monoclonal anti flag m2 peroxidase hrp  (Millipore)


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    Name:
    Monoclonal ANTI FLAG M2 Peroxidase HRP antibody
    Description:
    The Monoclonal ANTI FLAG M2 Peroxidase is a mouse IgG antibody covalently conjugated to horseradish peroxidase HRP The antibody binds to FLAG fusion proteins and recognizes the FLAG epitope at N terminal Met N terminal C terminal and internal FLAG peptides
    Catalog Number:
    a8592
    Price:
    None
    Applications:
    Suggested dilution for immunocytochemistry and western blotting 1:100 to 1:1000Suggested dilution for ELISA 1:20,000Browse additional application references in our FLAG(R) Literature portal.
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    Structured Review

    Millipore monoclonal anti flag m2 peroxidase hrp
    Effect of alkaline phosphatase fusion on performance in Western blot . scFv-APs demonstrate improved sensitivity in Western blot. Recombinant human desmin was detected with equimolar concentrations of anti-desmin scFv clone C10 (applied at a concentration of 2 × 10 -8 M) expressed as either scFv or scFv-AP formats. Detection was by <t>anti-FLAG</t> antibody coupled to horse-radish peroxidase and addition of ECL western-blotting substrate.
    The Monoclonal ANTI FLAG M2 Peroxidase is a mouse IgG antibody covalently conjugated to horseradish peroxidase HRP The antibody binds to FLAG fusion proteins and recognizes the FLAG epitope at N terminal Met N terminal C terminal and internal FLAG peptides
    https://www.bioz.com/result/monoclonal anti flag m2 peroxidase hrp/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti flag m2 peroxidase hrp - by Bioz Stars, 2020-04
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    Images

    1) Product Images from "A simple vector system to improve performance and utilisation of recombinant antibodies"

    Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies

    Journal: BMC Biotechnology

    doi: 10.1186/1472-6750-6-46

    Effect of alkaline phosphatase fusion on performance in Western blot . scFv-APs demonstrate improved sensitivity in Western blot. Recombinant human desmin was detected with equimolar concentrations of anti-desmin scFv clone C10 (applied at a concentration of 2 × 10 -8 M) expressed as either scFv or scFv-AP formats. Detection was by anti-FLAG antibody coupled to horse-radish peroxidase and addition of ECL western-blotting substrate.
    Figure Legend Snippet: Effect of alkaline phosphatase fusion on performance in Western blot . scFv-APs demonstrate improved sensitivity in Western blot. Recombinant human desmin was detected with equimolar concentrations of anti-desmin scFv clone C10 (applied at a concentration of 2 × 10 -8 M) expressed as either scFv or scFv-AP formats. Detection was by anti-FLAG antibody coupled to horse-radish peroxidase and addition of ECL western-blotting substrate.

    Techniques Used: Western Blot, Recombinant, Concentration Assay

    Effect of alkaline phosphatase fusion on performance in IHC. A. scFv-APs demonstrate improved sensitivity in IHC. HER2 clones are more sensitive when expressed in pSANG14-3F than pSANG10-3F Monovalent G98A, with an affinity of 320 nM, gave weakly positive staining of the antigen in breast cancer tissue. Whereas, bivalent G98A gave staining equivalent to the monomeric scFv clone C6.5, which has a 20-fold higher affinity. Bivalency does not visually improve C6.5 staining. Detection was by anti-FLAG antibody coupled to biotin with the addition of streptavidin-horse-radish peroxidase and a tyramine signal amplification step. B. Image analysis was used to quantify the staining intensities of all five scFvs as both monomeric scFv and bivalent scFv-AP.
    Figure Legend Snippet: Effect of alkaline phosphatase fusion on performance in IHC. A. scFv-APs demonstrate improved sensitivity in IHC. HER2 clones are more sensitive when expressed in pSANG14-3F than pSANG10-3F Monovalent G98A, with an affinity of 320 nM, gave weakly positive staining of the antigen in breast cancer tissue. Whereas, bivalent G98A gave staining equivalent to the monomeric scFv clone C6.5, which has a 20-fold higher affinity. Bivalency does not visually improve C6.5 staining. Detection was by anti-FLAG antibody coupled to biotin with the addition of streptavidin-horse-radish peroxidase and a tyramine signal amplification step. B. Image analysis was used to quantify the staining intensities of all five scFvs as both monomeric scFv and bivalent scFv-AP.

    Techniques Used: Immunohistochemistry, Clone Assay, Staining, Amplification

    Related Articles

    Stable Transfection:

    Article Title: Targeting scFv-Fc-scTRAIL fusion proteins to tumor cells
    Article Snippet: Materials Receptor-Fc and scFv-Fc fusion proteins were produced with stably transfected HEK293T cells and purified from the supernatant by protein A affinity chromatography as described previously [ , ]. .. HRP- and PE-conjugated anti-FLAG antibodies were obtained from Sigma-Aldrich (A8592) and Miltenyi Biotec (130-101-576), respectively.

    Synthesized:

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: Recombinant PP2B, the dimeric form, was synthesized in High Five cells by co-infection with the recombinant baculovirus encoding His8x -tagged hPP2B-Aα (vHis-PP2B-Aα) and FLAG-tagged hPP2B-Bα (vFLAG-PP2B- Bα). .. Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare).

    Article Title: Guanylate cyclase 2C agonism corrects CFTR mutants
    Article Snippet: Antibodies in this study included anti-CFTR antibodies R1104 (Eric Sorscher lab, CF Center, University of Alabama, Birmingham, Alabama, USA [presently, Emory University, Atlanta, Georgia, USA]), 24-1 (MAB25031, R & D Systems), and CF3 (ab2784, Abcam); anti-HA (3724, Cell Signaling Technologies); and anti–FLAG HRP (A8592, clone M2, MilliporeSigma). .. STc was custom synthesized from American Peptide Company.

    Construct:

    Article Title: Multiple layers of transcriptional regulation by PLZF in NKT-cell development
    Article Snippet: The following plasmids were used: pEF1a-BirA-V5his (a gift from K. Zhao, NIH, Bethesda), pCMV-HA-HDAC1 (a gift from J. Colgan, University of Iowa, Iowa City, IA), pcDNA3-Myc-CUL3 (a gift from Y. Xiong, University of North Carolina at Chapel Hill, Chapel Hill, NC), and pcDNA5-PLZF and pcDNA5-A/F/PLZF plasmids, constructed using standard molecular biology techniques. .. The following antibodies were used: anti-PLZF (AF2944; R & D Systems), anti-CUL3 (A301-109A; Bethyl Laboratories), anti-HDAC1 (ab-7028; Abcam), streptavidin-HRP (S2438; Sigma-Aldrich), anti–Flag-HRP (A8592; Sigma-Aldrich), anti-Bach2 (12809; CST), and anti-GAPDH (G9545; Sigma-Aldrich).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Functional competence of a partially engaged GPCR–β-arrestin complex
    Article Snippet: .. ELISA based assembly of β2 V2 R–βarr1-Fab30 complex For ELISA based in-vitro assembly of β2 V2 R–βarr1–Fab/ScFv30 complexes, purified Fab/ScFv30 (in 20 mM Hepes, pH 7.4, 100 mM NaCl) was first immobilized on 96 well MaxiSorp polystyrene plates (Nunc) at room temperature for 1 h. Afterwards, potential non-specific binding sites in the wells were blocked by incubation with 1% BSA at room temperature for 1 h. Subsequently, mixture of ligand stimulated cell lysate (or purified receptor) was added to the wells and incubated at room temperature for 1 h. Wells were washed extensively using 20 mM Hepes, pH 7.4, 100 mM NaCl, 0.01% MNG and then incubated with 1:2,000 dilution of HRP-coupled anti-FLAG M2 antibody (Sigma, catalog number A8592). .. After 1 h incubation, wells were extensively washed and assembly of the complex was visualized by adding 3,3′,5,5′-tetramethylbenzidine (TMB) ELISA (Genscript or Thermo).

    Incubation:

    Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies
    Article Snippet: .. Where anti-FLAG and ECL detection was required, the membranes were incubated with 0.22 μg/ml of monoclonal Anti-FLAG® M2 peroxidase (HRP) (Sigma-Aldrich, Poole, UK) in M-PBS-T for 1 h at room temperature on an orbital shaker, and ECL western blotting substrate added according to the supplier's instructions (Pierce, Cramlington, UK). .. The membranes were scanned on the Typhoon scanner and analysed using ImageQuant software.

    Article Title: A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation
    Article Snippet: .. The membrane was washed with TBST (three times, 25 ml) and incubated with α-FLAG-HRP (Sigma A-8592, diluted 1:3,000 in TBST) for 1 h. The membrane was washed with TBST (three times, 25 ml) and developed by using SuperSignal West Pico Chemiluminescent Substrate (Pierce). .. Lectin Staining of Cells. ldlD cells were incubated with acetylated sugars as described above and stained with FITC-labeled lectins (EY Laboratories): HPA (1 μg/ml) or jacalin (5 μg/ml) dissolved in PBS (pH 7.4) in the dark for 1 h at 4°C.

    Article Title: Functional competence of a partially engaged GPCR–β-arrestin complex
    Article Snippet: .. ELISA based assembly of β2 V2 R–βarr1-Fab30 complex For ELISA based in-vitro assembly of β2 V2 R–βarr1–Fab/ScFv30 complexes, purified Fab/ScFv30 (in 20 mM Hepes, pH 7.4, 100 mM NaCl) was first immobilized on 96 well MaxiSorp polystyrene plates (Nunc) at room temperature for 1 h. Afterwards, potential non-specific binding sites in the wells were blocked by incubation with 1% BSA at room temperature for 1 h. Subsequently, mixture of ligand stimulated cell lysate (or purified receptor) was added to the wells and incubated at room temperature for 1 h. Wells were washed extensively using 20 mM Hepes, pH 7.4, 100 mM NaCl, 0.01% MNG and then incubated with 1:2,000 dilution of HRP-coupled anti-FLAG M2 antibody (Sigma, catalog number A8592). .. After 1 h incubation, wells were extensively washed and assembly of the complex was visualized by adding 3,3′,5,5′-tetramethylbenzidine (TMB) ELISA (Genscript or Thermo).

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: The experimental conditions for rhPP2B dimer and rhPP2C production were same as those of the previous study , except for a change in the incubation temperature after the infection of High Five insect cells with recombinant baculoviruses. .. Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare).

    Article Title: Stimulator of interferon genes (STING) provides insect antiviral immunity by promoting Dredd caspase–mediated NF-κB activation
    Article Snippet: 20 μl of protein-agarose A/G beads (Santa Cruz Biotechnology) was added and incubated overnight. .. After transfer to a polyvinylidene difluoride membrane (GE Healthcare), samples were immunoblotted with anti-HA-HRP antibody (catalogue no. A00169-40, Genscript, Nanjing, China), anti-FLAG-HRP antibody (catalogue no. A8592-2MG, Sigma), or GAPDH antibody (catalogue no. abs830030, Shanghai, China) following the standard procedure and then developed using SuperSignalTM West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) as described previously ( ).

    Expressing:

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare). .. The expression level of His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα at 19 °C were similar to those at 27 °C (Supplementary Fig. S3 in the online version at DOI: 10.1016/j.btre.2016.07.004 ).

    BIA-KA:

    Article Title: Stimulator of interferon genes (STING) provides insect antiviral immunity by promoting Dredd caspase–mediated NF-κB activation
    Article Snippet: Protein concentration of whole-cell lysate was estimated by a BCA assay (Beyotime). .. After transfer to a polyvinylidene difluoride membrane (GE Healthcare), samples were immunoblotted with anti-HA-HRP antibody (catalogue no. A00169-40, Genscript, Nanjing, China), anti-FLAG-HRP antibody (catalogue no. A8592-2MG, Sigma), or GAPDH antibody (catalogue no. abs830030, Shanghai, China) following the standard procedure and then developed using SuperSignalTM West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) as described previously ( ).

    Western Blot:

    Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies
    Article Snippet: .. Where anti-FLAG and ECL detection was required, the membranes were incubated with 0.22 μg/ml of monoclonal Anti-FLAG® M2 peroxidase (HRP) (Sigma-Aldrich, Poole, UK) in M-PBS-T for 1 h at room temperature on an orbital shaker, and ECL western blotting substrate added according to the supplier's instructions (Pierce, Cramlington, UK). .. The membranes were scanned on the Typhoon scanner and analysed using ImageQuant software.

    Article Title: Multiple layers of transcriptional regulation by PLZF in NKT-cell development
    Article Snippet: Paragraph title: Immunoprecipitation and Western Blot Analysis. ... The following antibodies were used: anti-PLZF (AF2944; R & D Systems), anti-CUL3 (A301-109A; Bethyl Laboratories), anti-HDAC1 (ab-7028; Abcam), streptavidin-HRP (S2438; Sigma-Aldrich), anti–Flag-HRP (A8592; Sigma-Aldrich), anti-Bach2 (12809; CST), and anti-GAPDH (G9545; Sigma-Aldrich).

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: .. Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare). .. The expression level of His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα at 19 °C were similar to those at 27 °C (Supplementary Fig. S3 in the online version at DOI: 10.1016/j.btre.2016.07.004 ).

    Article Title: ABHD5 stimulates PNPLA1-mediated ω-O-acylceramide biosynthesis essential for a functional skin permeability barrier [S]
    Article Snippet: Immunoblot analyses were performed according to standard protocols using anti-FLAG M2-HRP (A8592; Sigma-Aldrich) antibody or anti-His (27-4710-01; GE Healthcare), anti-Xpress (R910-25; Invitrogen) or anti-GAPDH primary antibody (2118S; Cell Signaling Technology, Danvers, MA, USA), and the appropriate HRP-conjugated secondary antibody. .. HRP-conjugated IgG was detected using Clarity Western ECL Substrate (Bio-Rad).

    Article Title: Identification of the Mtus1 Splice Variant as a Novel Inhibitory Factor Against Cardiac Hypertrophy
    Article Snippet: Antibodies and Reagents Phenylephrine (PE; 046K1351), angiotensin II (Ang II; A9525), and anti‐FLAG M2 monoclonal antibody (A8592, F1804) were purchased from Sigma–Aldrich (St. Louis, MO). .. Antibodies against Actin (sc‐1615) and donkey anti‐goat IgG (sc‐2020) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA) for Western blotting.

    Article Title: Cardiac L-type calcium channel (Cav1.2) associates with ? subunits
    Article Snippet: Paragraph title: Immunoprecipitation and immunoblots ... The anti-α2 antibody (D219) and the anti-Flag antibody (A8592) were from Sigma.

    Transfection:

    Article Title: Targeting scFv-Fc-scTRAIL fusion proteins to tumor cells
    Article Snippet: Materials Receptor-Fc and scFv-Fc fusion proteins were produced with stably transfected HEK293T cells and purified from the supernatant by protein A affinity chromatography as described previously [ , ]. .. HRP- and PE-conjugated anti-FLAG antibodies were obtained from Sigma-Aldrich (A8592) and Miltenyi Biotec (130-101-576), respectively.

    Ligation:

    Article Title: A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation
    Article Snippet: Staudinger ligation was performed by incubation of 20 μl of the lysate (5 mg/ml protein) with 2 μl of phosphine-FLAG (5 mM in PBS, pH 7.4) for 6 h at 37°C. .. The membrane was washed with TBST (three times, 25 ml) and incubated with α-FLAG-HRP (Sigma A-8592, diluted 1:3,000 in TBST) for 1 h. The membrane was washed with TBST (three times, 25 ml) and developed by using SuperSignal West Pico Chemiluminescent Substrate (Pierce).

    Infection:

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: His8x -tagged hPP2Cα was expressed by seeding a spinner flask (1L) with High Five cells grown in Express Five serum-free medium at a density of 2 × 106 cells/ml and infecting them with the recombinant baculoviruses at a multiplicity of infection (m.o.i.) of 3. .. Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare).

    other:

    Article Title: SPOP regulates prostate epithelial cell proliferation and promotes ubiquitination and turnover of cMYC oncoprotein
    Article Snippet: Reagents and Antibodies The antibodies used in this study were mouse-monoclonal anti-FLAG-M2 (#F1804), mouse anti-β-Actin (#A2228), mouse anti-FLAG-HRP (#A8592), anti-rat IgG-HRP (#A9037 Sigma), (from Sigma, St Louis, MO); rabbit-polyclonal anti-AR(N-20) (#SC-816), mouse-monoclonal anti-SPOP(B-8) (#SC-377206), and goat-polyclonal anti-SPOP(C-14) (#SC-66649), goat anti-rabbit IgG-HRP (#SC-2004), and goat anti-mouse IgG-HRP (#SC-2005) (Santa Cruz Biotechnologies, Santa Cruz, CA); rat anti-HA-HRP (Roche); mouse anti-c-Myc-HRP (#MA1-81357, Thermo Scientific); rabbit monoclonal anti-c-Myc (#5605), mouse monoclonal anti-Cre Recombinase (#15036) (Cell Signaling, Danvers, MA); mouse-monoclonal anti-K8 (#MMS-162P), rabbit polyclonal anti-K5 (#PRB-160P) (Covance, Berkeley, CA).

    Droplet Countercurrent Chromatography:

    Article Title: A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation
    Article Snippet: Total protein concentration was determined by dextran-coated charcoal (DCC) assay (Bio-Rad) to be ≈5 mg/ml. .. The membrane was washed with TBST (three times, 25 ml) and incubated with α-FLAG-HRP (Sigma A-8592, diluted 1:3,000 in TBST) for 1 h. The membrane was washed with TBST (three times, 25 ml) and developed by using SuperSignal West Pico Chemiluminescent Substrate (Pierce).

    Protein Concentration:

    Article Title: A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation
    Article Snippet: Total protein concentration was determined by dextran-coated charcoal (DCC) assay (Bio-Rad) to be ≈5 mg/ml. .. The membrane was washed with TBST (three times, 25 ml) and incubated with α-FLAG-HRP (Sigma A-8592, diluted 1:3,000 in TBST) for 1 h. The membrane was washed with TBST (three times, 25 ml) and developed by using SuperSignal West Pico Chemiluminescent Substrate (Pierce).

    Article Title: Stimulator of interferon genes (STING) provides insect antiviral immunity by promoting Dredd caspase–mediated NF-κB activation
    Article Snippet: Protein concentration of whole-cell lysate was estimated by a BCA assay (Beyotime). .. After transfer to a polyvinylidene difluoride membrane (GE Healthcare), samples were immunoblotted with anti-HA-HRP antibody (catalogue no. A00169-40, Genscript, Nanjing, China), anti-FLAG-HRP antibody (catalogue no. A8592-2MG, Sigma), or GAPDH antibody (catalogue no. abs830030, Shanghai, China) following the standard procedure and then developed using SuperSignalTM West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) as described previously ( ).

    Binding Assay:

    Article Title: Functional competence of a partially engaged GPCR–β-arrestin complex
    Article Snippet: .. ELISA based assembly of β2 V2 R–βarr1-Fab30 complex For ELISA based in-vitro assembly of β2 V2 R–βarr1–Fab/ScFv30 complexes, purified Fab/ScFv30 (in 20 mM Hepes, pH 7.4, 100 mM NaCl) was first immobilized on 96 well MaxiSorp polystyrene plates (Nunc) at room temperature for 1 h. Afterwards, potential non-specific binding sites in the wells were blocked by incubation with 1% BSA at room temperature for 1 h. Subsequently, mixture of ligand stimulated cell lysate (or purified receptor) was added to the wells and incubated at room temperature for 1 h. Wells were washed extensively using 20 mM Hepes, pH 7.4, 100 mM NaCl, 0.01% MNG and then incubated with 1:2,000 dilution of HRP-coupled anti-FLAG M2 antibody (Sigma, catalog number A8592). .. After 1 h incubation, wells were extensively washed and assembly of the complex was visualized by adding 3,3′,5,5′-tetramethylbenzidine (TMB) ELISA (Genscript or Thermo).

    Purification:

    Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies
    Article Snippet: Where anti-FLAG and ECL detection was required, the membranes were incubated with 0.22 μg/ml of monoclonal Anti-FLAG® M2 peroxidase (HRP) (Sigma-Aldrich, Poole, UK) in M-PBS-T for 1 h at room temperature on an orbital shaker, and ECL western blotting substrate added according to the supplier's instructions (Pierce, Cramlington, UK). .. For alkaline phosphatase detection in western blot, scFv or scFv-AP, expressed from pSANG10-3F or pSANG14-3F respectively, purified or in diluted crude bacterial cell lysates were used as the primary antibody as described above.

    Article Title: Functional competence of a partially engaged GPCR–β-arrestin complex
    Article Snippet: .. ELISA based assembly of β2 V2 R–βarr1-Fab30 complex For ELISA based in-vitro assembly of β2 V2 R–βarr1–Fab/ScFv30 complexes, purified Fab/ScFv30 (in 20 mM Hepes, pH 7.4, 100 mM NaCl) was first immobilized on 96 well MaxiSorp polystyrene plates (Nunc) at room temperature for 1 h. Afterwards, potential non-specific binding sites in the wells were blocked by incubation with 1% BSA at room temperature for 1 h. Subsequently, mixture of ligand stimulated cell lysate (or purified receptor) was added to the wells and incubated at room temperature for 1 h. Wells were washed extensively using 20 mM Hepes, pH 7.4, 100 mM NaCl, 0.01% MNG and then incubated with 1:2,000 dilution of HRP-coupled anti-FLAG M2 antibody (Sigma, catalog number A8592). .. After 1 h incubation, wells were extensively washed and assembly of the complex was visualized by adding 3,3′,5,5′-tetramethylbenzidine (TMB) ELISA (Genscript or Thermo).

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare). .. The purified rhPP2B dimer and rhPP2Cα were subjected to 13.5% and 12% SDS-PAGE, respectively, following which they were visualized using Coomassie Brilliant Blue R staining ( ).

    Article Title: Targeting scFv-Fc-scTRAIL fusion proteins to tumor cells
    Article Snippet: Materials Receptor-Fc and scFv-Fc fusion proteins were produced with stably transfected HEK293T cells and purified from the supernatant by protein A affinity chromatography as described previously [ , ]. .. HRP- and PE-conjugated anti-FLAG antibodies were obtained from Sigma-Aldrich (A8592) and Miltenyi Biotec (130-101-576), respectively.

    Blocking Assay:

    Article Title: Cardiac L-type calcium channel (Cav1.2) associates with ? subunits
    Article Snippet: The anti-γ6 antibody (SC-67458; Santa Cruz Biotechnologies, Santa Cruz, CA, USA) and blocking peptide (SC-67458P; Santa Cruz Biotechnology) was used to probe heart and brain lysates (see above). .. The anti-α2 antibody (D219) and the anti-Flag antibody (A8592) were from Sigma.

    Staining:

    Article Title: A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation
    Article Snippet: The membrane was washed with TBST (three times, 25 ml) and incubated with α-FLAG-HRP (Sigma A-8592, diluted 1:3,000 in TBST) for 1 h. The membrane was washed with TBST (three times, 25 ml) and developed by using SuperSignal West Pico Chemiluminescent Substrate (Pierce). .. Lectin Staining of Cells. ldlD cells were incubated with acetylated sugars as described above and stained with FITC-labeled lectins (EY Laboratories): HPA (1 μg/ml) or jacalin (5 μg/ml) dissolved in PBS (pH 7.4) in the dark for 1 h at 4°C.

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare). .. The purified rhPP2B dimer and rhPP2Cα were subjected to 13.5% and 12% SDS-PAGE, respectively, following which they were visualized using Coomassie Brilliant Blue R staining ( ).

    Article Title: ABHD5 stimulates PNPLA1-mediated ω-O-acylceramide biosynthesis essential for a functional skin permeability barrier [S]
    Article Snippet: Immunoblot analyses were performed according to standard protocols using anti-FLAG M2-HRP (A8592; Sigma-Aldrich) antibody or anti-His (27-4710-01; GE Healthcare), anti-Xpress (R910-25; Invitrogen) or anti-GAPDH primary antibody (2118S; Cell Signaling Technology, Danvers, MA, USA), and the appropriate HRP-conjugated secondary antibody. .. Finally, membrane-bound proteins were stained with Coomassie blue staining solution [45% (v/v) ethanol, 10% (v/v) acetic acid, 0.25% (v/w) Coomassie Brilliant Blue R250] followed by destaining with a solution consisting of 30% (v/v) ethanol and 10% (v/v) acetic acid.

    IA:

    Article Title: Multiple layers of transcriptional regulation by PLZF in NKT-cell development
    Article Snippet: The following plasmids were used: pEF1a-BirA-V5his (a gift from K. Zhao, NIH, Bethesda), pCMV-HA-HDAC1 (a gift from J. Colgan, University of Iowa, Iowa City, IA), pcDNA3-Myc-CUL3 (a gift from Y. Xiong, University of North Carolina at Chapel Hill, Chapel Hill, NC), and pcDNA5-PLZF and pcDNA5-A/F/PLZF plasmids, constructed using standard molecular biology techniques. .. The following antibodies were used: anti-PLZF (AF2944; R & D Systems), anti-CUL3 (A301-109A; Bethyl Laboratories), anti-HDAC1 (ab-7028; Abcam), streptavidin-HRP (S2438; Sigma-Aldrich), anti–Flag-HRP (A8592; Sigma-Aldrich), anti-Bach2 (12809; CST), and anti-GAPDH (G9545; Sigma-Aldrich).

    Activated Clotting Time Assay:

    Article Title: Functional competence of a partially engaged GPCR–β-arrestin complex
    Article Snippet: ELISA based assembly of β2 V2 R–βarr1-Fab30 complex For ELISA based in-vitro assembly of β2 V2 R–βarr1–Fab/ScFv30 complexes, purified Fab/ScFv30 (in 20 mM Hepes, pH 7.4, 100 mM NaCl) was first immobilized on 96 well MaxiSorp polystyrene plates (Nunc) at room temperature for 1 h. Afterwards, potential non-specific binding sites in the wells were blocked by incubation with 1% BSA at room temperature for 1 h. Subsequently, mixture of ligand stimulated cell lysate (or purified receptor) was added to the wells and incubated at room temperature for 1 h. Wells were washed extensively using 20 mM Hepes, pH 7.4, 100 mM NaCl, 0.01% MNG and then incubated with 1:2,000 dilution of HRP-coupled anti-FLAG M2 antibody (Sigma, catalog number A8592). .. All the ELISA data are normalized with respect to the signal for Act β2 V2 Rphos complex which is treated as 100%.

    SDS Page:

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare). .. The purified rhPP2B dimer and rhPP2Cα were subjected to 13.5% and 12% SDS-PAGE, respectively, following which they were visualized using Coomassie Brilliant Blue R staining ( ).

    Article Title: Stimulator of interferon genes (STING) provides insect antiviral immunity by promoting Dredd caspase–mediated NF-κB activation
    Article Snippet: Sample from the co-immunoprecipitation assay and whole-cell lysate was separately resolved on 12% SDS-PAGE. .. After transfer to a polyvinylidene difluoride membrane (GE Healthcare), samples were immunoblotted with anti-HA-HRP antibody (catalogue no. A00169-40, Genscript, Nanjing, China), anti-FLAG-HRP antibody (catalogue no. A8592-2MG, Sigma), or GAPDH antibody (catalogue no. abs830030, Shanghai, China) following the standard procedure and then developed using SuperSignalTM West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) as described previously ( ).

    Software:

    Article Title: A simple vector system to improve performance and utilisation of recombinant antibodies
    Article Snippet: Where anti-FLAG and ECL detection was required, the membranes were incubated with 0.22 μg/ml of monoclonal Anti-FLAG® M2 peroxidase (HRP) (Sigma-Aldrich, Poole, UK) in M-PBS-T for 1 h at room temperature on an orbital shaker, and ECL western blotting substrate added according to the supplier's instructions (Pierce, Cramlington, UK). .. The membranes were scanned on the Typhoon scanner and analysed using ImageQuant software.

    Article Title: Cardiac L-type calcium channel (Cav1.2) associates with ? subunits
    Article Snippet: The anti-α2 antibody (D219) and the anti-Flag antibody (A8592) were from Sigma. .. Image quantification was performed using ImageQuant software (Amersham, Piscataway, NJ, USA).

    Electrophoresis:

    Article Title: Cardiac L-type calcium channel (Cav1.2) associates with ? subunits
    Article Snippet: The anti-α2 antibody (D219) and the anti-Flag antibody (A8592) were from Sigma. .. After electrophoresis, the proteins were transferred to nitrocellulose and probed with a primary antibody followed by an HRP-conjugated secondary antibody and ECL (Pierce, Rockford, IL, USA).

    Co-Immunoprecipitation Assay:

    Article Title: Stimulator of interferon genes (STING) provides insect antiviral immunity by promoting Dredd caspase–mediated NF-κB activation
    Article Snippet: Sample from the co-immunoprecipitation assay and whole-cell lysate was separately resolved on 12% SDS-PAGE. .. After transfer to a polyvinylidene difluoride membrane (GE Healthcare), samples were immunoblotted with anti-HA-HRP antibody (catalogue no. A00169-40, Genscript, Nanjing, China), anti-FLAG-HRP antibody (catalogue no. A8592-2MG, Sigma), or GAPDH antibody (catalogue no. abs830030, Shanghai, China) following the standard procedure and then developed using SuperSignalTM West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) as described previously ( ).

    Affinity Chromatography:

    Article Title: Targeting scFv-Fc-scTRAIL fusion proteins to tumor cells
    Article Snippet: Materials Receptor-Fc and scFv-Fc fusion proteins were produced with stably transfected HEK293T cells and purified from the supernatant by protein A affinity chromatography as described previously [ , ]. .. HRP- and PE-conjugated anti-FLAG antibodies were obtained from Sigma-Aldrich (A8592) and Miltenyi Biotec (130-101-576), respectively.

    In Vitro:

    Article Title: Functional competence of a partially engaged GPCR–β-arrestin complex
    Article Snippet: .. ELISA based assembly of β2 V2 R–βarr1-Fab30 complex For ELISA based in-vitro assembly of β2 V2 R–βarr1–Fab/ScFv30 complexes, purified Fab/ScFv30 (in 20 mM Hepes, pH 7.4, 100 mM NaCl) was first immobilized on 96 well MaxiSorp polystyrene plates (Nunc) at room temperature for 1 h. Afterwards, potential non-specific binding sites in the wells were blocked by incubation with 1% BSA at room temperature for 1 h. Subsequently, mixture of ligand stimulated cell lysate (or purified receptor) was added to the wells and incubated at room temperature for 1 h. Wells were washed extensively using 20 mM Hepes, pH 7.4, 100 mM NaCl, 0.01% MNG and then incubated with 1:2,000 dilution of HRP-coupled anti-FLAG M2 antibody (Sigma, catalog number A8592). .. After 1 h incubation, wells were extensively washed and assembly of the complex was visualized by adding 3,3′,5,5′-tetramethylbenzidine (TMB) ELISA (Genscript or Thermo).

    Article Title: Targeting scFv-Fc-scTRAIL fusion proteins to tumor cells
    Article Snippet: HRP- and PE-conjugated anti-FLAG antibodies were obtained from Sigma-Aldrich (A8592) and Miltenyi Biotec (130-101-576), respectively. .. For in vitro studies, bortezomib (BZB) was purchased from UBPBio (F1200).

    Produced:

    Article Title: Targeting scFv-Fc-scTRAIL fusion proteins to tumor cells
    Article Snippet: Materials Receptor-Fc and scFv-Fc fusion proteins were produced with stably transfected HEK293T cells and purified from the supernatant by protein A affinity chromatography as described previously [ , ]. .. HRP- and PE-conjugated anti-FLAG antibodies were obtained from Sigma-Aldrich (A8592) and Miltenyi Biotec (130-101-576), respectively.

    Immunoprecipitation:

    Article Title: Multiple layers of transcriptional regulation by PLZF in NKT-cell development
    Article Snippet: Paragraph title: Immunoprecipitation and Western Blot Analysis. ... The following antibodies were used: anti-PLZF (AF2944; R & D Systems), anti-CUL3 (A301-109A; Bethyl Laboratories), anti-HDAC1 (ab-7028; Abcam), streptavidin-HRP (S2438; Sigma-Aldrich), anti–Flag-HRP (A8592; Sigma-Aldrich), anti-Bach2 (12809; CST), and anti-GAPDH (G9545; Sigma-Aldrich).

    Article Title: Cardiac L-type calcium channel (Cav1.2) associates with ? subunits
    Article Snippet: Paragraph title: Immunoprecipitation and immunoblots ... The anti-α2 antibody (D219) and the anti-Flag antibody (A8592) were from Sigma.

    Recombinant:

    Article Title: A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation
    Article Snippet: The membrane was washed with TBST (three times, 25 ml) and incubated with α-FLAG-HRP (Sigma A-8592, diluted 1:3,000 in TBST) for 1 h. The membrane was washed with TBST (three times, 25 ml) and developed by using SuperSignal West Pico Chemiluminescent Substrate (Pierce). .. Incorporation of GalNAz into Recombinant GlyCAM-Ig.

    Article Title: Efficient production of recombinant PP2A at a low temperature using a baculovirus expression system
    Article Snippet: Infections were performed at total m.o.i. of 6 by premixing equal m.o.i.s of recombinant viruses, vHis-PP2B-Aα and vFLAG-PP2B-Bα. .. Expressed His8x -tagged hPP2B-Aα, FLAG-tagged hPP2B-Bα, and His8x -tagged hPP2Cα were detected by Western blot analysis using anti-polyhistidine-peroxidase conjugate (Sigma) and anti-FLAG M2 monoclonal antibody-peroxidase conjugate (Sigma) followed by the ECL system (GE Healthcare).

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    Millipore a8592
    A8592, supplied by Millipore, 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/a8592/product/Millipore
    Average 99 stars, based on 1 article reviews
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    a8592 - by Bioz Stars, 2020-04
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