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  • 99
    Millipore pmsf
    Trypsin digestion generates a 69-kDa Ku86 protein from a full-length cloned human MM Ku86 protein (rhKu86) in vitro . Full-length rhKu86 was first expressed and purified from COS cells. Serial digestion (A) of rhKu86 (6.5 μg/sample) by trypsin (0.065 μg of Trypsin Gold/reaction up to 20 mins) was then performed using a limited proteolysis protocol from the manufacturer. The reactions were stopped by rapid cooling on ice; and the products of trypsin digestion were resolved by SDS PAGE and immunoblotted S10B1 anti-Ku86 mAb. The effect of protease inhibitors <t>aprotinin</t> (2.0 μg/mL final concentration) plus <t>PMSF</t> (100 μg/mL final concentration) on trypsin digestion was also determined on a larger sample of rhKu86 (13.0 μg/sample). Trypsin digestion (0.065 μg of Trypsin Gold/reaction for 1 min) of rhKu86 was performed as above either without (lane 1) or with (lane 2) protease inhibitors (B).
    Pmsf, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 32226 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmsf/product/Millipore
    Average 99 stars, based on 32226 article reviews
    Price from $9.99 to $1999.99
    pmsf - by Bioz Stars, 2020-08
    99/100 stars
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    99
    Cell Signaling Technology Inc pmsf
    Trypsin digestion generates a 69-kDa Ku86 protein from a full-length cloned human MM Ku86 protein (rhKu86) in vitro . Full-length rhKu86 was first expressed and purified from COS cells. Serial digestion (A) of rhKu86 (6.5 μg/sample) by trypsin (0.065 μg of Trypsin Gold/reaction up to 20 mins) was then performed using a limited proteolysis protocol from the manufacturer. The reactions were stopped by rapid cooling on ice; and the products of trypsin digestion were resolved by SDS PAGE and immunoblotted S10B1 anti-Ku86 mAb. The effect of protease inhibitors <t>aprotinin</t> (2.0 μg/mL final concentration) plus <t>PMSF</t> (100 μg/mL final concentration) on trypsin digestion was also determined on a larger sample of rhKu86 (13.0 μg/sample). Trypsin digestion (0.065 μg of Trypsin Gold/reaction for 1 min) of rhKu86 was performed as above either without (lane 1) or with (lane 2) protease inhibitors (B).
    Pmsf, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 952 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmsf/product/Cell Signaling Technology Inc
    Average 99 stars, based on 952 article reviews
    Price from $9.99 to $1999.99
    pmsf - by Bioz Stars, 2020-08
    99/100 stars
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    pmsf  (Abcam)
    94
    Abcam pmsf
    Trypsin digestion generates a 69-kDa Ku86 protein from a full-length cloned human MM Ku86 protein (rhKu86) in vitro . Full-length rhKu86 was first expressed and purified from COS cells. Serial digestion (A) of rhKu86 (6.5 μg/sample) by trypsin (0.065 μg of Trypsin Gold/reaction up to 20 mins) was then performed using a limited proteolysis protocol from the manufacturer. The reactions were stopped by rapid cooling on ice; and the products of trypsin digestion were resolved by SDS PAGE and immunoblotted S10B1 anti-Ku86 mAb. The effect of protease inhibitors <t>aprotinin</t> (2.0 μg/mL final concentration) plus <t>PMSF</t> (100 μg/mL final concentration) on trypsin digestion was also determined on a larger sample of rhKu86 (13.0 μg/sample). Trypsin digestion (0.065 μg of Trypsin Gold/reaction for 1 min) of rhKu86 was performed as above either without (lane 1) or with (lane 2) protease inhibitors (B).
    Pmsf, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmsf/product/Abcam
    Average 94 stars, based on 163 article reviews
    Price from $9.99 to $1999.99
    pmsf - by Bioz Stars, 2020-08
    94/100 stars
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    90
    Selleck Chemicals pmsf
    Trypsin digestion generates a 69-kDa Ku86 protein from a full-length cloned human MM Ku86 protein (rhKu86) in vitro . Full-length rhKu86 was first expressed and purified from COS cells. Serial digestion (A) of rhKu86 (6.5 μg/sample) by trypsin (0.065 μg of Trypsin Gold/reaction up to 20 mins) was then performed using a limited proteolysis protocol from the manufacturer. The reactions were stopped by rapid cooling on ice; and the products of trypsin digestion were resolved by SDS PAGE and immunoblotted S10B1 anti-Ku86 mAb. The effect of protease inhibitors <t>aprotinin</t> (2.0 μg/mL final concentration) plus <t>PMSF</t> (100 μg/mL final concentration) on trypsin digestion was also determined on a larger sample of rhKu86 (13.0 μg/sample). Trypsin digestion (0.065 μg of Trypsin Gold/reaction for 1 min) of rhKu86 was performed as above either without (lane 1) or with (lane 2) protease inhibitors (B).
    Pmsf, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 90/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmsf/product/Selleck Chemicals
    Average 90 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    pmsf - by Bioz Stars, 2020-08
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    pmsf  (Tocris)
    91
    Tocris pmsf
    Trypsin digestion generates a 69-kDa Ku86 protein from a full-length cloned human MM Ku86 protein (rhKu86) in vitro . Full-length rhKu86 was first expressed and purified from COS cells. Serial digestion (A) of rhKu86 (6.5 μg/sample) by trypsin (0.065 μg of Trypsin Gold/reaction up to 20 mins) was then performed using a limited proteolysis protocol from the manufacturer. The reactions were stopped by rapid cooling on ice; and the products of trypsin digestion were resolved by SDS PAGE and immunoblotted S10B1 anti-Ku86 mAb. The effect of protease inhibitors <t>aprotinin</t> (2.0 μg/mL final concentration) plus <t>PMSF</t> (100 μg/mL final concentration) on trypsin digestion was also determined on a larger sample of rhKu86 (13.0 μg/sample). Trypsin digestion (0.065 μg of Trypsin Gold/reaction for 1 min) of rhKu86 was performed as above either without (lane 1) or with (lane 2) protease inhibitors (B).
    Pmsf, supplied by Tocris, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmsf/product/Tocris
    Average 91 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    pmsf - by Bioz Stars, 2020-08
    91/100 stars
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    Image Search Results


    Trypsin digestion generates a 69-kDa Ku86 protein from a full-length cloned human MM Ku86 protein (rhKu86) in vitro . Full-length rhKu86 was first expressed and purified from COS cells. Serial digestion (A) of rhKu86 (6.5 μg/sample) by trypsin (0.065 μg of Trypsin Gold/reaction up to 20 mins) was then performed using a limited proteolysis protocol from the manufacturer. The reactions were stopped by rapid cooling on ice; and the products of trypsin digestion were resolved by SDS PAGE and immunoblotted S10B1 anti-Ku86 mAb. The effect of protease inhibitors aprotinin (2.0 μg/mL final concentration) plus PMSF (100 μg/mL final concentration) on trypsin digestion was also determined on a larger sample of rhKu86 (13.0 μg/sample). Trypsin digestion (0.065 μg of Trypsin Gold/reaction for 1 min) of rhKu86 was performed as above either without (lane 1) or with (lane 2) protease inhibitors (B).

    Journal: Cancer Cell International

    Article Title: Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells

    doi: 10.1186/1475-2867-8-4

    Figure Lengend Snippet: Trypsin digestion generates a 69-kDa Ku86 protein from a full-length cloned human MM Ku86 protein (rhKu86) in vitro . Full-length rhKu86 was first expressed and purified from COS cells. Serial digestion (A) of rhKu86 (6.5 μg/sample) by trypsin (0.065 μg of Trypsin Gold/reaction up to 20 mins) was then performed using a limited proteolysis protocol from the manufacturer. The reactions were stopped by rapid cooling on ice; and the products of trypsin digestion were resolved by SDS PAGE and immunoblotted S10B1 anti-Ku86 mAb. The effect of protease inhibitors aprotinin (2.0 μg/mL final concentration) plus PMSF (100 μg/mL final concentration) on trypsin digestion was also determined on a larger sample of rhKu86 (13.0 μg/sample). Trypsin digestion (0.065 μg of Trypsin Gold/reaction for 1 min) of rhKu86 was performed as above either without (lane 1) or with (lane 2) protease inhibitors (B).

    Article Snippet: Intracellular inhibition of protease digestion In order to inhibit intracellular protease activity, 5 × 106 cells/sample were treated with: Complete™ protease inhibitor tablets; either 1× (1 tablet for every 50 mL of media) or 2× (2 tablets for every 50 mL of media); or treated with antipain (2.0 μg/mL) plus leupeptin (2.0 μg/mL) (both from Sigma-Aldrich, St Louis, MO, USA) for cysteine protease inhibition; or aprotinin (2.0 μg/mL) plus PMSF (100 μg/mL) (both from Sigma-Aldrich) for serine protease inhibition; for to 24 hrs.

    Techniques: Clone Assay, In Vitro, Purification, SDS Page, Concentration Assay

    Proteolysis plays an important role in the intracellular generation of the 69-kDa Ku86v variant protein in MM cell lines . RPMI 8226 (A) and SGH-MM5 (data not shown) MM cell lines (5.0 × 10 6 cells/sample) were first incubated for 18 hrs with either 1× or 2× Complete™ protease inhibitor tablets (lanes 2 and 3); and then washed and checked for viability using trypan blue exclusion assay. Mock experiments in which cell lines were incubated for 18 hrs with media alone (lane 1) served as negative controls. Cell lysates were resolved by SDS-PAGE and immunoblotted using S10B1 anti-Ku86 mAb. The RPMI 8226 MM cell line s (B) was also incubated for 24 hrs with 2× Complete™ protease inhibitor tablets (lane 2), antipain (2.0 μg/mL final concentration) plus leupeptin (2.0 μg/mL final concentration) (lane 3), or aprotinin (2.0 μg/mL final concentration) plus PMSF (100 μg/mL final concentration) (lane 4) and analyzed in the same way as above. Mock experiments in which cell lines were incubated for 24 hrs with media alone (lane 1) again served as negative controls. Membranes were stripped and re-probed using anti-actin mAb (control) to confirm equal protein loading. Cell lysates were resolved by SDS-PAGE and immunoblotted using S10B1 anti-Ku86 mAb. Relative expression of 69-kDa Ku86v-DNA complexes (normalized to weakest band) was determined using image densitometry. All experiments were performed in triplicate.

    Journal: Cancer Cell International

    Article Title: Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells

    doi: 10.1186/1475-2867-8-4

    Figure Lengend Snippet: Proteolysis plays an important role in the intracellular generation of the 69-kDa Ku86v variant protein in MM cell lines . RPMI 8226 (A) and SGH-MM5 (data not shown) MM cell lines (5.0 × 10 6 cells/sample) were first incubated for 18 hrs with either 1× or 2× Complete™ protease inhibitor tablets (lanes 2 and 3); and then washed and checked for viability using trypan blue exclusion assay. Mock experiments in which cell lines were incubated for 18 hrs with media alone (lane 1) served as negative controls. Cell lysates were resolved by SDS-PAGE and immunoblotted using S10B1 anti-Ku86 mAb. The RPMI 8226 MM cell line s (B) was also incubated for 24 hrs with 2× Complete™ protease inhibitor tablets (lane 2), antipain (2.0 μg/mL final concentration) plus leupeptin (2.0 μg/mL final concentration) (lane 3), or aprotinin (2.0 μg/mL final concentration) plus PMSF (100 μg/mL final concentration) (lane 4) and analyzed in the same way as above. Mock experiments in which cell lines were incubated for 24 hrs with media alone (lane 1) again served as negative controls. Membranes were stripped and re-probed using anti-actin mAb (control) to confirm equal protein loading. Cell lysates were resolved by SDS-PAGE and immunoblotted using S10B1 anti-Ku86 mAb. Relative expression of 69-kDa Ku86v-DNA complexes (normalized to weakest band) was determined using image densitometry. All experiments were performed in triplicate.

    Article Snippet: Intracellular inhibition of protease digestion In order to inhibit intracellular protease activity, 5 × 106 cells/sample were treated with: Complete™ protease inhibitor tablets; either 1× (1 tablet for every 50 mL of media) or 2× (2 tablets for every 50 mL of media); or treated with antipain (2.0 μg/mL) plus leupeptin (2.0 μg/mL) (both from Sigma-Aldrich, St Louis, MO, USA) for cysteine protease inhibition; or aprotinin (2.0 μg/mL) plus PMSF (100 μg/mL) (both from Sigma-Aldrich) for serine protease inhibition; for to 24 hrs.

    Techniques: Variant Assay, Incubation, Protease Inhibitor, Trypan Blue Exclusion Assay, SDS Page, Concentration Assay, Expressing