sodium pervanadate  (Millipore)


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    Structured Review

    Millipore sodium pervanadate
    Fig. 7. ( A ) Kv2.1 preferentially associates with the D245A cyt-PTPε substrate-trapping mutant. HEK 293 cells were transiently transfected with wild-type cyt-PTPε, D245A cyt-PTPε, Kv2.1, or combinations thereof. Following lysis, PTPε was immunoprecipitated via its attached FLAG tag; associated Kv2.1 was detected by protein blot analysis of immunoprecipitates (top panel). The second, third and fourth panels document expression of cyt-PTPε and Kv2.1 and immunoprecipitation of cyt-PTPε in the various transfections. The presence of <t>pervanadate</t> (right-most lane) somewhat reduces the electrophoretic mobility of some of the proteins examined, most likely by more effectively reducing their dephosphorylation by phosphatases present in cell lysates during precipitation. ( B ) Expression and activities of Src and Fyn are unaltered in Ptpre –/– Schwann cells. Total protein from primary Schwann cells of 3- to 5-day-old mice were immunoprecipitated with anti-Src or anti-Fyn antibodies. A fraction of the precipitated kinases was allowed to phosphorylate acid-denatured enolase in vitro in the presence of [γ- 32 P]ATP, followed by SDS–PAGE analysis and exposure to film (top panels). Another fraction of the immunoprecipitated material was analyzed by protein blotting for expression levels of Src or Fyn (bottom panels). Tyrosine phosphorylation levels of both kinases were unaltered (not shown). The experiment shown is representative of three performed.
    Sodium Pervanadate, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sodium pervanadate/product/Millipore
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    sodium pervanadate - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Hypomyelination and increased activity of voltage-gated K+ channels in mice lacking protein tyrosine phosphatase ?"

    Article Title: Hypomyelination and increased activity of voltage-gated K+ channels in mice lacking protein tyrosine phosphatase ?

    Journal: The EMBO Journal

    doi: 10.1093/emboj/19.15.4036

    Fig. 7. ( A ) Kv2.1 preferentially associates with the D245A cyt-PTPε substrate-trapping mutant. HEK 293 cells were transiently transfected with wild-type cyt-PTPε, D245A cyt-PTPε, Kv2.1, or combinations thereof. Following lysis, PTPε was immunoprecipitated via its attached FLAG tag; associated Kv2.1 was detected by protein blot analysis of immunoprecipitates (top panel). The second, third and fourth panels document expression of cyt-PTPε and Kv2.1 and immunoprecipitation of cyt-PTPε in the various transfections. The presence of pervanadate (right-most lane) somewhat reduces the electrophoretic mobility of some of the proteins examined, most likely by more effectively reducing their dephosphorylation by phosphatases present in cell lysates during precipitation. ( B ) Expression and activities of Src and Fyn are unaltered in Ptpre –/– Schwann cells. Total protein from primary Schwann cells of 3- to 5-day-old mice were immunoprecipitated with anti-Src or anti-Fyn antibodies. A fraction of the precipitated kinases was allowed to phosphorylate acid-denatured enolase in vitro in the presence of [γ- 32 P]ATP, followed by SDS–PAGE analysis and exposure to film (top panels). Another fraction of the immunoprecipitated material was analyzed by protein blotting for expression levels of Src or Fyn (bottom panels). Tyrosine phosphorylation levels of both kinases were unaltered (not shown). The experiment shown is representative of three performed.
    Figure Legend Snippet: Fig. 7. ( A ) Kv2.1 preferentially associates with the D245A cyt-PTPε substrate-trapping mutant. HEK 293 cells were transiently transfected with wild-type cyt-PTPε, D245A cyt-PTPε, Kv2.1, or combinations thereof. Following lysis, PTPε was immunoprecipitated via its attached FLAG tag; associated Kv2.1 was detected by protein blot analysis of immunoprecipitates (top panel). The second, third and fourth panels document expression of cyt-PTPε and Kv2.1 and immunoprecipitation of cyt-PTPε in the various transfections. The presence of pervanadate (right-most lane) somewhat reduces the electrophoretic mobility of some of the proteins examined, most likely by more effectively reducing their dephosphorylation by phosphatases present in cell lysates during precipitation. ( B ) Expression and activities of Src and Fyn are unaltered in Ptpre –/– Schwann cells. Total protein from primary Schwann cells of 3- to 5-day-old mice were immunoprecipitated with anti-Src or anti-Fyn antibodies. A fraction of the precipitated kinases was allowed to phosphorylate acid-denatured enolase in vitro in the presence of [γ- 32 P]ATP, followed by SDS–PAGE analysis and exposure to film (top panels). Another fraction of the immunoprecipitated material was analyzed by protein blotting for expression levels of Src or Fyn (bottom panels). Tyrosine phosphorylation levels of both kinases were unaltered (not shown). The experiment shown is representative of three performed.

    Techniques Used: Mutagenesis, Transfection, Lysis, Immunoprecipitation, FLAG-tag, Expressing, De-Phosphorylation Assay, Mouse Assay, In Vitro, SDS Page

    2) Product Images from "Epitope-specific crosslinking of CD45 down-regulates membrane-associated tyrosine phosphatase activity and triggers early signalling events in human activated T cells"

    Article Title: Epitope-specific crosslinking of CD45 down-regulates membrane-associated tyrosine phosphatase activity and triggers early signalling events in human activated T cells

    Journal: Immunology

    doi: 10.1111/j.1365-2567.2004.01986.x

    Broad spectrum PTPase inhibitors block TNF-α gene expression. Primary activated T cells (3 × 10 6 ) were preincubated for 30 min (a) with PTPase inhibitors PAO, or (b) with sodium pervanadate (Na 3 VO 4 /H 2 O 2 ), then stimulated for 2 hr with anti-CD45 mAb 4B2 (20 µg/ml)crosslinked on plastic. (c) Primary activated T cells were incubated (30 min) with the serine/threonine phosphatase inhibitor okadaic acid prior to stimulation with anti-CD45 mAb 4B2 as for (a) and (b), or left unstimulated (unstim.). Total RNA was isolated and analysed by RT-PCR using TNF-α-specific primers. Results are expressed as fold induction over unstimulated T cells. Similar results were obtained with cells stimulated with mAb 9.4.
    Figure Legend Snippet: Broad spectrum PTPase inhibitors block TNF-α gene expression. Primary activated T cells (3 × 10 6 ) were preincubated for 30 min (a) with PTPase inhibitors PAO, or (b) with sodium pervanadate (Na 3 VO 4 /H 2 O 2 ), then stimulated for 2 hr with anti-CD45 mAb 4B2 (20 µg/ml)crosslinked on plastic. (c) Primary activated T cells were incubated (30 min) with the serine/threonine phosphatase inhibitor okadaic acid prior to stimulation with anti-CD45 mAb 4B2 as for (a) and (b), or left unstimulated (unstim.). Total RNA was isolated and analysed by RT-PCR using TNF-α-specific primers. Results are expressed as fold induction over unstimulated T cells. Similar results were obtained with cells stimulated with mAb 9.4.

    Techniques Used: Blocking Assay, Expressing, Incubation, Isolation, Reverse Transcription Polymerase Chain Reaction

    3) Product Images from "Molecular Networks in FGF Signaling: Flotillin-1 and Cbl-Associated Protein Compete for the Binding to Fibroblast Growth Factor Receptor Substrate 2"

    Article Title: Molecular Networks in FGF Signaling: Flotillin-1 and Cbl-Associated Protein Compete for the Binding to Fibroblast Growth Factor Receptor Substrate 2

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0029739

    Flotillin-1 is required for the recruitment of FRS2 into light membranes in pervanadate treated cells. Hep3B cells (control: upper panels, flot-1 knockdown: lower panels) were starved overnight and then stimulated with pervanadate. Detergent resistant light membranes were prepared using density gradient centrifugation and found in fractions 1–3 of the gradient. The localization of FRS2, flot-1 and CAP was analyzed. Western blots for transferrin receptor (TfnR), GAPDH and GM1-bound cholera toxin subunit B (CTX-B) were used to control the gradient.
    Figure Legend Snippet: Flotillin-1 is required for the recruitment of FRS2 into light membranes in pervanadate treated cells. Hep3B cells (control: upper panels, flot-1 knockdown: lower panels) were starved overnight and then stimulated with pervanadate. Detergent resistant light membranes were prepared using density gradient centrifugation and found in fractions 1–3 of the gradient. The localization of FRS2, flot-1 and CAP was analyzed. Western blots for transferrin receptor (TfnR), GAPDH and GM1-bound cholera toxin subunit B (CTX-B) were used to control the gradient.

    Techniques Used: Gradient Centrifugation, Western Blot

    4) Product Images from "PDGFRβ translocates to the nucleus and regulates chromatin remodeling via TATA element–modifying factor 1"

    Article Title: PDGFRβ translocates to the nucleus and regulates chromatin remodeling via TATA element–modifying factor 1

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201706118

    PDGF-BB–induced nuclear transport of PDGFRβ is not quenched by receptor kinase inhibition. (a) PDGFRβ translocation to the nucleus upon inhibition of PDGFRβ kinase activity. Nuclear extracts were immunoblotted for pTyr857-PDGFRβ (top) or immunoprecipitated and immunoblotted with the PDGFRβ antibody ctβ (second panel). Cytoplasmic extracts were immunoblotted for PDGFRβ with the ctβ antibody (third panel) or α-tubulin (bottom). (b) Quantification of PDGFRβ distribution upon inhibition of receptor tyrosine kinase activity with AG1296. Immunoblots were quantified as described in Fig. 1 . Error bars indicate SD. Quantification was based on four independent experiments. (c) Cells were stimulated with PDGF-BB or phosphatase activity was inhibited with sodium pervanadate, and lysates were immunoblotted with total PDGFR antibody (Y92) or lamin A/C for nuclear fractions (top two panels) or total PDGFRβ, α-tubulin, and pPDGFRβ antibody for cytoplasmic fractions (three bottom panels). Molecular mass was measured in kilodaltons. IB, immunoblotting; IP, immunoprecipitation.
    Figure Legend Snippet: PDGF-BB–induced nuclear transport of PDGFRβ is not quenched by receptor kinase inhibition. (a) PDGFRβ translocation to the nucleus upon inhibition of PDGFRβ kinase activity. Nuclear extracts were immunoblotted for pTyr857-PDGFRβ (top) or immunoprecipitated and immunoblotted with the PDGFRβ antibody ctβ (second panel). Cytoplasmic extracts were immunoblotted for PDGFRβ with the ctβ antibody (third panel) or α-tubulin (bottom). (b) Quantification of PDGFRβ distribution upon inhibition of receptor tyrosine kinase activity with AG1296. Immunoblots were quantified as described in Fig. 1 . Error bars indicate SD. Quantification was based on four independent experiments. (c) Cells were stimulated with PDGF-BB or phosphatase activity was inhibited with sodium pervanadate, and lysates were immunoblotted with total PDGFR antibody (Y92) or lamin A/C for nuclear fractions (top two panels) or total PDGFRβ, α-tubulin, and pPDGFRβ antibody for cytoplasmic fractions (three bottom panels). Molecular mass was measured in kilodaltons. IB, immunoblotting; IP, immunoprecipitation.

    Techniques Used: Inhibition, Translocation Assay, Activity Assay, Immunoprecipitation, Western Blot

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    Article Title: New Approaches to Prevent LEOPARD Syndrome-associated Cardiac Hypertrophy by Specifically Targeting Shp2-dependent Signaling *
    Article Snippet: .. To induce strong tyrosine phosphorylation of focal adhesion kinase, NRCM were stimulated for 30 min with 10% fetal bovine serum in the presence of 0.1 m m pervanadate (generated by adding hydrogen peroxide to sodium orthovanadate stock solution (Sigma-Aldrich)). ..

    other:

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    Quantitative RT-PCR:

    Article Title: O-GlcNAcylation of SKN-1 modulates the lifespan and oxidative stress resistance in Caenorhabditis elegans
    Article Snippet: .. Oxidative stress resistance assay To examine the accumulation of SKN-1B/C::GFP in the intestinal nuclei and SKN-1 target gene expressions by RT-qPCR under oxidative stress, young adults were exposed to 5 mM t-butyl hydrogen peroxide tBHP (Sigma) for 90 minutes. ..

    Blocking Assay:

    Article Title: PDGFRβ translocates to the nucleus and regulates chromatin remodeling via TATA element–modifying factor 1
    Article Snippet: .. To enhance receptor phosphorylation in the absence of PDGF-BB stimulation, cells were treated with sodium pervanadate (1 mM hydrogen peroxide and 0.1 mM vanadate, freshly mixed) for 30 min. To block Golgi to ER retrograde traffic, cells were pretreated with 500 ng/ml brefeldin A (Sigma) for 1.5 h before PDGF-BB stimulation. .. To block lysosomal or proteasomal degradation, cells were pretreated for 3 h with 100 µM chloroquine or 25 µM MG132, respectively.

    Flow Cytometry:

    Article Title: Identification of 5-HT receptor subtypes enhancing inhibitory transmission in the rat spinal dorsal horn in vitro
    Article Snippet: .. The time necessary for the solution to flow from the stopcock to the surface of the spinal cord was approximately 5 s and the solution in the recording chamber was completely exchanged with a drug containing solution within 15 s. The drugs used were 5-HT hydrogen maleate (Sigma, St. Louis, MO, USA), tetrodotoxin (TTX) (Wako, Osaka, Japan), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (Sigma), DL-2-amino-5-phosphonovaleric acid (APV) (Sigma), BaCl2 (Sigma), strychnine (Sigma), bicuculline (Sigma), 4-bromo-3,6-dimethoxybenzocyclobuten-1-yl) methylamine hydrobromide (TBC-2) (Tocris Cookson, Bristol, UK), ketanserin (Sigma), α-methyl-5-(2-thienylmethoxy)-H-indole-3-ethanamine hydrochloride (BW723C86) (Sigma), 8,9-dchloro-2,3,4 4a-tetrahydro-1H-pyrazino[1,2-a] quinoxalin-5(6H)-one hydrochloride (WAY161503) (Tocris), 8-chloro-11-(1-piperazinyl)-5H-dibenzo[b,e][1,4]diazipine (N-desmethylclozpine) (Tocris), 1-(m-chlorophenyl)-biguanide (mCPBG) (Sigma), 3-tropanylindole-3-carboxylate methiodide (ICS-205,930) (Sigma), (±)-8-hydroxy-2-dipropylaminotetralin hydrobromide (8-OH-DPAT) (Sigma). ..

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    Millipore np 40 containing lysis buffer
    Np 40 Containing Lysis 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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    Millipore sodium pervanadate
    Sodium Pervanadate, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sodium pervanadate/product/Millipore
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
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