-GTP levels were normalized to the “DMSO” condition. E. Normalized hEPO-mediated (10′ stimulation) JAK2 activation in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 1μM imatinib, 500nM TG101348, dasatinib/TG101348, or imatinib/TG101348. JAK2 activation was monitored and normalized as in (A). F. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of whole cell lysates in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 1μM imatinib, 500nM TG101348, dasatinib/TG101348, or imatinib/TG101348 followed by hEPO stimulation (10′).
    Jak2 Ip Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 85 stars, based on 1 article reviews
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    99
    Millipore tris buffer
    Adenovirus-transduced artificial TDP-43 cytoplasmic aggregates have insoluble properties against various detergents. ( a, b ) <t>Sarkosyl-fractionated</t> cell lysates with <t>Tris</t> (T), 1% TritonX-100 (X), 1% sarkosyl (S) and 1 × SDS-sample (P) buffers were examined using phospho (p)-TDP-43 (pSer409/Ser410) ( a ) or C-terminal TDP-43 (405–414) antibody ( b ). ( c, d ) RIPA/urea-fractionated cell lysates with RIPA (R) and urea (U) buffers were examined using pTDP-43 ( c ) or C-terminal TDP-43 (405–414) antibody ( d ). The 74 and 52 kDa bands correspond to phosphorylated DsRed-tagged WT (←pWT) and CTF (←pCTF) TDP-43, respectively ( a, c ). The 72 and 50 kDa bands correspond to non-phosphorylated DsRed-tagged WT (←WT) and CTF (←CTF) TDP-43, respectively. Endogenous rat TDP-43 band at 43 kDa was indicated as arrowhead ( b, d ). ( a-d ) Full length gels and blots are shown.
    Tris Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pretreating hippocampal slices with Trolox rescues Aβ 1–42 -induced GLT-1 Triton X-100 detergent-insolubility and ubiquitination.  A , Representative Western blots show GLT-1 levels in protein lysates first subjected to three serial 1% Triton X-100 extractions to remove Triton X-100-soluble proteins. Aβ 1–42  increased detergent-insoluble GLT-1 levels compared with control slices. Trolox reduced GLT-1 detergent-insolubility to control levels. Prestaining with Sypro confirmed equivalent protein loading among lanes.  B , Aβ 1–42  increased Triton X-100-insoluble GLT-1 (left) and was rescued by Trolox pretreatment (Ctrl, Aβ 1–42 , and Aβ 1–42  pretreated 1 h before and during 30 min Aβ 1–42  treatment;  n  = 9, 50, and 18, respectively;  F (2,74)  = 13.605,  p

    Journal: The Journal of Neuroscience

    Article Title: Amyloid-?1–42 Slows Clearance of Synaptically Released Glutamate by Mislocalizing Astrocytic GLT-1

    doi: 10.1523/JNEUROSCI.5274-12.2013

    Figure Lengend Snippet: Pretreating hippocampal slices with Trolox rescues Aβ 1–42 -induced GLT-1 Triton X-100 detergent-insolubility and ubiquitination. A , Representative Western blots show GLT-1 levels in protein lysates first subjected to three serial 1% Triton X-100 extractions to remove Triton X-100-soluble proteins. Aβ 1–42 increased detergent-insoluble GLT-1 levels compared with control slices. Trolox reduced GLT-1 detergent-insolubility to control levels. Prestaining with Sypro confirmed equivalent protein loading among lanes. B , Aβ 1–42 increased Triton X-100-insoluble GLT-1 (left) and was rescued by Trolox pretreatment (Ctrl, Aβ 1–42 , and Aβ 1–42 pretreated 1 h before and during 30 min Aβ 1–42 treatment; n = 9, 50, and 18, respectively; F (2,74) = 13.605, p

    Article Snippet: Protein lysates from slices and cultured astrocytes were prepared using 1% Triton X-100 buffer [in m m : 20 Tris, 1 EDTA, 0.5 EGTA, 250 sucrose, Protease Inhibitor Cocktail (EMD Millipore)].

    Techniques: Western Blot

    Erythropoietin Receptor-Mediated Activation of the JAK2/STAT5, RAS/MAPK, and PI3K/AKT Signaling Pathways is Attenuated by BCR-ABL Kinase Activity in K562 Cells A. Normalized hEPO-mediated (10′ stimulation) JAK2 activation in K562 cells treated for 1hr with either 100nM dasatinib or 1μM imatinib. Data represents the average ± SD (n=3). JAK2 activation was monitored by immunoprecipitation and activation loop (Y1007) phosphorylation. JAK2 activation was normalized to the level of phospho-Y1007 observed in the “DMSO” condition for each experimental replicate. B. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of whole cell lysates from K562 cells after short-term and prolonged BCR-ABL inhibition (100nM dasatinib: 2hrs, 4hrs, 8hrs, 24hrs; 0.2% DMSO, 24hrs) followed by hEPO stimulation (10′). C. Normalized hEPO-mediated JAK2 activation in K562 cells after short-term (1hr) and prolonged (24hrs) 100nM dasatinib treatment followed hEPO stimulation (10′). Data is representative of triplicate experimental analysis. JAK2 activation and normalization was performed as in (A). D. Normalized RAS-GTP loading in K562 cells after short-term (1hr) and prolonged (24hrs) 100nM dasatinib treatment followed by hEPO stimulation (10′). Data is representative of triplicate experimental analysis. RAS activation was monitored using a RAS-GTP pulldown assay and RAS-GTP levels were normalized to the “DMSO” condition. E. Normalized hEPO-mediated (10′ stimulation) JAK2 activation in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 1μM imatinib, 500nM TG101348, dasatinib/TG101348, or imatinib/TG101348. JAK2 activation was monitored and normalized as in (A). F. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of whole cell lysates in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 1μM imatinib, 500nM TG101348, dasatinib/TG101348, or imatinib/TG101348 followed by hEPO stimulation (10′).

    Journal: Cancer discovery

    Article Title: MEK-Dependent Negative Feedback Underlies BCR-ABL-Mediated Oncogene Addiction

    doi: 10.1158/2159-8290.CD-13-0235

    Figure Lengend Snippet: Erythropoietin Receptor-Mediated Activation of the JAK2/STAT5, RAS/MAPK, and PI3K/AKT Signaling Pathways is Attenuated by BCR-ABL Kinase Activity in K562 Cells A. Normalized hEPO-mediated (10′ stimulation) JAK2 activation in K562 cells treated for 1hr with either 100nM dasatinib or 1μM imatinib. Data represents the average ± SD (n=3). JAK2 activation was monitored by immunoprecipitation and activation loop (Y1007) phosphorylation. JAK2 activation was normalized to the level of phospho-Y1007 observed in the “DMSO” condition for each experimental replicate. B. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of whole cell lysates from K562 cells after short-term and prolonged BCR-ABL inhibition (100nM dasatinib: 2hrs, 4hrs, 8hrs, 24hrs; 0.2% DMSO, 24hrs) followed by hEPO stimulation (10′). C. Normalized hEPO-mediated JAK2 activation in K562 cells after short-term (1hr) and prolonged (24hrs) 100nM dasatinib treatment followed hEPO stimulation (10′). Data is representative of triplicate experimental analysis. JAK2 activation and normalization was performed as in (A). D. Normalized RAS-GTP loading in K562 cells after short-term (1hr) and prolonged (24hrs) 100nM dasatinib treatment followed by hEPO stimulation (10′). Data is representative of triplicate experimental analysis. RAS activation was monitored using a RAS-GTP pulldown assay and RAS-GTP levels were normalized to the “DMSO” condition. E. Normalized hEPO-mediated (10′ stimulation) JAK2 activation in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 1μM imatinib, 500nM TG101348, dasatinib/TG101348, or imatinib/TG101348. JAK2 activation was monitored and normalized as in (A). F. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of whole cell lysates in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 1μM imatinib, 500nM TG101348, dasatinib/TG101348, or imatinib/TG101348 followed by hEPO stimulation (10′).

    Article Snippet: Cells were lysed in JAK2 IP buffer (50mM Tris pH 7.6, 100mM NaCl, 1mM EDTA, 1mM EGTA, 0.5% NP40, 0.1% Triton) or RAS IP buffer (50mM Tris pH 7.5, 125mM NaCl, 6.5mM MgCl2 , 5% glycerol, 0.2% NP40) supplemented with 1% protease and 1% phosphatase inhibitors (Calbiochem).

    Techniques: Activation Assay, Activity Assay, Immunoprecipitation, Western Blot, Inhibition

    Global Gene Expression Analysis of Dasatinib Treated K562 Cells Identifies Candidate Mediators Responsible for the Attenuation of Myeloid GF-R Signaling in CML Cells A. ). The heat map at the bottom highlights a few of the genes with increased expression following dasatinib treatment in K562 cells. B. Quantitative PCR (qPCR) analysis of potential negative feedback genes in K562 cells treated with dasatinib (100nM), imatinib (1μM), or PD0325901 (500nM). The average fold expression change (2 (−ΔΔCt) ) and standard deviation (SD fold-change ) for each gene is represented (n=3). C. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of RAS and ERK activity before and after hEPO-stimulation (10′) in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 500nM PD0325901, or dasatinib/PD0325901. RAS activity was monitored using a RAS-GTP pulldown assay. D. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of JAK2 and STAT5 activity before and after hEPO-stimulation in K562 cells treated under the same experimental conditions as (A). JAK2 activation was determined by immunoprecipitation and activation loop (Y1007) phosphorylation.

    Journal: Cancer discovery

    Article Title: MEK-Dependent Negative Feedback Underlies BCR-ABL-Mediated Oncogene Addiction

    doi: 10.1158/2159-8290.CD-13-0235

    Figure Lengend Snippet: Global Gene Expression Analysis of Dasatinib Treated K562 Cells Identifies Candidate Mediators Responsible for the Attenuation of Myeloid GF-R Signaling in CML Cells A. ). The heat map at the bottom highlights a few of the genes with increased expression following dasatinib treatment in K562 cells. B. Quantitative PCR (qPCR) analysis of potential negative feedback genes in K562 cells treated with dasatinib (100nM), imatinib (1μM), or PD0325901 (500nM). The average fold expression change (2 (−ΔΔCt) ) and standard deviation (SD fold-change ) for each gene is represented (n=3). C. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of RAS and ERK activity before and after hEPO-stimulation (10′) in K562 cells pretreated for 24hrs with 0.2% DMSO, 100nM dasatinib, 500nM PD0325901, or dasatinib/PD0325901. RAS activity was monitored using a RAS-GTP pulldown assay. D. Upper: Line diagram representation of duration of serum starve, kinase inhibitor treatment, and growth-factor stimulation. Lower: Western immunoblot analysis of JAK2 and STAT5 activity before and after hEPO-stimulation in K562 cells treated under the same experimental conditions as (A). JAK2 activation was determined by immunoprecipitation and activation loop (Y1007) phosphorylation.

    Article Snippet: Cells were lysed in JAK2 IP buffer (50mM Tris pH 7.6, 100mM NaCl, 1mM EDTA, 1mM EGTA, 0.5% NP40, 0.1% Triton) or RAS IP buffer (50mM Tris pH 7.5, 125mM NaCl, 6.5mM MgCl2 , 5% glycerol, 0.2% NP40) supplemented with 1% protease and 1% phosphatase inhibitors (Calbiochem).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation, Western Blot, Activity Assay, Activation Assay, Immunoprecipitation

    Adenovirus-transduced artificial TDP-43 cytoplasmic aggregates have insoluble properties against various detergents. ( a, b ) Sarkosyl-fractionated cell lysates with Tris (T), 1% TritonX-100 (X), 1% sarkosyl (S) and 1 × SDS-sample (P) buffers were examined using phospho (p)-TDP-43 (pSer409/Ser410) ( a ) or C-terminal TDP-43 (405–414) antibody ( b ). ( c, d ) RIPA/urea-fractionated cell lysates with RIPA (R) and urea (U) buffers were examined using pTDP-43 ( c ) or C-terminal TDP-43 (405–414) antibody ( d ). The 74 and 52 kDa bands correspond to phosphorylated DsRed-tagged WT (←pWT) and CTF (←pCTF) TDP-43, respectively ( a, c ). The 72 and 50 kDa bands correspond to non-phosphorylated DsRed-tagged WT (←WT) and CTF (←CTF) TDP-43, respectively. Endogenous rat TDP-43 band at 43 kDa was indicated as arrowhead ( b, d ). ( a-d ) Full length gels and blots are shown.

    Journal: PLoS ONE

    Article Title: Formation and spreading of TDP-43 aggregates in cultured neuronal and glial cells demonstrated by time-lapse imaging

    doi: 10.1371/journal.pone.0179375

    Figure Lengend Snippet: Adenovirus-transduced artificial TDP-43 cytoplasmic aggregates have insoluble properties against various detergents. ( a, b ) Sarkosyl-fractionated cell lysates with Tris (T), 1% TritonX-100 (X), 1% sarkosyl (S) and 1 × SDS-sample (P) buffers were examined using phospho (p)-TDP-43 (pSer409/Ser410) ( a ) or C-terminal TDP-43 (405–414) antibody ( b ). ( c, d ) RIPA/urea-fractionated cell lysates with RIPA (R) and urea (U) buffers were examined using pTDP-43 ( c ) or C-terminal TDP-43 (405–414) antibody ( d ). The 74 and 52 kDa bands correspond to phosphorylated DsRed-tagged WT (←pWT) and CTF (←pCTF) TDP-43, respectively ( a, c ). The 72 and 50 kDa bands correspond to non-phosphorylated DsRed-tagged WT (←WT) and CTF (←CTF) TDP-43, respectively. Endogenous rat TDP-43 band at 43 kDa was indicated as arrowhead ( b, d ). ( a-d ) Full length gels and blots are shown.

    Article Snippet: Cell fractionation For sarkosyl fractionation, cells were lysed and sonicated in Tris buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM EDTA and 5 mM EGTA) containing phosphatase inhibitors (5 mM NaF and 2 mM Na3 VO4 ) and 1× protease inhibitor cocktail (Millipore, Billerica, MA, USA), followed by centrifugation at 160,000 × g for 20 min at 4°C.

    Techniques: