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Boehringer Mannheim nonidet p 40
Nonidet P 40, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Labeling:

Article Title: Human Cytomegalovirus Inhibits Transcription of the CC Chemokine MCP-1 Gene
Article Snippet: Medium was collected from infected HFF cultures, and the supernatant was assayed by enzyme-linked immunosorbent assay (ELISA), using a Quantikine plate specific for human MCP-1 (R & D Systems) in accordance with the manufacturer’s protocol. .. For immunoprecipitations, confluent 10-cm-diameter dishes of HFFs were labeled with [35 S]methionine for 1 h and lysed in 0.5 ml of lysis buffer (50 mM Tris-Cl [pH 8.0], 150 mM NaCl, 0.1% sodium dodecyl sulfate, 1.0% Nonidet P-40, 0.5% sodium deoxycholate, 100 μg/ml phenylmethylsulfonyl-fluoride, complete protease inhibitor cocktail tablet [Boehringer Mannheim Biochemicals] [1 tablet/50 ml of buffer]). .. Radioactivity in 5-μl samples of lysates was quantified in a Beckman scintillation counter (model LS5000TD).

Lysis:

Article Title: Human Cytomegalovirus Inhibits Transcription of the CC Chemokine MCP-1 Gene
Article Snippet: Medium was collected from infected HFF cultures, and the supernatant was assayed by enzyme-linked immunosorbent assay (ELISA), using a Quantikine plate specific for human MCP-1 (R & D Systems) in accordance with the manufacturer’s protocol. .. For immunoprecipitations, confluent 10-cm-diameter dishes of HFFs were labeled with [35 S]methionine for 1 h and lysed in 0.5 ml of lysis buffer (50 mM Tris-Cl [pH 8.0], 150 mM NaCl, 0.1% sodium dodecyl sulfate, 1.0% Nonidet P-40, 0.5% sodium deoxycholate, 100 μg/ml phenylmethylsulfonyl-fluoride, complete protease inhibitor cocktail tablet [Boehringer Mannheim Biochemicals] [1 tablet/50 ml of buffer]). .. Radioactivity in 5-μl samples of lysates was quantified in a Beckman scintillation counter (model LS5000TD).

Article Title: Effect of Brain- and Tumor-Derived Connective Tissue Growth Factor on Glioma Invasion
Article Snippet: To examine whether ITGB1 is needed for TrkA activation in the presence of CTGF, 0827 TIC/TSCs or 0827 TIC/TSCs with stable knockdown of TrkA or ITGB1 were seeded into 10-cm plates (1 × 106 cells per plate, two plates per condition) for 12 hours and incubated for 1.5 hours with CTGF (200 ng/mL). .. TIC/TSCs were then harvested with ice-cold lysis buffer (50 mM Tris, 150 mM NaCl, 2.5 mM EDTA, 0.1% sodium dodecyl sulfate [SDS], 0.5% sodium deoxycholate, 1% Nonidet P-40, and 0.02% sodium azide) containing Complete-Mini protease inhibitor (one tablet per 10 mL; Boehringer Mannheim GmBH, Mannheim, Germany). .. Protein content in the lysates was determined using a detergent-compatible Bradford protein assay (Bio-Rad Laboratories, Hercules, CA).

Article Title: PR48, a Novel Regulatory Subunit of Protein Phosphatase 2A, Interacts with Cdc6 and Modulates DNA Replication in Human Cells
Article Snippet: Coimmunoprecipitation of transiently expressed Cdc6–c-Myc and HA-PR48 in mammalian cells was performed according to the instructions of the manufacturer of the anti–c-Myc antibody (Boehringer Mannheim). .. Briefly, 24 h after transfection, HeLa or C2C12 cells were incubated on ice for 20 min in lysis buffer containing 20 mM Tris-HCl (pH 7.5), 0.2% Nonidet P-40, 20% glycerol, 200 mM NaCl, 1 mM EDTA, 1 mM EGTA, and a protease inhibitor cocktail (Boehringer Mannheim). .. Lysates were centrifuged at 14,000 × g for 10 min, and protein complexes were immunoprecipitated from the supernatant by the addition of 4 μg of a murine monoclonal antibody that recognizes the c-Myc epitope (Boehringer Mannheim) and incubation for 1 h with rotation at 4°C.

Article Title: Strain Differences in Behavioral and Cellular Responses to Perinatal Hypoxia and Relationships to Neural Stem Cell Survival and Self-Renewal
Article Snippet: PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories) in a final volume of 25 μl, starting with a 3-minute template denaturation step at 95°C followed by 40 cycles of 95°C for 30 seconds and 55°C for 30 seconds. .. NSCs were homogenized in lysis buffer composed of 50 mmol/L Tris-HCl, pH 7.4, 150 mmol/L NaCl, 1% Nonidet P-40, 10% glycerol, 1 mmol/L sodium orthovanadate, 1 mmol/L phenylmethylsulfonyl fluoride, and protease inhibitor cocktail (Boehringer Mannheim GmbH, Mannheim, Germany). ..

Protease Inhibitor:

Article Title: Human Cytomegalovirus Inhibits Transcription of the CC Chemokine MCP-1 Gene
Article Snippet: Medium was collected from infected HFF cultures, and the supernatant was assayed by enzyme-linked immunosorbent assay (ELISA), using a Quantikine plate specific for human MCP-1 (R & D Systems) in accordance with the manufacturer’s protocol. .. For immunoprecipitations, confluent 10-cm-diameter dishes of HFFs were labeled with [35 S]methionine for 1 h and lysed in 0.5 ml of lysis buffer (50 mM Tris-Cl [pH 8.0], 150 mM NaCl, 0.1% sodium dodecyl sulfate, 1.0% Nonidet P-40, 0.5% sodium deoxycholate, 100 μg/ml phenylmethylsulfonyl-fluoride, complete protease inhibitor cocktail tablet [Boehringer Mannheim Biochemicals] [1 tablet/50 ml of buffer]). .. Radioactivity in 5-μl samples of lysates was quantified in a Beckman scintillation counter (model LS5000TD).

Article Title: Functional communication between endogenous BRCA1 and its partner, BARD1, during Xenopus laevis development
Article Snippet: .. Cells were lysed in a buffer containing 100 mM Hepes (pH 7.5), 200 mM NaCl, 40 mM EDTA, 4 mM EGTA, 100 mM NaF, 20 mM β-glycerophosphate, 2 mM sodium orthovanadate, 1% Nonidet P-40, and 1 tablet per 50 ml of the Complete Protease Inhibitor mixture (Boehringer Mannheim). .. For immunoprecipitations (IPs), extracts from 293 T cells or from 8–10 eggs or embryos were incubated with 1 μg of Ab on ice for 2–3 h, followed by incubation with 20 μl of protein A- or protein G-Sepharose beads (Amersham Pharmacia).

Article Title: Effect of Brain- and Tumor-Derived Connective Tissue Growth Factor on Glioma Invasion
Article Snippet: To examine whether ITGB1 is needed for TrkA activation in the presence of CTGF, 0827 TIC/TSCs or 0827 TIC/TSCs with stable knockdown of TrkA or ITGB1 were seeded into 10-cm plates (1 × 106 cells per plate, two plates per condition) for 12 hours and incubated for 1.5 hours with CTGF (200 ng/mL). .. TIC/TSCs were then harvested with ice-cold lysis buffer (50 mM Tris, 150 mM NaCl, 2.5 mM EDTA, 0.1% sodium dodecyl sulfate [SDS], 0.5% sodium deoxycholate, 1% Nonidet P-40, and 0.02% sodium azide) containing Complete-Mini protease inhibitor (one tablet per 10 mL; Boehringer Mannheim GmBH, Mannheim, Germany). .. Protein content in the lysates was determined using a detergent-compatible Bradford protein assay (Bio-Rad Laboratories, Hercules, CA).

Article Title: PR48, a Novel Regulatory Subunit of Protein Phosphatase 2A, Interacts with Cdc6 and Modulates DNA Replication in Human Cells
Article Snippet: Coimmunoprecipitation of transiently expressed Cdc6–c-Myc and HA-PR48 in mammalian cells was performed according to the instructions of the manufacturer of the anti–c-Myc antibody (Boehringer Mannheim). .. Briefly, 24 h after transfection, HeLa or C2C12 cells were incubated on ice for 20 min in lysis buffer containing 20 mM Tris-HCl (pH 7.5), 0.2% Nonidet P-40, 20% glycerol, 200 mM NaCl, 1 mM EDTA, 1 mM EGTA, and a protease inhibitor cocktail (Boehringer Mannheim). .. Lysates were centrifuged at 14,000 × g for 10 min, and protein complexes were immunoprecipitated from the supernatant by the addition of 4 μg of a murine monoclonal antibody that recognizes the c-Myc epitope (Boehringer Mannheim) and incubation for 1 h with rotation at 4°C.

Article Title: Strain Differences in Behavioral and Cellular Responses to Perinatal Hypoxia and Relationships to Neural Stem Cell Survival and Self-Renewal
Article Snippet: PCR was performed using iQ SYBR Green Supermix (Bio-Rad Laboratories) in a final volume of 25 μl, starting with a 3-minute template denaturation step at 95°C followed by 40 cycles of 95°C for 30 seconds and 55°C for 30 seconds. .. NSCs were homogenized in lysis buffer composed of 50 mmol/L Tris-HCl, pH 7.4, 150 mmol/L NaCl, 1% Nonidet P-40, 10% glycerol, 1 mmol/L sodium orthovanadate, 1 mmol/L phenylmethylsulfonyl fluoride, and protease inhibitor cocktail (Boehringer Mannheim GmbH, Mannheim, Germany). ..

Transfection:

Article Title: PR48, a Novel Regulatory Subunit of Protein Phosphatase 2A, Interacts with Cdc6 and Modulates DNA Replication in Human Cells
Article Snippet: Coimmunoprecipitation of transiently expressed Cdc6–c-Myc and HA-PR48 in mammalian cells was performed according to the instructions of the manufacturer of the anti–c-Myc antibody (Boehringer Mannheim). .. Briefly, 24 h after transfection, HeLa or C2C12 cells were incubated on ice for 20 min in lysis buffer containing 20 mM Tris-HCl (pH 7.5), 0.2% Nonidet P-40, 20% glycerol, 200 mM NaCl, 1 mM EDTA, 1 mM EGTA, and a protease inhibitor cocktail (Boehringer Mannheim). .. Lysates were centrifuged at 14,000 × g for 10 min, and protein complexes were immunoprecipitated from the supernatant by the addition of 4 μg of a murine monoclonal antibody that recognizes the c-Myc epitope (Boehringer Mannheim) and incubation for 1 h with rotation at 4°C.

Incubation:

Article Title: PR48, a Novel Regulatory Subunit of Protein Phosphatase 2A, Interacts with Cdc6 and Modulates DNA Replication in Human Cells
Article Snippet: Coimmunoprecipitation of transiently expressed Cdc6–c-Myc and HA-PR48 in mammalian cells was performed according to the instructions of the manufacturer of the anti–c-Myc antibody (Boehringer Mannheim). .. Briefly, 24 h after transfection, HeLa or C2C12 cells were incubated on ice for 20 min in lysis buffer containing 20 mM Tris-HCl (pH 7.5), 0.2% Nonidet P-40, 20% glycerol, 200 mM NaCl, 1 mM EDTA, 1 mM EGTA, and a protease inhibitor cocktail (Boehringer Mannheim). .. Lysates were centrifuged at 14,000 × g for 10 min, and protein complexes were immunoprecipitated from the supernatant by the addition of 4 μg of a murine monoclonal antibody that recognizes the c-Myc epitope (Boehringer Mannheim) and incubation for 1 h with rotation at 4°C.

Article Title: Internalization of CD26 by mannose 6-phosphate/insulin-like growth factor II receptor contributes to T cell activation
Article Snippet: Protein sequencing was carried out by the Harvard Microchemistry Facility. .. For glycosidase or phosphatase treatment, sCD26 was denatured by boiling with SDS and incubated for 12 h at 37°C in incubation buffer [20 mM Tris-malate (pH 7.0 or pH 8.0)/1% Nonidet P-40] containing 10 microunits/μl N -glycosidase-F and 25 microunits/μl O -glycosidase (Boehringer Mannheim) (pH 7.0) or 50 milliunits/μl Escherichia coli alkaline phosphatase (Sigma) (pH 8.0). ..

Centrifugation:

Article Title: Purine Salvage in Two Halophilic Archaea: Characterization of Salvage Pathways and Isolation of Mutants Resistant to Purine Analogs
Article Snippet: .. Cells in 40 ml of culture were harvested by centrifugation and resuspended in 0.5 ml of buffer containing 100 mM Tris-HCl (pH 7.5), 3.5 M KCl, 0.2% Nonidet P-40, and 15 units of DNase I (Boehringer, Mannheim, Germany). .. When no KCl was added to the buffer, the salt concentration of the crude extract was found by conductivity measurements to be equivalent to 0.25 M KCl.

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    Boehringer Mannheim immunoprecipitation ip buffer
    Full-length BCL-6 and BCoR interact in vitro and in vivo, the POZ domain of BCL-6 is both necessary and sufficient for interaction with BCoR in vitro, and BCL-6 and BCoR colocalize in the nucleus. ( A ) Structure of the proteins used in B and C . (Solid oval) The POZ domain; (solid rectangle) zinc fingers. (*) The myc epitope tag. ( B ) Coimmunoprecipitation of BCL-6 and BCoR in vitro. Pair-wise combinations of [ 35 S]methionine-labeled nontagged BCL-6 derivatives and myc-tagged BCoR were produced by cotranslation and immunoprecipitated with the α-myc antibody. The immune complexes were recovered and analyzed by SDS-PAGE (10% for lanes 1–5; 17% for lanes 6,7 ). The intensity of myc–BCoR in lanes 6 and 7 appears stronger due to compression on the 17% gel of non-full-length myc-BCoR proteins. Input represents 20% of the total used. (●) The expected migration of the nontagged BCL-6 derivatives. ( C ) Coimmunoprecipitation of BCL-6 and BCoR in vivo. 293 cells were transfected with 3 μg of BCL-6 expression plasmid alone (lane 2 ) or cotransfected with 3 μg each of BCL-6 and myc-tagged BCoR expression plasmids (lanes 1,3 ). Cell lysates were immunoprecipitated with α-myc antibody (lanes 2,3 ) and the recovered proteins were detected by Western blot analysis using N-3 α-BCL-6 and α-myc antibodies. The input (lane 1 ) corresponds to 1/50 of the lysate used in the <t>immunoprecipitation</t> (lane 3 ). ( D ) Subcellular localization of BCoR and BCoR-S. Immunofluorescence was performed on HeLa cells transfected with 1.2 μg of either myc-tagged BCoR or BCoR-S expression plasmids. Multiple Z series confocal microscopy images of the given fields were compiled for each image of BCoR and BCoR-S. ( E ) Colocalization of BCL-6 and BCoR. Coimmunofluorescence was performed on HeLa cells cotransfected with 0.6 μg each of BCL-6 and myc-tagged BCoR, and the data collected as in D . The compiled images of BCoR and BCL-6 were merged to detect colocalization.
    Immunoprecipitation Ip Buffer, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation ip buffer/product/Boehringer Mannheim
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitation ip buffer - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    86
    Boehringer Mannheim immunoprecipitation assay buffer
    Molecular characterization of the calsenilin knock-out mouse. A , Gene targeting strategy. Top, Wild-type calsenilin locus showing exon 2 and the position of the 5′ and 3′ probes used for genotyping. Bottom, Mutant locus in which exon 2 has been replaced with an IRES, the marker gene β -gal , and the neomycin (neo) resistance gene, the latter under the control of the MC1 promoter. B , Southern blot analysis of genomic DNA from calsenilin +/+, +/-, and -/- mice. Left, Hind III digest; right, Nco I digest. C , Northern blot analysis of mRNA extracted from the brains of calsenilin +/+, +/-, and -/- mice. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is included as a loading control. D ). The asterisk indicates a band migrating just above mouse calsenilin, and the position of IgG (from the <t>immunoprecipitation)</t> is also indicated.
    Immunoprecipitation Assay Buffer, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoprecipitation assay buffer/product/Boehringer Mannheim
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immunoprecipitation assay buffer - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    86
    Boehringer Mannheim nonidet p 40
    Molecular characterization of the calsenilin knock-out mouse. A , Gene targeting strategy. Top, Wild-type calsenilin locus showing exon 2 and the position of the 5′ and 3′ probes used for genotyping. Bottom, Mutant locus in which exon 2 has been replaced with an IRES, the marker gene β -gal , and the neomycin (neo) resistance gene, the latter under the control of the MC1 promoter. B , Southern blot analysis of genomic DNA from calsenilin +/+, +/-, and -/- mice. Left, Hind III digest; right, Nco I digest. C , Northern blot analysis of mRNA extracted from the brains of calsenilin +/+, +/-, and -/- mice. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is included as a loading control. D ). The asterisk indicates a band migrating just above mouse calsenilin, and the position of IgG (from the <t>immunoprecipitation)</t> is also indicated.
    Nonidet P 40, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nonidet p 40/product/Boehringer Mannheim
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nonidet p 40 - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    Image Search Results


    Full-length BCL-6 and BCoR interact in vitro and in vivo, the POZ domain of BCL-6 is both necessary and sufficient for interaction with BCoR in vitro, and BCL-6 and BCoR colocalize in the nucleus. ( A ) Structure of the proteins used in B and C . (Solid oval) The POZ domain; (solid rectangle) zinc fingers. (*) The myc epitope tag. ( B ) Coimmunoprecipitation of BCL-6 and BCoR in vitro. Pair-wise combinations of [ 35 S]methionine-labeled nontagged BCL-6 derivatives and myc-tagged BCoR were produced by cotranslation and immunoprecipitated with the α-myc antibody. The immune complexes were recovered and analyzed by SDS-PAGE (10% for lanes 1–5; 17% for lanes 6,7 ). The intensity of myc–BCoR in lanes 6 and 7 appears stronger due to compression on the 17% gel of non-full-length myc-BCoR proteins. Input represents 20% of the total used. (●) The expected migration of the nontagged BCL-6 derivatives. ( C ) Coimmunoprecipitation of BCL-6 and BCoR in vivo. 293 cells were transfected with 3 μg of BCL-6 expression plasmid alone (lane 2 ) or cotransfected with 3 μg each of BCL-6 and myc-tagged BCoR expression plasmids (lanes 1,3 ). Cell lysates were immunoprecipitated with α-myc antibody (lanes 2,3 ) and the recovered proteins were detected by Western blot analysis using N-3 α-BCL-6 and α-myc antibodies. The input (lane 1 ) corresponds to 1/50 of the lysate used in the immunoprecipitation (lane 3 ). ( D ) Subcellular localization of BCoR and BCoR-S. Immunofluorescence was performed on HeLa cells transfected with 1.2 μg of either myc-tagged BCoR or BCoR-S expression plasmids. Multiple Z series confocal microscopy images of the given fields were compiled for each image of BCoR and BCoR-S. ( E ) Colocalization of BCL-6 and BCoR. Coimmunofluorescence was performed on HeLa cells cotransfected with 0.6 μg each of BCL-6 and myc-tagged BCoR, and the data collected as in D . The compiled images of BCoR and BCL-6 were merged to detect colocalization.

    Journal: Genes & Development

    Article Title: BCoR, a novel corepressor involved in BCL-6 repression

    doi:

    Figure Lengend Snippet: Full-length BCL-6 and BCoR interact in vitro and in vivo, the POZ domain of BCL-6 is both necessary and sufficient for interaction with BCoR in vitro, and BCL-6 and BCoR colocalize in the nucleus. ( A ) Structure of the proteins used in B and C . (Solid oval) The POZ domain; (solid rectangle) zinc fingers. (*) The myc epitope tag. ( B ) Coimmunoprecipitation of BCL-6 and BCoR in vitro. Pair-wise combinations of [ 35 S]methionine-labeled nontagged BCL-6 derivatives and myc-tagged BCoR were produced by cotranslation and immunoprecipitated with the α-myc antibody. The immune complexes were recovered and analyzed by SDS-PAGE (10% for lanes 1–5; 17% for lanes 6,7 ). The intensity of myc–BCoR in lanes 6 and 7 appears stronger due to compression on the 17% gel of non-full-length myc-BCoR proteins. Input represents 20% of the total used. (●) The expected migration of the nontagged BCL-6 derivatives. ( C ) Coimmunoprecipitation of BCL-6 and BCoR in vivo. 293 cells were transfected with 3 μg of BCL-6 expression plasmid alone (lane 2 ) or cotransfected with 3 μg each of BCL-6 and myc-tagged BCoR expression plasmids (lanes 1,3 ). Cell lysates were immunoprecipitated with α-myc antibody (lanes 2,3 ) and the recovered proteins were detected by Western blot analysis using N-3 α-BCL-6 and α-myc antibodies. The input (lane 1 ) corresponds to 1/50 of the lysate used in the immunoprecipitation (lane 3 ). ( D ) Subcellular localization of BCoR and BCoR-S. Immunofluorescence was performed on HeLa cells transfected with 1.2 μg of either myc-tagged BCoR or BCoR-S expression plasmids. Multiple Z series confocal microscopy images of the given fields were compiled for each image of BCoR and BCoR-S. ( E ) Colocalization of BCL-6 and BCoR. Coimmunofluorescence was performed on HeLa cells cotransfected with 0.6 μg each of BCL-6 and myc-tagged BCoR, and the data collected as in D . The compiled images of BCoR and BCL-6 were merged to detect colocalization.

    Article Snippet: Transfected cells were washed twice with PBS and lysed in 500 μl of cold immunoprecipitation (IP) buffer (PBS, 10% glycerol, 0.5% NP-40, and a complete protease inhibitor cocktail from Boehringer-Mannheim) for 10 min on ice.

    Techniques: In Vitro, In Vivo, Zinc-Fingers, Labeling, Produced, Immunoprecipitation, SDS Page, Migration, Transfection, Expressing, Plasmid Preparation, Western Blot, Immunofluorescence, Confocal Microscopy

    Molecular characterization of the calsenilin knock-out mouse. A , Gene targeting strategy. Top, Wild-type calsenilin locus showing exon 2 and the position of the 5′ and 3′ probes used for genotyping. Bottom, Mutant locus in which exon 2 has been replaced with an IRES, the marker gene β -gal , and the neomycin (neo) resistance gene, the latter under the control of the MC1 promoter. B , Southern blot analysis of genomic DNA from calsenilin +/+, +/-, and -/- mice. Left, Hind III digest; right, Nco I digest. C , Northern blot analysis of mRNA extracted from the brains of calsenilin +/+, +/-, and -/- mice. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is included as a loading control. D ). The asterisk indicates a band migrating just above mouse calsenilin, and the position of IgG (from the immunoprecipitation) is also indicated.

    Journal: The Journal of Neuroscience

    Article Title: Altered Aβ Formation and Long-Term Potentiation in a Calsenilin Knock-Out

    doi: 10.1523/JNEUROSCI.23-27-09097.2003

    Figure Lengend Snippet: Molecular characterization of the calsenilin knock-out mouse. A , Gene targeting strategy. Top, Wild-type calsenilin locus showing exon 2 and the position of the 5′ and 3′ probes used for genotyping. Bottom, Mutant locus in which exon 2 has been replaced with an IRES, the marker gene β -gal , and the neomycin (neo) resistance gene, the latter under the control of the MC1 promoter. B , Southern blot analysis of genomic DNA from calsenilin +/+, +/-, and -/- mice. Left, Hind III digest; right, Nco I digest. C , Northern blot analysis of mRNA extracted from the brains of calsenilin +/+, +/-, and -/- mice. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is included as a loading control. D ). The asterisk indicates a band migrating just above mouse calsenilin, and the position of IgG (from the immunoprecipitation) is also indicated.

    Article Snippet: Cortex, cerebellum, and hippocampus from wild-type and calsenilin knock-out mice were homogenized in immunoprecipitation assay buffer (50 m m Tris, pH 7.4, 150 m m NaCl, 2 m m EDTA, 1% Triton X-100, 1% NP-40, and 0.25% sodium deoxycholate) with protease inhibitors (Complete 1; Boehringer Mannheim, Indianapolis, IN).

    Techniques: Knock-Out, Mutagenesis, Marker, Southern Blot, Mouse Assay, Northern Blot, Immunoprecipitation