streptavidin coated agarose beads  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 88

    Structured Review

    Millipore streptavidin coated agarose beads
    p120 regulates E-cadherin turnover at the cell membrane. (a) E-cadherin synthesis and processing in p120 knockdown cells. E-cadherin turnover was examined by pulse-chase analysis of parental and h p120 siRNA-expressing A431 cells. α- and β-Catenin processing from the same experiment are shown below. Chase times are indicated across the top. At chase time 0 (15 min after initiation of the pulse labeling), E-cadherin synthesis was identical in the presence and absence of p120 (a, compare E-cadherin bands). The processing of the pro- (pro-E-cad) and mature (E-cad.) forms were identical for at least 1 h. Soon thereafter, E-cadherin degradation was significantly accelerated in the absence of p120. (b) Analysis of total E-cadherin surface levels in parental and p120 knockdown A431 cells. The parental and p120 knockdown A431 cells were biotinylated for 20 min at 4°C to label surface cadherins. To specifically measure the surface levels, E-cadherin was first immunoprecipitated directly with E-cadherin mAb HECD-1. The sample was eluted with 0.5% SDS and then reprecipitated with <t>streptavidin-coated</t> beads to isolate the surface-labeled pool. E-cadherin levels at the surface in p120 knockdown cells (lane 1) are at least 100-fold diminished relative to the parental cells (lane 2). The result in lane 2 shows that the surface E-cadherin can be efficiently labeled (and detected) by this method. (c) Tracking the arrival of newly synthesized E-cadherin to the cell surface. The methods in a and b were combined to determine whether newly synthesized E-cadherin could transit to the cell surface in the absence of p120. The results in c were quantified by densitometry and represented graphically in d. Parental and p120 knockdown (h siRNA) cells were labeled with [ 35 S]methionine for 15 min, chased at 37°C for the times indicated across top, and placed on ice (4°C) to suspend trafficking. Cell surface proteins were immediately biotinylated at 4°C for 20 min as in b. Surface E-cadherin was then isolated as in b, and the nascent [ 35 S]methionine E-cadherin pool was visualized by SDS-PAGE and radiography. Nascent E-cadherin appeared at the surface at 30 min and peaked at 1 h. The absence of p120 had no effect on this result. Therefore, p120 is not required for E-cadherin synthesis or trafficking, but is essential to regulate E-cadherin turnover soon after its arrival at the cell surface.
    Streptavidin Coated Agarose Beads, 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/streptavidin coated agarose beads/product/Millipore
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    streptavidin coated agarose beads - by Bioz Stars, 2020-04
    88/100 stars

    Images

    1) Product Images from "A core function for p120-catenin in cadherin turnover"

    Article Title: A core function for p120-catenin in cadherin turnover

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.200307111

    p120 regulates E-cadherin turnover at the cell membrane. (a) E-cadherin synthesis and processing in p120 knockdown cells. E-cadherin turnover was examined by pulse-chase analysis of parental and h p120 siRNA-expressing A431 cells. α- and β-Catenin processing from the same experiment are shown below. Chase times are indicated across the top. At chase time 0 (15 min after initiation of the pulse labeling), E-cadherin synthesis was identical in the presence and absence of p120 (a, compare E-cadherin bands). The processing of the pro- (pro-E-cad) and mature (E-cad.) forms were identical for at least 1 h. Soon thereafter, E-cadherin degradation was significantly accelerated in the absence of p120. (b) Analysis of total E-cadherin surface levels in parental and p120 knockdown A431 cells. The parental and p120 knockdown A431 cells were biotinylated for 20 min at 4°C to label surface cadherins. To specifically measure the surface levels, E-cadherin was first immunoprecipitated directly with E-cadherin mAb HECD-1. The sample was eluted with 0.5% SDS and then reprecipitated with streptavidin-coated beads to isolate the surface-labeled pool. E-cadherin levels at the surface in p120 knockdown cells (lane 1) are at least 100-fold diminished relative to the parental cells (lane 2). The result in lane 2 shows that the surface E-cadherin can be efficiently labeled (and detected) by this method. (c) Tracking the arrival of newly synthesized E-cadherin to the cell surface. The methods in a and b were combined to determine whether newly synthesized E-cadherin could transit to the cell surface in the absence of p120. The results in c were quantified by densitometry and represented graphically in d. Parental and p120 knockdown (h siRNA) cells were labeled with [ 35 S]methionine for 15 min, chased at 37°C for the times indicated across top, and placed on ice (4°C) to suspend trafficking. Cell surface proteins were immediately biotinylated at 4°C for 20 min as in b. Surface E-cadherin was then isolated as in b, and the nascent [ 35 S]methionine E-cadherin pool was visualized by SDS-PAGE and radiography. Nascent E-cadherin appeared at the surface at 30 min and peaked at 1 h. The absence of p120 had no effect on this result. Therefore, p120 is not required for E-cadherin synthesis or trafficking, but is essential to regulate E-cadherin turnover soon after its arrival at the cell surface.
    Figure Legend Snippet: p120 regulates E-cadherin turnover at the cell membrane. (a) E-cadherin synthesis and processing in p120 knockdown cells. E-cadherin turnover was examined by pulse-chase analysis of parental and h p120 siRNA-expressing A431 cells. α- and β-Catenin processing from the same experiment are shown below. Chase times are indicated across the top. At chase time 0 (15 min after initiation of the pulse labeling), E-cadherin synthesis was identical in the presence and absence of p120 (a, compare E-cadherin bands). The processing of the pro- (pro-E-cad) and mature (E-cad.) forms were identical for at least 1 h. Soon thereafter, E-cadherin degradation was significantly accelerated in the absence of p120. (b) Analysis of total E-cadherin surface levels in parental and p120 knockdown A431 cells. The parental and p120 knockdown A431 cells were biotinylated for 20 min at 4°C to label surface cadherins. To specifically measure the surface levels, E-cadherin was first immunoprecipitated directly with E-cadherin mAb HECD-1. The sample was eluted with 0.5% SDS and then reprecipitated with streptavidin-coated beads to isolate the surface-labeled pool. E-cadherin levels at the surface in p120 knockdown cells (lane 1) are at least 100-fold diminished relative to the parental cells (lane 2). The result in lane 2 shows that the surface E-cadherin can be efficiently labeled (and detected) by this method. (c) Tracking the arrival of newly synthesized E-cadherin to the cell surface. The methods in a and b were combined to determine whether newly synthesized E-cadherin could transit to the cell surface in the absence of p120. The results in c were quantified by densitometry and represented graphically in d. Parental and p120 knockdown (h siRNA) cells were labeled with [ 35 S]methionine for 15 min, chased at 37°C for the times indicated across top, and placed on ice (4°C) to suspend trafficking. Cell surface proteins were immediately biotinylated at 4°C for 20 min as in b. Surface E-cadherin was then isolated as in b, and the nascent [ 35 S]methionine E-cadherin pool was visualized by SDS-PAGE and radiography. Nascent E-cadherin appeared at the surface at 30 min and peaked at 1 h. The absence of p120 had no effect on this result. Therefore, p120 is not required for E-cadherin synthesis or trafficking, but is essential to regulate E-cadherin turnover soon after its arrival at the cell surface.

    Techniques Used: Pulse Chase, Expressing, Labeling, Immunoprecipitation, Synthesized, Isolation, SDS Page

    Related Articles

    Clone Assay:

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: Thus, we cloned a 778 base pair upstream sequence of DNA from the translational initiation site of the msdps2 gene, in mc2 155 genomic DNA sequence, assuming that the promoter element will be a part of the same. .. The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich).

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. RGC-32 proteins were obtained by expression of pETDuet-1-RGC32 in E. coli and purified by His-Spin Protein Miniprep kit (Zymo Research). pETDuet-1-RGC32 was cloned by insertion of full-length mouse RGC-32 cDNA into pETDuet-1 expression vector at BamHI/SalI sites.

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: To produce PIF4-his protein, full-length PIF4 coding sequence was amplified and cloned into pET28a vector. .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    Centrifugation:

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C. .. The DNA-beads were incubated with purified protein for 1 h and then collected by centrifugation and washed three times with DNA binding buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 5 mM EDTA, 0.05% Nonidet P-40).

    Amplification:

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: To produce PIF4-his protein, full-length PIF4 coding sequence was amplified and cloned into pET28a vector. .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    Autoradiography:

    Article Title: A core function for p120-catenin in cadherin turnover
    Article Snippet: At the end of the chase, cell surface proteins were labeled with 1 mg/ml EZ-Link sulfo-NHS-SS-biotin (Pierce Chemical Co.) at 4°C for 30 min. E-cadherin was immunoprecipitated from NP-40 cell lysates, and surface cadherin was detected by dividing the E-cadherin immunoprecipitations in half, eluting E-cadherin from the beads with 0.5% SDS, reconstituting elutions in 50 mM Tris-HCl (pH 7.5), and pulling down biotinylated E-cadherin with 10 μl per sample of packed streptavidin-coated agarose beads (Sigma-Aldrich) for 1 h at 4°C. .. Samples were washed three times with 50 mM Tris-HCl (pH 7.5), and protein was eluted with 2× Laemmli sample buffer and analyzed by SDS-PAGE and autoradiography as described previously ( ).

    Construct:

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich). .. In order to quantitate the differential affinity of the two sigma factors reconstituted holo RNA polymerases namely EσA and EσB for msdps2 promoter, a calibration curve was constructed ( ). shows the comparative affinity for EσA and EσB towards msdps2 promoter.

    Incubation:

    Article Title: Pathway specific modulation of S1P1 receptor signalling in rat and human astrocytes
    Article Snippet: Cultured human astrocytes were serum starved for 4 h, pretreated with or without 100 μg·mL−1 MNP301 for 1 h and then incubated with or without 1 μM pFTY720 for 1 h. Astrocytes were then incubated with 0.5 mg·mL−1 biotin for 30 min at 4°C, after which the cells were scraped in PTxE buffer (PBS, 1% Triton X-100, 0.1 mM EDTA, pH 7.4) at 4°C. .. Sample biotin was immunoprecipitated for 2 h at 4°C using streptavidin-coated agarose beads (Sigma), then analysed by Western blotting.

    Article Title: Role of Per1 and the mineralocorticoid receptor in the coordinate regulation of ?ENaC in renal cortical collecting duct cells
    Article Snippet: .. For DNA affinity purification assay (DAPA), Probes were immobilized on 50 μ l of streptavidin-coated agarose beads (Sigma) and incubated with 175 μ g of nuclear mpkCCDc14 extracts either treated with vehicle (ethanol) or 1 μ M aldosterone for 24 h in the presence of freshly prepared protease inhibitors (Fischer) for 2 h at 4°C with end-over-end rotation. ..

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: .. Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. The agarose beads were pelleted and washed with cold 1× PBS three times, and proteins bound to the beads were eluted and immunoblotted with Smad3, Smad4, or RGC-32 antibodies.

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C. .. The DNA-beads were incubated with purified protein for 1 h and then collected by centrifugation and washed three times with DNA binding buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 5 mM EDTA, 0.05% Nonidet P-40).

    Article Title: Mycobacterial RNA Polymerase Requires a U-Tract at Intrinsic Terminators and Is Aided by NusG at Suboptimal Terminators
    Article Snippet: .. A linear DNA template (10 nM) containing biotin at the 5′ end of the nontemplate strand and 25 nM Mbo RNAP holoenzyme were combined with 10 µl of streptavidin-coated agarose beads (Sigma) and incubated for 15 min at 37°C in RB (30 mM Tris-HCl [pH 7.5], 80 mM NaCl, 1 mM DTT, 10 mM MgCl2 , 0.1 mM EDTA, 0.025% Triton X-100, 2% glycerol, 0.05 mg BSA/ml). .. Eco RIQ111 (500 nM) was added to complexes and incubated for 10 min at 37°C.

    In Silico:

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich). .. The western blots were scanned using Multi Gauge V2.3 software (in silico ) and the transcript band intensities were quantitated through densitometry.

    Expressing:

    Article Title: UV-C irradiation disrupts platelet surface disulfide bonds and activates the platelet integrin ?IIb?3
    Article Snippet: A control experiment with platelets from a Glanzmann patient lacking expression of αIIbβ3 proved the specificity of the β3 antibody, since the immunoreactive band (running above 95 kDa under reduced conditions) was absent in this sample. .. The adenylate cyclase stimulator forskolin; the PKC inhibitors Ro 31-8220, Rottlerin, and staurosporin; the PI3-kinase inhibitor wortmannin; and streptavidin-coated agarose beads were obtained from Sigma (Zwijndrecht, The Netherlands).

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. RGC-32 proteins were obtained by expression of pETDuet-1-RGC32 in E. coli and purified by His-Spin Protein Miniprep kit (Zymo Research). pETDuet-1-RGC32 was cloned by insertion of full-length mouse RGC-32 cDNA into pETDuet-1 expression vector at BamHI/SalI sites.

    Western Blot:

    Article Title: Pathway specific modulation of S1P1 receptor signalling in rat and human astrocytes
    Article Snippet: .. Sample biotin was immunoprecipitated for 2 h at 4°C using streptavidin-coated agarose beads (Sigma), then analysed by Western blotting. .. Astrocytes were trypsinized, diluted and seeded into 24-well cell culture plates (MidSci, Saint Louis, MO, USA) at a cell density of 5 × 105 cells·mL−1 .

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich). .. The western blots were scanned using Multi Gauge V2.3 software (in silico ) and the transcript band intensities were quantitated through densitometry.

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. The cDNA insertion was verified by sequencing and the purity of RGC-32 protein was verified by Coomassie Blue staining and Western blot using RGC-32 antibody.

    Article Title: A naturally occurring truncated Cav1.2 α1-subunit inhibits Ca2+ current in A7r5 cells
    Article Snippet: Biotinylated proteins were recovered from the cell lysates with streptavidin-coated agarose beads (Sigma Chemical, Rockford, IL). .. Biotinylated and nonbiotinylated proteins plus precleared whole cell lysates were prepared for Western blotting, as described above.

    Pulse Chase:

    Article Title: A core function for p120-catenin in cadherin turnover
    Article Snippet: Paragraph title: Pulse chase, biotinylation, and cell surface trafficking ... At the end of the chase, cell surface proteins were labeled with 1 mg/ml EZ-Link sulfo-NHS-SS-biotin (Pierce Chemical Co.) at 4°C for 30 min. E-cadherin was immunoprecipitated from NP-40 cell lysates, and surface cadherin was detected by dividing the E-cadherin immunoprecipitations in half, eluting E-cadherin from the beads with 0.5% SDS, reconstituting elutions in 50 mM Tris-HCl (pH 7.5), and pulling down biotinylated E-cadherin with 10 μl per sample of packed streptavidin-coated agarose beads (Sigma-Aldrich) for 1 h at 4°C.

    Concentration Assay:

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich). .. We reconstituted M. smegmatis core RNA polymerase with different concentration of σA or σB , named as EσA and EσB respectively and carried out pull-down experiments and probed with antibodies against the respective sigma factors.

    Cell Culture:

    Article Title: Pathway specific modulation of S1P1 receptor signalling in rat and human astrocytes
    Article Snippet: Cultured human astrocytes were serum starved for 4 h, pretreated with or without 100 μg·mL−1 MNP301 for 1 h and then incubated with or without 1 μM pFTY720 for 1 h. Astrocytes were then incubated with 0.5 mg·mL−1 biotin for 30 min at 4°C, after which the cells were scraped in PTxE buffer (PBS, 1% Triton X-100, 0.1 mM EDTA, pH 7.4) at 4°C. .. Sample biotin was immunoprecipitated for 2 h at 4°C using streptavidin-coated agarose beads (Sigma), then analysed by Western blotting.

    Polymerase Chain Reaction:

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C. .. The DNA-beads were incubated with purified protein for 1 h and then collected by centrifugation and washed three times with DNA binding buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 5 mM EDTA, 0.05% Nonidet P-40).

    Sonication:

    Article Title: Pathway specific modulation of S1P1 receptor signalling in rat and human astrocytes
    Article Snippet: The cell suspension was then sonicated for 10 × 2 s (Astrason). .. Sample biotin was immunoprecipitated for 2 h at 4°C using streptavidin-coated agarose beads (Sigma), then analysed by Western blotting.

    Affinity Purification:

    Article Title: Role of Per1 and the mineralocorticoid receptor in the coordinate regulation of ?ENaC in renal cortical collecting duct cells
    Article Snippet: .. For DNA affinity purification assay (DAPA), Probes were immobilized on 50 μ l of streptavidin-coated agarose beads (Sigma) and incubated with 175 μ g of nuclear mpkCCDc14 extracts either treated with vehicle (ethanol) or 1 μ M aldosterone for 24 h in the presence of freshly prepared protease inhibitors (Fischer) for 2 h at 4°C with end-over-end rotation. ..

    Binding Assay:

    Article Title: UV-C irradiation disrupts platelet surface disulfide bonds and activates the platelet integrin ?IIb?3
    Article Snippet: The monoclonal antibody PAC-1 binding to activated αIIbβ3 (conjugated to fluorescein isothiocyanate [FITC]) and the anti-β3 antibody (clone 1) used for immunoblotting were purchased from BD Biosciences (San Jose, CA). .. The adenylate cyclase stimulator forskolin; the PKC inhibitors Ro 31-8220, Rottlerin, and staurosporin; the PI3-kinase inhibitor wortmannin; and streptavidin-coated agarose beads were obtained from Sigma (Zwijndrecht, The Netherlands).

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: .. Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. The agarose beads were pelleted and washed with cold 1× PBS three times, and proteins bound to the beads were eluted and immunoblotted with Smad3, Smad4, or RGC-32 antibodies.

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C. .. The DNA-beads were incubated with purified protein for 1 h and then collected by centrifugation and washed three times with DNA binding buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 5 mM EDTA, 0.05% Nonidet P-40).

    Article Title: MOLECULAR CHARACTERIZATION OF HTLV-1 TAX INTERACTION WITH THE KIX DOMAIN OF CBP/p300
    Article Snippet: DNA pull-down experiments were performed using streptavidin-coated agarose beads (Novagen). .. The amount of DNA bound was quantified by measuring the A260 of the DNA-containing supernatant before and after streptavidin-agarose bead binding.

    In Vivo:

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: However, it was difficult to characterize the same with in vivo primer extension method , as we do not know under what condition the transcription of msdps2 is activated. .. The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich).

    Pull Down Assay:

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: Paragraph title: In Vitro DNA:Protein Pull-Down Assay ... Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    Isolation:

    Article Title: Role of Per1 and the mineralocorticoid receptor in the coordinate regulation of ?ENaC in renal cortical collecting duct cells
    Article Snippet: Nuclear extracts, DNA affinity purification assays (DAPA), and immunoblotting Nuclear and cytosolic extracts were isolated using the NE-PER kit (Pierce) according to the manufacturer's instructions. .. For DNA affinity purification assay (DAPA), Probes were immobilized on 50 μ l of streptavidin-coated agarose beads (Sigma) and incubated with 175 μ g of nuclear mpkCCDc14 extracts either treated with vehicle (ethanol) or 1 μ M aldosterone for 24 h in the presence of freshly prepared protease inhibitors (Fischer) for 2 h at 4°C with end-over-end rotation.

    Article Title: A naturally occurring truncated Cav1.2 α1-subunit inhibits Ca2+ current in A7r5 cells
    Article Snippet: After the cells were washed and quenched, whole cell protein lysates were isolated using Triton lysis buffer (150 mmol/l NaCl, 1.5 mmol/l MgCl2 , 20 mmol/l HEPES, 1% Triton X-100, and 10% glycerol, pH 7.5) with protease and phosphatase inhibitors. .. Biotinylated proteins were recovered from the cell lysates with streptavidin-coated agarose beads (Sigma Chemical, Rockford, IL).

    Labeling:

    Article Title: A core function for p120-catenin in cadherin turnover
    Article Snippet: .. At the end of the chase, cell surface proteins were labeled with 1 mg/ml EZ-Link sulfo-NHS-SS-biotin (Pierce Chemical Co.) at 4°C for 30 min. E-cadherin was immunoprecipitated from NP-40 cell lysates, and surface cadherin was detected by dividing the E-cadherin immunoprecipitations in half, eluting E-cadherin from the beads with 0.5% SDS, reconstituting elutions in 50 mM Tris-HCl (pH 7.5), and pulling down biotinylated E-cadherin with 10 μl per sample of packed streptavidin-coated agarose beads (Sigma-Aldrich) for 1 h at 4°C. .. Samples were washed three times with 50 mM Tris-HCl (pH 7.5), and protein was eluted with 2× Laemmli sample buffer and analyzed by SDS-PAGE and autoradiography as described previously ( ).

    Article Title: UV-C irradiation disrupts platelet surface disulfide bonds and activates the platelet integrin ?IIb?3
    Article Snippet: The adenylate cyclase stimulator forskolin; the PKC inhibitors Ro 31-8220, Rottlerin, and staurosporin; the PI3-kinase inhibitor wortmannin; and streptavidin-coated agarose beads were obtained from Sigma (Zwijndrecht, The Netherlands). .. Monoclonal antibody directed against CD61 (β3) or isotype-matched control IgG1, both labeled with FITC, were purchased from Sanquin (Amsterdam, The Netherlands).

    Article Title: Involvement of KSRP in the post-transcriptional regulation of human iNOS expression-complex interplay of KSRP with TTP and HuR
    Article Snippet: Materials Trypsin-, glutamine-, and pyruvate-solutions, phenylmethylsulfonyl fluoride, leupeptin, aprotinin, agarose, isopropyl-β-d -thiogalactopyranoside, tRNA, BSA, actinomycin D, streptavidin-coated agarose beads, horseradish-peroxidase-coupled anti-goat and anti-mouse IgG were purchased from Sigma, Deisenhofen, Germany. .. RNase A, RNase T1, DNase I, T3 and T7 RNA polymerase and the biotin RNA labeling mix were obtained from Roche Diagnostics, Mannheim, Germany.

    Purification:

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. RGC-32 proteins were obtained by expression of pETDuet-1-RGC32 in E. coli and purified by His-Spin Protein Miniprep kit (Zymo Research). pETDuet-1-RGC32 was cloned by insertion of full-length mouse RGC-32 cDNA into pETDuet-1 expression vector at BamHI/SalI sites.

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: The recombinant PIF4-his protein was purified using NI-NTA resin according to the manufacturer’s instructions (Qiagen, USA). .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    Sequencing:

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: We have shown before that both M. tuberculosis σA and σB share significant sequence homology with that of M. smegmatis . .. The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich).

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. The cDNA insertion was verified by sequencing and the purity of RGC-32 protein was verified by Coomassie Blue staining and Western blot using RGC-32 antibody.

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: To produce PIF4-his protein, full-length PIF4 coding sequence was amplified and cloned into pET28a vector. .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    Staining:

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. The cDNA insertion was verified by sequencing and the purity of RGC-32 protein was verified by Coomassie Blue staining and Western blot using RGC-32 antibody.

    SDS Page:

    Article Title: A core function for p120-catenin in cadherin turnover
    Article Snippet: At the end of the chase, cell surface proteins were labeled with 1 mg/ml EZ-Link sulfo-NHS-SS-biotin (Pierce Chemical Co.) at 4°C for 30 min. E-cadherin was immunoprecipitated from NP-40 cell lysates, and surface cadherin was detected by dividing the E-cadherin immunoprecipitations in half, eluting E-cadherin from the beads with 0.5% SDS, reconstituting elutions in 50 mM Tris-HCl (pH 7.5), and pulling down biotinylated E-cadherin with 10 μl per sample of packed streptavidin-coated agarose beads (Sigma-Aldrich) for 1 h at 4°C. .. Samples were washed three times with 50 mM Tris-HCl (pH 7.5), and protein was eluted with 2× Laemmli sample buffer and analyzed by SDS-PAGE and autoradiography as described previously ( ).

    Plasmid Preparation:

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight. .. RGC-32 proteins were obtained by expression of pETDuet-1-RGC32 in E. coli and purified by His-Spin Protein Miniprep kit (Zymo Research). pETDuet-1-RGC32 was cloned by insertion of full-length mouse RGC-32 cDNA into pETDuet-1 expression vector at BamHI/SalI sites.

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: To produce PIF4-his protein, full-length PIF4 coding sequence was amplified and cloned into pET28a vector. .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    Software:

    Article Title: A core function for p120-catenin in cadherin turnover
    Article Snippet: At the end of the chase, cell surface proteins were labeled with 1 mg/ml EZ-Link sulfo-NHS-SS-biotin (Pierce Chemical Co.) at 4°C for 30 min. E-cadherin was immunoprecipitated from NP-40 cell lysates, and surface cadherin was detected by dividing the E-cadherin immunoprecipitations in half, eluting E-cadherin from the beads with 0.5% SDS, reconstituting elutions in 50 mM Tris-HCl (pH 7.5), and pulling down biotinylated E-cadherin with 10 μl per sample of packed streptavidin-coated agarose beads (Sigma-Aldrich) for 1 h at 4°C. .. Quantification was performed by densitometry using Image Gage software (Fujifilm Inc.).

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich). .. The western blots were scanned using Multi Gauge V2.3 software (in silico ) and the transcript band intensities were quantitated through densitometry.

    Recombinant:

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: The recombinant PIF4-his protein was purified using NI-NTA resin according to the manufacturer’s instructions (Qiagen, USA). .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    In Vitro:

    Article Title: The Mycobacterial MsDps2 Protein Is a Nucleoid-Forming DNA Binding Protein Regulated by Sigma Factors ?A and ?B
    Article Snippet: Preliminary multiple round in vitro transcription on this template showed appreciable RNA product with the core RNA polymerase of M. smegmatis reconstituted with M. tuberculosis σA and σB factors (not shown). .. The DNA fragment was then immobilized on streptavidin coated agarose beads (Sigma Aldrich).

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: Paragraph title: In Vitro DNA:Protein Pull-Down Assay ... Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C.

    Protein Binding:

    Article Title: Response Gene to Complement 32 Is Essential for Fibroblast Activation in Renal Fibrosis *
    Article Snippet: DNA protein binding assay was performed as described ( , ). .. Briefly, 5 μg of biotin-labeled double-strand oligonucleotides, and 500 ng of Smad3, Smad4 (SignalChem), and/or RGC-32 proteins were incubated in a binding buffer (0.1% Triton X-100, 4% glycerol, 1 m m EDTA, 10 m m DTT, 10 m m Tris) at 37 °C for 1 h. Streptavidin-coated agarose beads (Sigma) were then added followed by end-over-end rotation at 4 °C overnight.

    Produced:

    Article Title: Interactions between HLH and bHLH Factors Modulate Light-Regulated Plant Development
    Article Snippet: .. Biotinylated PIL1 promoter DNA was produced by PCR using biotinylated oligos, bound to the streptavidin-coated agarose beads (Sigma, USA) for at least 1 h in advance at 4°C. .. The DNA-beads were incubated with purified protein for 1 h and then collected by centrifugation and washed three times with DNA binding buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 5 mM EDTA, 0.05% Nonidet P-40).

    Immunoprecipitation:

    Article Title: Pathway specific modulation of S1P1 receptor signalling in rat and human astrocytes
    Article Snippet: .. Sample biotin was immunoprecipitated for 2 h at 4°C using streptavidin-coated agarose beads (Sigma), then analysed by Western blotting. .. Astrocytes were trypsinized, diluted and seeded into 24-well cell culture plates (MidSci, Saint Louis, MO, USA) at a cell density of 5 × 105 cells·mL−1 .

    Article Title: A core function for p120-catenin in cadherin turnover
    Article Snippet: .. At the end of the chase, cell surface proteins were labeled with 1 mg/ml EZ-Link sulfo-NHS-SS-biotin (Pierce Chemical Co.) at 4°C for 30 min. E-cadherin was immunoprecipitated from NP-40 cell lysates, and surface cadherin was detected by dividing the E-cadherin immunoprecipitations in half, eluting E-cadherin from the beads with 0.5% SDS, reconstituting elutions in 50 mM Tris-HCl (pH 7.5), and pulling down biotinylated E-cadherin with 10 μl per sample of packed streptavidin-coated agarose beads (Sigma-Aldrich) for 1 h at 4°C. .. Samples were washed three times with 50 mM Tris-HCl (pH 7.5), and protein was eluted with 2× Laemmli sample buffer and analyzed by SDS-PAGE and autoradiography as described previously ( ).

    Lysis:

    Article Title: A naturally occurring truncated Cav1.2 α1-subunit inhibits Ca2+ current in A7r5 cells
    Article Snippet: After the cells were washed and quenched, whole cell protein lysates were isolated using Triton lysis buffer (150 mmol/l NaCl, 1.5 mmol/l MgCl2 , 20 mmol/l HEPES, 1% Triton X-100, and 10% glycerol, pH 7.5) with protease and phosphatase inhibitors. .. Biotinylated proteins were recovered from the cell lysates with streptavidin-coated agarose beads (Sigma Chemical, Rockford, IL).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Millipore streptavidin agarose beads
    Proline-rich reading array screen and peptide pulldown. (A) Use of biotinylated eVP40 WT (MRRVILPTAPPEYMEAI[Lys-biotin]) peptide (50 μg) to screen a proline-rich reading array. The GST-WW domain fusion proteins are arrayed in duplicate and at different angles, as indicated in enlarged box C. Box C shows duplicate samples of all four WW domains from WWP1, WWP2, and ITCH as indicated. Additional positive interactions are indicated in the highlighted red boxes and ovals (A to H). The eVP40 mutant peptide (MRRVILPTAAAEAMEAI[Lys-biotin]) did not interact with any GST-WW domain fusion protein (data not shown). (B) Exogenously expressed FLAG-tagged WWP1-WT was pulled down with <t>streptavidin</t> beads bound to either eVP40 WT (WT) or PPXY mutant (mut) peptides and detected by Western blotting using anti-Flag antiserum (top). Expression controls for WWP1 and actin are shown (bottom).
    Streptavidin Agarose Beads, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 293 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin agarose beads/product/Millipore
    Average 97 stars, based on 293 article reviews
    Price from $9.99 to $1999.99
    streptavidin agarose beads - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    Image Search Results


    Proline-rich reading array screen and peptide pulldown. (A) Use of biotinylated eVP40 WT (MRRVILPTAPPEYMEAI[Lys-biotin]) peptide (50 μg) to screen a proline-rich reading array. The GST-WW domain fusion proteins are arrayed in duplicate and at different angles, as indicated in enlarged box C. Box C shows duplicate samples of all four WW domains from WWP1, WWP2, and ITCH as indicated. Additional positive interactions are indicated in the highlighted red boxes and ovals (A to H). The eVP40 mutant peptide (MRRVILPTAAAEAMEAI[Lys-biotin]) did not interact with any GST-WW domain fusion protein (data not shown). (B) Exogenously expressed FLAG-tagged WWP1-WT was pulled down with streptavidin beads bound to either eVP40 WT (WT) or PPXY mutant (mut) peptides and detected by Western blotting using anti-Flag antiserum (top). Expression controls for WWP1 and actin are shown (bottom).

    Journal: Journal of Virology

    Article Title: Ubiquitin Ligase WWP1 Interacts with Ebola Virus VP40 To Regulate Egress

    doi: 10.1128/JVI.00812-17

    Figure Lengend Snippet: Proline-rich reading array screen and peptide pulldown. (A) Use of biotinylated eVP40 WT (MRRVILPTAPPEYMEAI[Lys-biotin]) peptide (50 μg) to screen a proline-rich reading array. The GST-WW domain fusion proteins are arrayed in duplicate and at different angles, as indicated in enlarged box C. Box C shows duplicate samples of all four WW domains from WWP1, WWP2, and ITCH as indicated. Additional positive interactions are indicated in the highlighted red boxes and ovals (A to H). The eVP40 mutant peptide (MRRVILPTAAAEAMEAI[Lys-biotin]) did not interact with any GST-WW domain fusion protein (data not shown). (B) Exogenously expressed FLAG-tagged WWP1-WT was pulled down with streptavidin beads bound to either eVP40 WT (WT) or PPXY mutant (mut) peptides and detected by Western blotting using anti-Flag antiserum (top). Expression controls for WWP1 and actin are shown (bottom).

    Article Snippet: Streptavidin agarose beads (Millipore) were prewashed once with 1× mild buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% NP-40, 5 mM EDTA, 5 mM EGTA, 15 mM MgCl2 ), and 15 μg of the WT or PPXY mutant eVP40 peptide was incubated with the prewashed streptavidin beads in 500 μl of 1× mild buffer for 1 h at 4°C with rocking.

    Techniques: Mutagenesis, Western Blot, Expressing

    The rate of cell surface expression/appearance/transport of BRI2 is reduced in the absence of N-glycosylation. Wild-type mycBRI2 or mycBRI2/N170A was expressed in HEK293 cells. The newly synthesized proteins were labeled with 35 S in radiolabeling medium for 2 h (pulse) at 16°C and then were incubated in non-radiolabeling medium for 0′, 20′, 40′ and 60′ (chase). ( A ) Cell surface proteins were labeled with biotin and precipitated with streptavidin beads. Precipitated cell surface proteins were eluted from the beads and immunoprecipitated with 9B11 antibody against the myc epitope before electrophoresis and autoradiography. ( B ) Immunoprecipitation of cell extracts with 9B11, electrophoresis and autoradiography were performed to verify the expression levels of BRI2.

    Journal: Glycobiology

    Article Title: Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10

    doi: 10.1093/glycob/cwr097

    Figure Lengend Snippet: The rate of cell surface expression/appearance/transport of BRI2 is reduced in the absence of N-glycosylation. Wild-type mycBRI2 or mycBRI2/N170A was expressed in HEK293 cells. The newly synthesized proteins were labeled with 35 S in radiolabeling medium for 2 h (pulse) at 16°C and then were incubated in non-radiolabeling medium for 0′, 20′, 40′ and 60′ (chase). ( A ) Cell surface proteins were labeled with biotin and precipitated with streptavidin beads. Precipitated cell surface proteins were eluted from the beads and immunoprecipitated with 9B11 antibody against the myc epitope before electrophoresis and autoradiography. ( B ) Immunoprecipitation of cell extracts with 9B11, electrophoresis and autoradiography were performed to verify the expression levels of BRI2.

    Article Snippet: The cell extracts were centrifuged at 15,000 × g for 30 min and supernatants were incubated with 50 μL of streptavidin–agarose beads (Millipore) for 1 h at 4°C.

    Techniques: Expressing, Synthesized, Labeling, Radioactivity, Incubation, Immunoprecipitation, Electrophoresis, Autoradiography

    Inhibition of N-glycosylation of BRI2 inhibits its expression at the cell surface. Wild-type mycBRI2 or mycBRI2/N170A was expressed in HEK293 cells. Cell surface proteins were labeled with biotin (lanes 1 and 2) or were not labeled (lanes 3 and 4), as a control for biotinylation specificity. ( A ) Cell extracts were precipitated with streptavidin beads and analyzed with western blot against myc with 9B11 antibody. ( B ) Cell extracts were directly analyzed with western blot as a control for protein expression. The two immunoreactive bands of BRI2 proteins correspond to the furin-cleaved and the non-cleaved wild-type mycBRI2 or mycBRI2/N170A.

    Journal: Glycobiology

    Article Title: Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10

    doi: 10.1093/glycob/cwr097

    Figure Lengend Snippet: Inhibition of N-glycosylation of BRI2 inhibits its expression at the cell surface. Wild-type mycBRI2 or mycBRI2/N170A was expressed in HEK293 cells. Cell surface proteins were labeled with biotin (lanes 1 and 2) or were not labeled (lanes 3 and 4), as a control for biotinylation specificity. ( A ) Cell extracts were precipitated with streptavidin beads and analyzed with western blot against myc with 9B11 antibody. ( B ) Cell extracts were directly analyzed with western blot as a control for protein expression. The two immunoreactive bands of BRI2 proteins correspond to the furin-cleaved and the non-cleaved wild-type mycBRI2 or mycBRI2/N170A.

    Article Snippet: The cell extracts were centrifuged at 15,000 × g for 30 min and supernatants were incubated with 50 μL of streptavidin–agarose beads (Millipore) for 1 h at 4°C.

    Techniques: Inhibition, Expressing, Labeling, Western Blot

    A19 phosphorylation. (A) Western blot analysis. BS-C-1 cells were infected with vFS-A11 and vFS-A19 in the presence of 100 μCi/ml 32 P i . After 18 h, the cells were lysed, and the soluble extract was bound to streptavidin-agarose beads. Bound proteins

    Journal: Journal of Virology

    Article Title: Vaccinia Virus A19 Protein Participates in the Transformation of Spherical Immature Particles to Barrel-Shaped Infectious Virions

    doi: 10.1128/JVI.01258-13

    Figure Lengend Snippet: A19 phosphorylation. (A) Western blot analysis. BS-C-1 cells were infected with vFS-A11 and vFS-A19 in the presence of 100 μCi/ml 32 P i . After 18 h, the cells were lysed, and the soluble extract was bound to streptavidin-agarose beads. Bound proteins

    Article Snippet: Soluble extracts obtained by low-speed centrifugation were allowed to bind to streptavidin-agarose beads (Millipore, Billerica, MA) for 3 h at 4°C.

    Techniques: Western Blot, Infection

    Expression and characterization of the A19 protein. (A) Schematic representation of the DNA construct used for generating recombinant vFS-A19. The FLAG- and streptavidin-binding peptide tag fused at the N terminus of the A19 ORF (FS) is indicated. The

    Journal: Journal of Virology

    Article Title: Vaccinia Virus A19 Protein Participates in the Transformation of Spherical Immature Particles to Barrel-Shaped Infectious Virions

    doi: 10.1128/JVI.01258-13

    Figure Lengend Snippet: Expression and characterization of the A19 protein. (A) Schematic representation of the DNA construct used for generating recombinant vFS-A19. The FLAG- and streptavidin-binding peptide tag fused at the N terminus of the A19 ORF (FS) is indicated. The

    Article Snippet: Soluble extracts obtained by low-speed centrifugation were allowed to bind to streptavidin-agarose beads (Millipore, Billerica, MA) for 3 h at 4°C.

    Techniques: Expressing, Construct, Recombinant, Binding Assay

    Analysis of viral PPxY-host WW-domain interactions between BAG3 and LFV-Z by peptide pull-down assays. ( A ) Flow chart of the peptide pull-down assay using LFV-Z peptides and cell lysates expressing BAG3-WT; ( B ) Schematic diagram of BAG3-WT, BAG3-ΔN, and BAG3-ΔC mutants with the various domains highlighted in color and amino acid positions indicated; ( C ) Western blot of peptide pull-down assay using streptavidin agarose beads conjugated with either the LFV-Z WT or LFV-Z PY mutant peptide. BAG3 proteins were detected using anti-c-myc antibody (top blot). Expression controls for BAG3 and actin are shown in the bottom blot. These results are from 1 of 2 independent experiments.

    Journal: Diseases

    Article Title: Host Protein BAG3 is a Negative Regulator of Lassa VLP Egress

    doi: 10.3390/diseases6030064

    Figure Lengend Snippet: Analysis of viral PPxY-host WW-domain interactions between BAG3 and LFV-Z by peptide pull-down assays. ( A ) Flow chart of the peptide pull-down assay using LFV-Z peptides and cell lysates expressing BAG3-WT; ( B ) Schematic diagram of BAG3-WT, BAG3-ΔN, and BAG3-ΔC mutants with the various domains highlighted in color and amino acid positions indicated; ( C ) Western blot of peptide pull-down assay using streptavidin agarose beads conjugated with either the LFV-Z WT or LFV-Z PY mutant peptide. BAG3 proteins were detected using anti-c-myc antibody (top blot). Expression controls for BAG3 and actin are shown in the bottom blot. These results are from 1 of 2 independent experiments.

    Article Snippet: Briefly, extracts from HEK293T cells expressing either BAG3-WT, BAG3-ΔN, or BAG3-ΔC ( B) were incubated with streptavidin agarose beads bound with either the LFV-Z-WT or LFV-Z-mutant peptides.

    Techniques: Flow Cytometry, Pull Down Assay, Expressing, Western Blot, Mutagenesis