nonidet p 40 lysis buffer  (Thermo Fisher)


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  • 99
    Name:
    Glycerol
    Description:
    Thermo Scientific Pierce Glycerol is high purity glycerol greater than 99 pure for use in molecular biology methods
    Catalog Number:
    17904
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis
    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher nonidet p 40 lysis buffer
    ( A ). ( B ) Cdc25 activity during Fas-induced apoptosis. Samples from control extract (−α Fas), apoptotic extract (+α Fas), and nocodazole arrested extract (N-arrested, ≈50% in mitosis) were analyzed with anti-cdc25 immunoblotting ( Upper ). The relative cdc25 activities are shown graphically ( Lower ). ( C ) Anti-Wee1 blot of the extracts. ( D ) Wee1 activity during apoptosis. ( Middle  and  Bottom ) Western blot analysis of the active cyclin B/cdc2 beads treated with different extracts. ( Top ) H1 kinase activity of the same corresponding beads. Each extract (≈5 mg/ml total proteins) made in Nonidet P-40 lysis buffer was analyzed at three different dilutions: 1, 1/5, and 1/25. Five milligrams per milliliters BSA was used as a negative control.
    Thermo Scientific Pierce Glycerol is high purity glycerol greater than 99 pure for use in molecular biology methods
    https://www.bioz.com/result/nonidet p 40 lysis buffer/product/Thermo Fisher
    Average 99 stars, based on 45 article reviews
    Price from $9.99 to $1999.99
    nonidet p 40 lysis buffer - by Bioz Stars, 2020-11
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    Images

    1) Product Images from "Caspase-dependent activation of cyclin-dependent kinases during Fas-induced apoptosis in Jurkat cells"

    Article Title: Caspase-dependent activation of cyclin-dependent kinases during Fas-induced apoptosis in Jurkat cells

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    ( A ). ( B ) Cdc25 activity during Fas-induced apoptosis. Samples from control extract (−α Fas), apoptotic extract (+α Fas), and nocodazole arrested extract (N-arrested, ≈50% in mitosis) were analyzed with anti-cdc25 immunoblotting ( Upper ). The relative cdc25 activities are shown graphically ( Lower ). ( C ) Anti-Wee1 blot of the extracts. ( D ) Wee1 activity during apoptosis. ( Middle  and  Bottom ) Western blot analysis of the active cyclin B/cdc2 beads treated with different extracts. ( Top ) H1 kinase activity of the same corresponding beads. Each extract (≈5 mg/ml total proteins) made in Nonidet P-40 lysis buffer was analyzed at three different dilutions: 1, 1/5, and 1/25. Five milligrams per milliliters BSA was used as a negative control.
    Figure Legend Snippet: ( A ). ( B ) Cdc25 activity during Fas-induced apoptosis. Samples from control extract (−α Fas), apoptotic extract (+α Fas), and nocodazole arrested extract (N-arrested, ≈50% in mitosis) were analyzed with anti-cdc25 immunoblotting ( Upper ). The relative cdc25 activities are shown graphically ( Lower ). ( C ) Anti-Wee1 blot of the extracts. ( D ) Wee1 activity during apoptosis. ( Middle and Bottom ) Western blot analysis of the active cyclin B/cdc2 beads treated with different extracts. ( Top ) H1 kinase activity of the same corresponding beads. Each extract (≈5 mg/ml total proteins) made in Nonidet P-40 lysis buffer was analyzed at three different dilutions: 1, 1/5, and 1/25. Five milligrams per milliliters BSA was used as a negative control.

    Techniques Used: Activity Assay, Western Blot, Lysis, Negative Control

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    Article Title: MYC is a critical target of FBXW7
    Article Snippet: .. Production of lentivirus and shRNA Glycerol stocks of lentiviral shRNA constructs (pGIPZ, pTRIPZ) were obtained from Thermo Scientific Open Biosystems (Waltham, Massachusetts, USA) and grown in LB medium with 100цg/ml carbenicillin (Sigma) and 25цg/ml Zeocin (InvivoGen, San Diego, CA, USA). .. Clones used were FBXW7-7 (V2LHS_202932), FBXW7-8 (V2THS_89328), FBXW7-10 (V2THS_203045), UBE2I-3 (V2LHS_171776), and UBE2I-6 (V3LHS_376933).

    Fluorescence:

    Article Title: Frequent exchange of the DNA polymerase during bacterial chromosome replication
    Article Snippet: .. Fluorescence recovery after photobleaching (FRAP) Cells were grown in M9-Glycerol at 30°C, treated with cephalexin for 2 hr, harvested at early log-phase (OD600 0.1–0.2), concentrated and spotted onto a pad of 1% agarose in M9-Glycerol, contained in a gene frame (Thermo Scientific). ..

    Construct:

    Article Title: MYC is a critical target of FBXW7
    Article Snippet: .. Production of lentivirus and shRNA Glycerol stocks of lentiviral shRNA constructs (pGIPZ, pTRIPZ) were obtained from Thermo Scientific Open Biosystems (Waltham, Massachusetts, USA) and grown in LB medium with 100цg/ml carbenicillin (Sigma) and 25цg/ml Zeocin (InvivoGen, San Diego, CA, USA). .. Clones used were FBXW7-7 (V2LHS_202932), FBXW7-8 (V2THS_89328), FBXW7-10 (V2THS_203045), UBE2I-3 (V2LHS_171776), and UBE2I-6 (V3LHS_376933).

    Article Title: Crystal structure of DRIK1, a stress-responsive receptor-like pseudokinase, reveals the molecular basis for the absence of ATP binding
    Article Snippet: .. One micromolar of each purified construct of Zm DRIK1-KD was mixed with DSF buffer (100 mM K-phosphate, 150 mM NaCl, 10% glycerol, and pH 7.5) containing 1:1000 SYPRO Orange Protein Thermal Shift Dye (Life Technologies Corporation, Eugene, USA) in a 384-well plate containing 10 μM of each library compound. ..

    Purification:

    Article Title: Crystal structure of DRIK1, a stress-responsive receptor-like pseudokinase, reveals the molecular basis for the absence of ATP binding
    Article Snippet: .. One micromolar of each purified construct of Zm DRIK1-KD was mixed with DSF buffer (100 mM K-phosphate, 150 mM NaCl, 10% glycerol, and pH 7.5) containing 1:1000 SYPRO Orange Protein Thermal Shift Dye (Life Technologies Corporation, Eugene, USA) in a 384-well plate containing 10 μM of each library compound. ..

    Immunoprecipitation:

    Article Title: Acetylation of VGLL4 Regulates Hippo-YAP Signaling and Postnatal Cardiac Growth
    Article Snippet: .. The protein solution was diluted with 1 volume of immunoprecipitation buffer (lysis buffer without glycerol) and pre-cleared with protein A Dynabeads (Life Technologies, 10008D). ..

    Incubation:

    Article Title: Translesion synthesis mechanisms depend on the nature of DNA damage in UV-irradiated human cells
    Article Snippet: .. For experiments with the ssDNA-specific S1 endonuclease, after an IdU pulse, the cells were treated with CSK100 buffer (100 mM NaCl, 300 mM sucrose, 3 mM MgCl2, 10 mM MOPS, 0.5% Triton X-100) for 10 min at RT, then incubated with S1 nuclease buffer (50 mM NaCl, 30 mM sodium acetate pH 4.6, 10 mM zinc acetate and 5% glycerol) with or without S1 nuclease (Invitrogen, Life Technologies) at 20 U/ml for 30 min at 37ºC. ..

    Lysis:

    Article Title: Acetylation of VGLL4 Regulates Hippo-YAP Signaling and Postnatal Cardiac Growth
    Article Snippet: .. The protein solution was diluted with 1 volume of immunoprecipitation buffer (lysis buffer without glycerol) and pre-cleared with protein A Dynabeads (Life Technologies, 10008D). ..

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    Thermo Fisher immunoprecipitation ip lysis buffer
    Interactions of FIGLA, LHX8 and SOHLH1. ( A ) Representative protein domains (bHLH, LIM, homeobox) of FIGLA, SOHLH1 and LHX8. ( B ) FIGLA HA and LHX8 FLAG expression vectors were co-transfected into HEK-293T cells. Cell lysates were probed with HA and FLAG antibodies to detect input protein FIGLA and LHX8, respectively. After <t>immunoprecipitation</t> with HA and FLAG antibody, immunoblots were performed to detect FIGLA and associated LHX8 protein or LHX8 and associated FIGLA protein, respectively. ( C ) Same as (B) except that FIGLA HA and SOHLH1 MYC were co-transfected into HEK-293T cells. Antibodies to HA and MYC were used to detect input proteins and immunoprecipitate FIGLA and SOHLH1, respectively, and their associated proteins. ( D ) Same as (B) except that LHX8 FLAG and SOHLH1 MYC were co-transfected into HEK-293T cells. Antibodies to FLAG and MYC were used to detect input proteins and immunoprecipitate LHX8 and SOHLH1, respectively, and their associated proteins. ( E ) Co-expression of FIGLA, LHX8 and SOHLH1 in P0 ovaries from Figla FLAG mice. The dashed circles indicate co-expression of FIGLA, LHX8 and SOHLH1 in the same oocytes. Scale bar, 20 μm. ( F ) FIGLA, LHX8 and SOHLH1 appear to form a nuclear complex in oocytes. Representative of n = 3 (B–E) independent biological replicates with similar results per condition.
    Immunoprecipitation Ip Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation ip lysis buffer/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitation ip lysis buffer - by Bioz Stars, 2020-11
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    99
    Thermo Fisher np 40 buffer
    DDC-induced ROS formation is associated with a depletion of energy metabolism and antioxidant enzymes and is regulated by GAPDH. (A) Hepatocytes from C3H and C57BL mice were left untreated (−) or treated with either 0.1% DMSO vehicle (0) or the indicated concentrations of DDC for 48 h. Equal protein amounts <t>(NP-40</t> lysates) were analyzed for expression of the indicated proteins. Samples from each strain were analyzed on separate gels. PDIA4 levels were unaltered (loading control). (B) C57BL hepatocytes were transfected with Control or GAPDH siRNA for 24 h and were then cultured for an additional 24 h in the presence of 0.025% DMSO vehicle (0) or 100 µM DDC or were left untreated (−). The NP-40 cell lysates were analyzed for the expression of the various proteins indicated. (C) Overexpression of Flag-tagged mouse GAPDH in isolated hepatocytes. NP-40 lysates were analyzed for expression of Flag-tagged GAPDH and total GAPDH. The Coomassie stain is included as a loading control. (D) Flag-GAPDH expression in C57BL hepatocytes, as determined by immunostaining with a mouse anti-Flag antibody (representing overexpressed GAPDH; green) and a rabbit anti-GAPDH antibody (representing total GAPDH; red). (E) C57BL hepatocytes were mock transfected (Control) or transfected with GAPDH siRNA or Flag-GAPDH mouse cDNA for 24 h and were then treated with 100 µM DDC for an additional 24 h. Representative images of ROS signal (green) with DAPI nuclear counterstain (blue) are shown. ROS levels (quantified as described in Materials and methods) exhibited a 2.2-fold increase after GAPDH knockdown and a 10-fold decrease after GAPDH overexpression relative to control. Bars, 20 µm.
    Np 40 Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher pierce co immunoprecipitation lysis buffer
    SDS-PAGE Western blot analysis with MAb MUC16CT 2C6 . ( A ) OVCAR3 cell lysate blot (50 μg) probed with both MAbs M11 to the MUC16 ectodomain and the MUC16CT 2C6 to the cytoplasmic tail. The blot was first probed with the MUC16CT 2C6 antibody, then reprobed with the M11 antibody to confirm detection of MUC16. Note that the CT antibody weakly recognized the full length MUC16 and that M11 does not recognize cleaved MUC16 CT. ( B ) Western blot of MAb MUC16CT 2C6 immunoprecipitates from OVCAR3 cell lysates after preincubation of the cells with (+) or without (–) rZmpC to remove the MUC16 ectodomain. After <t>immunoprecipitation,</t> MAb MUC16CT 2C6 detected the CT domain in samples pretreated with rZmpC. We note variation in apparent contaminating high molecular weight bands in the IP most notably in the 4 H 60 min experiment with ZmpC, but the banding patterns are similar in all ZmpC treated samples at 30 and 60 min with increased exposure. The lack of detection of the full length MUC16 is likely due to its large size and potential removal from shearing during immunoprecipitation or lack of access of the mAb to the CT from the large MUC16 ectodomain. Without ZmpC treatment (–), the CT was weakly detected, due perhaps to lower protein loading (approximately 20 μg) than in A (50 μg) or to steric hindrance of access of the MAb MUC16CT 2C6 to the CT domain by the massive full length MUC16 molecule.
    Pierce Co Immunoprecipitation Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Interactions of FIGLA, LHX8 and SOHLH1. ( A ) Representative protein domains (bHLH, LIM, homeobox) of FIGLA, SOHLH1 and LHX8. ( B ) FIGLA HA and LHX8 FLAG expression vectors were co-transfected into HEK-293T cells. Cell lysates were probed with HA and FLAG antibodies to detect input protein FIGLA and LHX8, respectively. After immunoprecipitation with HA and FLAG antibody, immunoblots were performed to detect FIGLA and associated LHX8 protein or LHX8 and associated FIGLA protein, respectively. ( C ) Same as (B) except that FIGLA HA and SOHLH1 MYC were co-transfected into HEK-293T cells. Antibodies to HA and MYC were used to detect input proteins and immunoprecipitate FIGLA and SOHLH1, respectively, and their associated proteins. ( D ) Same as (B) except that LHX8 FLAG and SOHLH1 MYC were co-transfected into HEK-293T cells. Antibodies to FLAG and MYC were used to detect input proteins and immunoprecipitate LHX8 and SOHLH1, respectively, and their associated proteins. ( E ) Co-expression of FIGLA, LHX8 and SOHLH1 in P0 ovaries from Figla FLAG mice. The dashed circles indicate co-expression of FIGLA, LHX8 and SOHLH1 in the same oocytes. Scale bar, 20 μm. ( F ) FIGLA, LHX8 and SOHLH1 appear to form a nuclear complex in oocytes. Representative of n = 3 (B–E) independent biological replicates with similar results per condition.

    Journal: Nucleic Acids Research

    Article Title: FIGLA, LHX8 and SOHLH1 transcription factor networks regulate mouse oocyte growth and differentiation

    doi: 10.1093/nar/gkaa101

    Figure Lengend Snippet: Interactions of FIGLA, LHX8 and SOHLH1. ( A ) Representative protein domains (bHLH, LIM, homeobox) of FIGLA, SOHLH1 and LHX8. ( B ) FIGLA HA and LHX8 FLAG expression vectors were co-transfected into HEK-293T cells. Cell lysates were probed with HA and FLAG antibodies to detect input protein FIGLA and LHX8, respectively. After immunoprecipitation with HA and FLAG antibody, immunoblots were performed to detect FIGLA and associated LHX8 protein or LHX8 and associated FIGLA protein, respectively. ( C ) Same as (B) except that FIGLA HA and SOHLH1 MYC were co-transfected into HEK-293T cells. Antibodies to HA and MYC were used to detect input proteins and immunoprecipitate FIGLA and SOHLH1, respectively, and their associated proteins. ( D ) Same as (B) except that LHX8 FLAG and SOHLH1 MYC were co-transfected into HEK-293T cells. Antibodies to FLAG and MYC were used to detect input proteins and immunoprecipitate LHX8 and SOHLH1, respectively, and their associated proteins. ( E ) Co-expression of FIGLA, LHX8 and SOHLH1 in P0 ovaries from Figla FLAG mice. The dashed circles indicate co-expression of FIGLA, LHX8 and SOHLH1 in the same oocytes. Scale bar, 20 μm. ( F ) FIGLA, LHX8 and SOHLH1 appear to form a nuclear complex in oocytes. Representative of n = 3 (B–E) independent biological replicates with similar results per condition.

    Article Snippet: Immunoprecipitation and co-IPHarvested cells were lysed in immunoprecipitation (IP) lysis buffer (50 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40, 1 mM EDTA) supplemented with 1× protease inhibitor cocktail (Thermo Fisher Scientific) on ice for 30 min.

    Techniques: Expressing, Transfection, Immunoprecipitation, Western Blot, Mouse Assay

    DDC-induced ROS formation is associated with a depletion of energy metabolism and antioxidant enzymes and is regulated by GAPDH. (A) Hepatocytes from C3H and C57BL mice were left untreated (−) or treated with either 0.1% DMSO vehicle (0) or the indicated concentrations of DDC for 48 h. Equal protein amounts (NP-40 lysates) were analyzed for expression of the indicated proteins. Samples from each strain were analyzed on separate gels. PDIA4 levels were unaltered (loading control). (B) C57BL hepatocytes were transfected with Control or GAPDH siRNA for 24 h and were then cultured for an additional 24 h in the presence of 0.025% DMSO vehicle (0) or 100 µM DDC or were left untreated (−). The NP-40 cell lysates were analyzed for the expression of the various proteins indicated. (C) Overexpression of Flag-tagged mouse GAPDH in isolated hepatocytes. NP-40 lysates were analyzed for expression of Flag-tagged GAPDH and total GAPDH. The Coomassie stain is included as a loading control. (D) Flag-GAPDH expression in C57BL hepatocytes, as determined by immunostaining with a mouse anti-Flag antibody (representing overexpressed GAPDH; green) and a rabbit anti-GAPDH antibody (representing total GAPDH; red). (E) C57BL hepatocytes were mock transfected (Control) or transfected with GAPDH siRNA or Flag-GAPDH mouse cDNA for 24 h and were then treated with 100 µM DDC for an additional 24 h. Representative images of ROS signal (green) with DAPI nuclear counterstain (blue) are shown. ROS levels (quantified as described in Materials and methods) exhibited a 2.2-fold increase after GAPDH knockdown and a 10-fold decrease after GAPDH overexpression relative to control. Bars, 20 µm.

    Journal: The Journal of Cell Biology

    Article Title: Energy determinants GAPDH and NDPK act as genetic modifiers for hepatocyte inclusion formation

    doi: 10.1083/jcb.201102142

    Figure Lengend Snippet: DDC-induced ROS formation is associated with a depletion of energy metabolism and antioxidant enzymes and is regulated by GAPDH. (A) Hepatocytes from C3H and C57BL mice were left untreated (−) or treated with either 0.1% DMSO vehicle (0) or the indicated concentrations of DDC for 48 h. Equal protein amounts (NP-40 lysates) were analyzed for expression of the indicated proteins. Samples from each strain were analyzed on separate gels. PDIA4 levels were unaltered (loading control). (B) C57BL hepatocytes were transfected with Control or GAPDH siRNA for 24 h and were then cultured for an additional 24 h in the presence of 0.025% DMSO vehicle (0) or 100 µM DDC or were left untreated (−). The NP-40 cell lysates were analyzed for the expression of the various proteins indicated. (C) Overexpression of Flag-tagged mouse GAPDH in isolated hepatocytes. NP-40 lysates were analyzed for expression of Flag-tagged GAPDH and total GAPDH. The Coomassie stain is included as a loading control. (D) Flag-GAPDH expression in C57BL hepatocytes, as determined by immunostaining with a mouse anti-Flag antibody (representing overexpressed GAPDH; green) and a rabbit anti-GAPDH antibody (representing total GAPDH; red). (E) C57BL hepatocytes were mock transfected (Control) or transfected with GAPDH siRNA or Flag-GAPDH mouse cDNA for 24 h and were then treated with 100 µM DDC for an additional 24 h. Representative images of ROS signal (green) with DAPI nuclear counterstain (blue) are shown. ROS levels (quantified as described in Materials and methods) exhibited a 2.2-fold increase after GAPDH knockdown and a 10-fold decrease after GAPDH overexpression relative to control. Bars, 20 µm.

    Article Snippet: Isolated hepatocytes were either lysed in NP-40 buffer or subjected to subcellular fractionation using the NE-PER cytoplasmic/nuclear fractionation kit (Thermo Fisher Scientific) to obtain total, cytoplasmic, and nuclear-enriched fractions.

    Techniques: Mouse Assay, Expressing, Transfection, Cell Culture, Over Expression, Isolation, Staining, Immunostaining

    SDS-PAGE Western blot analysis with MAb MUC16CT 2C6 . ( A ) OVCAR3 cell lysate blot (50 μg) probed with both MAbs M11 to the MUC16 ectodomain and the MUC16CT 2C6 to the cytoplasmic tail. The blot was first probed with the MUC16CT 2C6 antibody, then reprobed with the M11 antibody to confirm detection of MUC16. Note that the CT antibody weakly recognized the full length MUC16 and that M11 does not recognize cleaved MUC16 CT. ( B ) Western blot of MAb MUC16CT 2C6 immunoprecipitates from OVCAR3 cell lysates after preincubation of the cells with (+) or without (–) rZmpC to remove the MUC16 ectodomain. After immunoprecipitation, MAb MUC16CT 2C6 detected the CT domain in samples pretreated with rZmpC. We note variation in apparent contaminating high molecular weight bands in the IP most notably in the 4 H 60 min experiment with ZmpC, but the banding patterns are similar in all ZmpC treated samples at 30 and 60 min with increased exposure. The lack of detection of the full length MUC16 is likely due to its large size and potential removal from shearing during immunoprecipitation or lack of access of the mAb to the CT from the large MUC16 ectodomain. Without ZmpC treatment (–), the CT was weakly detected, due perhaps to lower protein loading (approximately 20 μg) than in A (50 μg) or to steric hindrance of access of the MAb MUC16CT 2C6 to the CT domain by the massive full length MUC16 molecule.

    Journal: Glycobiology

    Article Title: Generation and characterization of a monoclonal antibody to the cytoplasmic tail of MUC16

    doi: 10.1093/glycob/cwx054

    Figure Lengend Snippet: SDS-PAGE Western blot analysis with MAb MUC16CT 2C6 . ( A ) OVCAR3 cell lysate blot (50 μg) probed with both MAbs M11 to the MUC16 ectodomain and the MUC16CT 2C6 to the cytoplasmic tail. The blot was first probed with the MUC16CT 2C6 antibody, then reprobed with the M11 antibody to confirm detection of MUC16. Note that the CT antibody weakly recognized the full length MUC16 and that M11 does not recognize cleaved MUC16 CT. ( B ) Western blot of MAb MUC16CT 2C6 immunoprecipitates from OVCAR3 cell lysates after preincubation of the cells with (+) or without (–) rZmpC to remove the MUC16 ectodomain. After immunoprecipitation, MAb MUC16CT 2C6 detected the CT domain in samples pretreated with rZmpC. We note variation in apparent contaminating high molecular weight bands in the IP most notably in the 4 H 60 min experiment with ZmpC, but the banding patterns are similar in all ZmpC treated samples at 30 and 60 min with increased exposure. The lack of detection of the full length MUC16 is likely due to its large size and potential removal from shearing during immunoprecipitation or lack of access of the mAb to the CT from the large MUC16 ectodomain. Without ZmpC treatment (–), the CT was weakly detected, due perhaps to lower protein loading (approximately 20 μg) than in A (50 μg) or to steric hindrance of access of the MAb MUC16CT 2C6 to the CT domain by the massive full length MUC16 molecule.

    Article Snippet: Cells were then washed once with Modified Dulbecco's PBS and lysed with ice-cold Pierce Co-Immunoprecipitation lysis buffer (0.025 M Tris, 0.15 M NaCl, 0.001 M EDTA, 1% NP-40, 5% glycerol; pH 7.4) plus Halt protease and phosphatase inhibitor cocktail (Thermo Scientific) and incubated on ice for 5 min with periodic mixing.

    Techniques: SDS Page, Western Blot, Immunoprecipitation, Molecular Weight