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GE Healthcare complementary dna cdna
Complementary Dna Cdna, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/complementary dna cdna/product/GE Healthcare
Average 92 stars, based on 4 article reviews
Price from $9.99 to $1999.99
complementary dna cdna - by Bioz Stars, 2020-08
92/100 stars

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Enzyme-linked Immunosorbent Assay:

Article Title: Generation of an alpaca-derived nanobody recognizing γ-H2AX
Article Snippet: .. Bound alpaca antibodies were further detected with HRP-conjugated anti-alpaca IgG antibody (Bethyl Laboratories Inc, Montgomery, Alabama, USA). (3) Upon positive ELISA test, B cells were isolated with a Ficoll gradient using UNI-SEPMAXI (Novamed Ltd., Jerusalem, Israel). (4) From the B cells, RNA was extracted with the TRIzol reagent (Life Technologies, Carlsbad, California, USA) according to the manufacturer’s protocol. (5) From this RNA, complementary DNA (cDNA) was generated using the First-Strand cDNA Synthesis Kit (GE Healthcare, Uppsala, Sweden) according to the manufacturer’s protocol. (6) VHHs were amplified by three sequential PCR reactions. cDNA was used as the DNA template for the first PCR. .. For the PCR reactions, the following primers were used: 1st PCR: Forward primer CALL001: 5′-GTC CTG GCT GCT CTT CTA CA A GG-3′ Reverse primer CALL002: 5′-GGT ACG TGC TGT TGA ACT GTT CC-3′; 2nd PCR: Forward primer SM017: 5′-CCA GCC GGC CAT GGC TCA GGT GCA GCT GGT GGA GTC TGG-3′ Reverse primer SM018: 5′-CCA GCC GGC CAT GGC TGA TGT GCA GCT GGT GGA GTC TGG-3′; 3rd PCR: Forward primer A4short: 5′-CAT GCC ATG ACT CGC GGC CAC GCC GGC CAT GGC-3′ Reverse primer 38: 5′-GGA CTA GTG CGG CCG CTG GAG ACG GTG ACC TGG GT-3′. (7) The amplified product and the plasmid vector pHEN4 were digested with NotI and NcoI restriction enzymes, thus producing compatible overhangs to ligate. (8) Electro-competent TG1 cells (Agilent Technologies GmbH & Co.KG, Waldbronn, Baden-Wuerttemberg, Germany) were used to generate VHH libraries.

Amplification:

Article Title: Generation of an alpaca-derived nanobody recognizing γ-H2AX
Article Snippet: .. Bound alpaca antibodies were further detected with HRP-conjugated anti-alpaca IgG antibody (Bethyl Laboratories Inc, Montgomery, Alabama, USA). (3) Upon positive ELISA test, B cells were isolated with a Ficoll gradient using UNI-SEPMAXI (Novamed Ltd., Jerusalem, Israel). (4) From the B cells, RNA was extracted with the TRIzol reagent (Life Technologies, Carlsbad, California, USA) according to the manufacturer’s protocol. (5) From this RNA, complementary DNA (cDNA) was generated using the First-Strand cDNA Synthesis Kit (GE Healthcare, Uppsala, Sweden) according to the manufacturer’s protocol. (6) VHHs were amplified by three sequential PCR reactions. cDNA was used as the DNA template for the first PCR. .. For the PCR reactions, the following primers were used: 1st PCR: Forward primer CALL001: 5′-GTC CTG GCT GCT CTT CTA CA A GG-3′ Reverse primer CALL002: 5′-GGT ACG TGC TGT TGA ACT GTT CC-3′; 2nd PCR: Forward primer SM017: 5′-CCA GCC GGC CAT GGC TCA GGT GCA GCT GGT GGA GTC TGG-3′ Reverse primer SM018: 5′-CCA GCC GGC CAT GGC TGA TGT GCA GCT GGT GGA GTC TGG-3′; 3rd PCR: Forward primer A4short: 5′-CAT GCC ATG ACT CGC GGC CAC GCC GGC CAT GGC-3′ Reverse primer 38: 5′-GGA CTA GTG CGG CCG CTG GAG ACG GTG ACC TGG GT-3′. (7) The amplified product and the plasmid vector pHEN4 were digested with NotI and NcoI restriction enzymes, thus producing compatible overhangs to ligate. (8) Electro-competent TG1 cells (Agilent Technologies GmbH & Co.KG, Waldbronn, Baden-Wuerttemberg, Germany) were used to generate VHH libraries.

Synthesized:

Article Title: MYC-dependent downregulation of telomerase by FLT3 inhibitors is required for their therapeutic efficacy on acute myeloid leukemia
Article Snippet: .. Complementary DNA (cDNA) was synthesized using random primers (N6) (Amersham, Buckinghamshire, UK) and M-MLV reverse transcriptase. .. The PCR primers are listed in Table . β2-Microglobulin (β2-M) expression was used as a control for RNA loading and RT efficiency and amplified in parallel. qPCR was carried out in an ABI7700 sequence detector (Applied Biosystems, Foster City, CA) using a SYBR Green kit (Applied Biosystems) with triplicates.

Isolation:

Article Title: Generation of an alpaca-derived nanobody recognizing γ-H2AX
Article Snippet: .. Bound alpaca antibodies were further detected with HRP-conjugated anti-alpaca IgG antibody (Bethyl Laboratories Inc, Montgomery, Alabama, USA). (3) Upon positive ELISA test, B cells were isolated with a Ficoll gradient using UNI-SEPMAXI (Novamed Ltd., Jerusalem, Israel). (4) From the B cells, RNA was extracted with the TRIzol reagent (Life Technologies, Carlsbad, California, USA) according to the manufacturer’s protocol. (5) From this RNA, complementary DNA (cDNA) was generated using the First-Strand cDNA Synthesis Kit (GE Healthcare, Uppsala, Sweden) according to the manufacturer’s protocol. (6) VHHs were amplified by three sequential PCR reactions. cDNA was used as the DNA template for the first PCR. .. For the PCR reactions, the following primers were used: 1st PCR: Forward primer CALL001: 5′-GTC CTG GCT GCT CTT CTA CA A GG-3′ Reverse primer CALL002: 5′-GGT ACG TGC TGT TGA ACT GTT CC-3′; 2nd PCR: Forward primer SM017: 5′-CCA GCC GGC CAT GGC TCA GGT GCA GCT GGT GGA GTC TGG-3′ Reverse primer SM018: 5′-CCA GCC GGC CAT GGC TGA TGT GCA GCT GGT GGA GTC TGG-3′; 3rd PCR: Forward primer A4short: 5′-CAT GCC ATG ACT CGC GGC CAC GCC GGC CAT GGC-3′ Reverse primer 38: 5′-GGA CTA GTG CGG CCG CTG GAG ACG GTG ACC TGG GT-3′. (7) The amplified product and the plasmid vector pHEN4 were digested with NotI and NcoI restriction enzymes, thus producing compatible overhangs to ligate. (8) Electro-competent TG1 cells (Agilent Technologies GmbH & Co.KG, Waldbronn, Baden-Wuerttemberg, Germany) were used to generate VHH libraries.

Generated:

Article Title: Generation of an alpaca-derived nanobody recognizing γ-H2AX
Article Snippet: .. Bound alpaca antibodies were further detected with HRP-conjugated anti-alpaca IgG antibody (Bethyl Laboratories Inc, Montgomery, Alabama, USA). (3) Upon positive ELISA test, B cells were isolated with a Ficoll gradient using UNI-SEPMAXI (Novamed Ltd., Jerusalem, Israel). (4) From the B cells, RNA was extracted with the TRIzol reagent (Life Technologies, Carlsbad, California, USA) according to the manufacturer’s protocol. (5) From this RNA, complementary DNA (cDNA) was generated using the First-Strand cDNA Synthesis Kit (GE Healthcare, Uppsala, Sweden) according to the manufacturer’s protocol. (6) VHHs were amplified by three sequential PCR reactions. cDNA was used as the DNA template for the first PCR. .. For the PCR reactions, the following primers were used: 1st PCR: Forward primer CALL001: 5′-GTC CTG GCT GCT CTT CTA CA A GG-3′ Reverse primer CALL002: 5′-GGT ACG TGC TGT TGA ACT GTT CC-3′; 2nd PCR: Forward primer SM017: 5′-CCA GCC GGC CAT GGC TCA GGT GCA GCT GGT GGA GTC TGG-3′ Reverse primer SM018: 5′-CCA GCC GGC CAT GGC TGA TGT GCA GCT GGT GGA GTC TGG-3′; 3rd PCR: Forward primer A4short: 5′-CAT GCC ATG ACT CGC GGC CAC GCC GGC CAT GGC-3′ Reverse primer 38: 5′-GGA CTA GTG CGG CCG CTG GAG ACG GTG ACC TGG GT-3′. (7) The amplified product and the plasmid vector pHEN4 were digested with NotI and NcoI restriction enzymes, thus producing compatible overhangs to ligate. (8) Electro-competent TG1 cells (Agilent Technologies GmbH & Co.KG, Waldbronn, Baden-Wuerttemberg, Germany) were used to generate VHH libraries.

Real-time Polymerase Chain Reaction:

Article Title: Effects of Isoxazolo-Pyridinone 7e, a Potent Activator of the Nurr1 Signaling Pathway, on Experimental Autoimmune Encephalomyelitis in Mice
Article Snippet: .. Total RNA was reverse-transcribed to complementary DNA (cDNA) using the Ready-To-Go You-Prime First-Strand Beads (Amersham, Airlington Heights, IL, USA) and Random Hexamer (New England Biolabs, Ipswich, MA, USA) according to the manufacturer's instructions. cDNA was used as a template for a real-time PCR analysis. .. Quantitative Real-time PCR was carried out using the ABI Prism 7000 Sequence Detection System (Applied Biosystems, Life Technology, Paisley, UK).

Sequencing:

Article Title: The N-terminal GTPase domain of p190RhoGAP proteins is a pseudoGTPase
Article Snippet: .. The complementary DNA (cDNA) encoding full-length Rattus norvegicus (rat) p190RhoGAP-A (ARHGAP35) protein (NCBI Reference Sequence: , UniProt: A0A0G2KB46) was used as a PCR-template to amplify the segment encoding residues 1-266 which was inserted into pGEX-6p1 vector (GE Healthcare) for expression as a GST-fusion protein in Escherichia coli . .. A shorter N-GTPase construct of residues 13-249 was inserted into a modified pET vector for expression as a His-tagged protein.

Random Hexamer Labeling:

Article Title: Effects of Isoxazolo-Pyridinone 7e, a Potent Activator of the Nurr1 Signaling Pathway, on Experimental Autoimmune Encephalomyelitis in Mice
Article Snippet: .. Total RNA was reverse-transcribed to complementary DNA (cDNA) using the Ready-To-Go You-Prime First-Strand Beads (Amersham, Airlington Heights, IL, USA) and Random Hexamer (New England Biolabs, Ipswich, MA, USA) according to the manufacturer's instructions. cDNA was used as a template for a real-time PCR analysis. .. Quantitative Real-time PCR was carried out using the ABI Prism 7000 Sequence Detection System (Applied Biosystems, Life Technology, Paisley, UK).

Expressing:

Article Title: The N-terminal GTPase domain of p190RhoGAP proteins is a pseudoGTPase
Article Snippet: .. The complementary DNA (cDNA) encoding full-length Rattus norvegicus (rat) p190RhoGAP-A (ARHGAP35) protein (NCBI Reference Sequence: , UniProt: A0A0G2KB46) was used as a PCR-template to amplify the segment encoding residues 1-266 which was inserted into pGEX-6p1 vector (GE Healthcare) for expression as a GST-fusion protein in Escherichia coli . .. A shorter N-GTPase construct of residues 13-249 was inserted into a modified pET vector for expression as a His-tagged protein.

Polymerase Chain Reaction:

Article Title: Generation of an alpaca-derived nanobody recognizing γ-H2AX
Article Snippet: .. Bound alpaca antibodies were further detected with HRP-conjugated anti-alpaca IgG antibody (Bethyl Laboratories Inc, Montgomery, Alabama, USA). (3) Upon positive ELISA test, B cells were isolated with a Ficoll gradient using UNI-SEPMAXI (Novamed Ltd., Jerusalem, Israel). (4) From the B cells, RNA was extracted with the TRIzol reagent (Life Technologies, Carlsbad, California, USA) according to the manufacturer’s protocol. (5) From this RNA, complementary DNA (cDNA) was generated using the First-Strand cDNA Synthesis Kit (GE Healthcare, Uppsala, Sweden) according to the manufacturer’s protocol. (6) VHHs were amplified by three sequential PCR reactions. cDNA was used as the DNA template for the first PCR. .. For the PCR reactions, the following primers were used: 1st PCR: Forward primer CALL001: 5′-GTC CTG GCT GCT CTT CTA CA A GG-3′ Reverse primer CALL002: 5′-GGT ACG TGC TGT TGA ACT GTT CC-3′; 2nd PCR: Forward primer SM017: 5′-CCA GCC GGC CAT GGC TCA GGT GCA GCT GGT GGA GTC TGG-3′ Reverse primer SM018: 5′-CCA GCC GGC CAT GGC TGA TGT GCA GCT GGT GGA GTC TGG-3′; 3rd PCR: Forward primer A4short: 5′-CAT GCC ATG ACT CGC GGC CAC GCC GGC CAT GGC-3′ Reverse primer 38: 5′-GGA CTA GTG CGG CCG CTG GAG ACG GTG ACC TGG GT-3′. (7) The amplified product and the plasmid vector pHEN4 were digested with NotI and NcoI restriction enzymes, thus producing compatible overhangs to ligate. (8) Electro-competent TG1 cells (Agilent Technologies GmbH & Co.KG, Waldbronn, Baden-Wuerttemberg, Germany) were used to generate VHH libraries.

Article Title: The N-terminal GTPase domain of p190RhoGAP proteins is a pseudoGTPase
Article Snippet: .. The complementary DNA (cDNA) encoding full-length Rattus norvegicus (rat) p190RhoGAP-A (ARHGAP35) protein (NCBI Reference Sequence: , UniProt: A0A0G2KB46) was used as a PCR-template to amplify the segment encoding residues 1-266 which was inserted into pGEX-6p1 vector (GE Healthcare) for expression as a GST-fusion protein in Escherichia coli . .. A shorter N-GTPase construct of residues 13-249 was inserted into a modified pET vector for expression as a His-tagged protein.

Plasmid Preparation:

Article Title: The N-terminal GTPase domain of p190RhoGAP proteins is a pseudoGTPase
Article Snippet: .. The complementary DNA (cDNA) encoding full-length Rattus norvegicus (rat) p190RhoGAP-A (ARHGAP35) protein (NCBI Reference Sequence: , UniProt: A0A0G2KB46) was used as a PCR-template to amplify the segment encoding residues 1-266 which was inserted into pGEX-6p1 vector (GE Healthcare) for expression as a GST-fusion protein in Escherichia coli . .. A shorter N-GTPase construct of residues 13-249 was inserted into a modified pET vector for expression as a His-tagged protein.

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  • 89
    GE Healthcare pcr amplified cdna fragments
    Characterization of androgen-responsive non-coding intronic RNAs . Selected intronic transcripts extended by <t>RACE-PCR</t> (A-D, red boxes) or gene-specific PCR (E, yellow box) are shown. Extended unspliced antisense intronic transcript fragments were mapped to the genomic sequence (A-E, gray lines) relative to the respective spliced protein-coding transcript expressed in the opposite strand (A-E, green arrows). Double-stranded <t>cDNA</t> clones spotted on the microarrays are represented as blue boxes (A-E). A previously described antisense unspliced intronic transcript mapping to GAS6 locus is shown (E, black arrow). Chromosome coordinates and the length of extended intronic transcript fragments are indicated. (F) Strand-specific multi-tissue northern blot using probes complementary to the antisense intronic transcript mapped in the GAS6 locus.
    Pcr Amplified Cdna Fragments, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr amplified cdna fragments/product/GE Healthcare
    Average 89 stars, based on 1 article reviews
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    92
    GE Healthcare strand cdna synthesis
    Northern blot with total Cuscuta <t>RNA</t> . Cuscuta RNA from shoots (shoot) or shoot material with haustoria (haustoria). Upper panel: hybridization signal with a <t>cuscutain-cDNA</t> probe. Lower panel: Ethidium bromide stained total RNA to indicate even loading.
    Strand Cdna Synthesis, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/strand cdna synthesis/product/GE Healthcare
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    85
    GE Healthcare gst protein binding assay gst tagged rrp17 cdna
    Expression pattern of <t>RRP17.</t> (A) Northern blot analysis of RRP17 mRNA in adult mouse and human tissues. (B) Section of an E9.5 mouse embryo after in situ hybridization with an RRP17 <t>cDNA</t> labeled with dig-UTP. RRP17 transcripts are detected in the dorsal part of the neural tube (nt), skeletal myotome (m), and heart (h). (C) In situ hybridization of sections of E15.5 mouse brain. Left panels show in situ hybridization using RRP17 cDNA labeled with 35 S-UTP as probe and right panels show the same sections counterstained with hematoxylin. Bottom panels show magnification of cortex area (outlined by box in top panels). RRP17 transcript is detected in mature neurons. (n) neopalial cortex (future cerebral cortex), (i) intermediate zone; (v) ventricular zone; (lv) lateral ventricle; (p) pituitary. Bar: 500 μm for top panels, 100 μm for bottom panels. (D) In situ hybridization of sections of mouse heart and surrounding tissues at E13.5. Left panels show in situ hybridization using RRP17 cDNA labeled with 35 S-UTP as probe and right panels show the same section counterstained with hematoxylin. (h) heart; (d) dorsal root ganglia, and (s) gray matter of spinal cord. Bar: 500 μm for top panels, 100 μm for bottom panels.
    Gst Protein Binding Assay Gst Tagged Rrp17 Cdna, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gst protein binding assay gst tagged rrp17 cdna/product/GE Healthcare
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    92
    GE Healthcare cdna sequences
    Truncation and missense mutants of <t>DLC-1</t> for DLC-1 structure-function analyses. A schematic representation of the DLC-1 truncation constructs is shown. The full-length and various fragments of DLC-1 <t>cDNA</t> were subcloned into the EGFP-tagged expression vector, pEGFP-N1, resulting in the expression of carboxyl-terminal GFP-tagged fusion proteins.
    Cdna Sequences, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Characterization of androgen-responsive non-coding intronic RNAs . Selected intronic transcripts extended by RACE-PCR (A-D, red boxes) or gene-specific PCR (E, yellow box) are shown. Extended unspliced antisense intronic transcript fragments were mapped to the genomic sequence (A-E, gray lines) relative to the respective spliced protein-coding transcript expressed in the opposite strand (A-E, green arrows). Double-stranded cDNA clones spotted on the microarrays are represented as blue boxes (A-E). A previously described antisense unspliced intronic transcript mapping to GAS6 locus is shown (E, black arrow). Chromosome coordinates and the length of extended intronic transcript fragments are indicated. (F) Strand-specific multi-tissue northern blot using probes complementary to the antisense intronic transcript mapped in the GAS6 locus.

    Journal: BMC Biology

    Article Title: Androgen responsive intronic non-coding RNAs

    doi: 10.1186/1741-7007-5-4

    Figure Lengend Snippet: Characterization of androgen-responsive non-coding intronic RNAs . Selected intronic transcripts extended by RACE-PCR (A-D, red boxes) or gene-specific PCR (E, yellow box) are shown. Extended unspliced antisense intronic transcript fragments were mapped to the genomic sequence (A-E, gray lines) relative to the respective spliced protein-coding transcript expressed in the opposite strand (A-E, green arrows). Double-stranded cDNA clones spotted on the microarrays are represented as blue boxes (A-E). A previously described antisense unspliced intronic transcript mapping to GAS6 locus is shown (E, black arrow). Chromosome coordinates and the length of extended intronic transcript fragments are indicated. (F) Strand-specific multi-tissue northern blot using probes complementary to the antisense intronic transcript mapped in the GAS6 locus.

    Article Snippet: Microarray experiments The microarray platform used in this study was constructed using PCR-amplified cDNA fragments derived from ORESTES clones [ ], and spotted onto Type 7 STAR glass slides (GE Healthcare, Piscataway, NJ, USA) as previously described [ ].

    Techniques: Polymerase Chain Reaction, Sequencing, Clone Assay, Northern Blot

    Northern blot with total Cuscuta RNA . Cuscuta RNA from shoots (shoot) or shoot material with haustoria (haustoria). Upper panel: hybridization signal with a cuscutain-cDNA probe. Lower panel: Ethidium bromide stained total RNA to indicate even loading.

    Journal: BMC Plant Biology

    Article Title: Significance of Cuscutain, a cysteine protease from Cuscuta reflexa, in host-parasite interactions

    doi: 10.1186/1471-2229-10-227

    Figure Lengend Snippet: Northern blot with total Cuscuta RNA . Cuscuta RNA from shoots (shoot) or shoot material with haustoria (haustoria). Upper panel: hybridization signal with a cuscutain-cDNA probe. Lower panel: Ethidium bromide stained total RNA to indicate even loading.

    Article Snippet: 2 μg total RNA was employed for first strand cDNA synthesis (Ready-To-Go™ You-Prime First-Strand Beads; GE Healthcare) using 3'CDS-primer and SMARTIIA-oligo-primer from SMART™ cDNA synthesis system (Clontech). cDNA was PCR amplified applying the above mentioned primers and the product was cloned via TA cloning (TOPO TA Cloning® , Invitrogen).

    Techniques: Northern Blot, Hybridization, Staining

    Expression pattern of RRP17. (A) Northern blot analysis of RRP17 mRNA in adult mouse and human tissues. (B) Section of an E9.5 mouse embryo after in situ hybridization with an RRP17 cDNA labeled with dig-UTP. RRP17 transcripts are detected in the dorsal part of the neural tube (nt), skeletal myotome (m), and heart (h). (C) In situ hybridization of sections of E15.5 mouse brain. Left panels show in situ hybridization using RRP17 cDNA labeled with 35 S-UTP as probe and right panels show the same sections counterstained with hematoxylin. Bottom panels show magnification of cortex area (outlined by box in top panels). RRP17 transcript is detected in mature neurons. (n) neopalial cortex (future cerebral cortex), (i) intermediate zone; (v) ventricular zone; (lv) lateral ventricle; (p) pituitary. Bar: 500 μm for top panels, 100 μm for bottom panels. (D) In situ hybridization of sections of mouse heart and surrounding tissues at E13.5. Left panels show in situ hybridization using RRP17 cDNA labeled with 35 S-UTP as probe and right panels show the same section counterstained with hematoxylin. (h) heart; (d) dorsal root ganglia, and (s) gray matter of spinal cord. Bar: 500 μm for top panels, 100 μm for bottom panels.

    Journal: The Journal of Cell Biology

    Article Title: Regulation of atrial natriuretic peptide secretion by a novel Ras-like protein

    doi: 10.1083/jcb.200707101

    Figure Lengend Snippet: Expression pattern of RRP17. (A) Northern blot analysis of RRP17 mRNA in adult mouse and human tissues. (B) Section of an E9.5 mouse embryo after in situ hybridization with an RRP17 cDNA labeled with dig-UTP. RRP17 transcripts are detected in the dorsal part of the neural tube (nt), skeletal myotome (m), and heart (h). (C) In situ hybridization of sections of E15.5 mouse brain. Left panels show in situ hybridization using RRP17 cDNA labeled with 35 S-UTP as probe and right panels show the same sections counterstained with hematoxylin. Bottom panels show magnification of cortex area (outlined by box in top panels). RRP17 transcript is detected in mature neurons. (n) neopalial cortex (future cerebral cortex), (i) intermediate zone; (v) ventricular zone; (lv) lateral ventricle; (p) pituitary. Bar: 500 μm for top panels, 100 μm for bottom panels. (D) In situ hybridization of sections of mouse heart and surrounding tissues at E13.5. Left panels show in situ hybridization using RRP17 cDNA labeled with 35 S-UTP as probe and right panels show the same section counterstained with hematoxylin. (h) heart; (d) dorsal root ganglia, and (s) gray matter of spinal cord. Bar: 500 μm for top panels, 100 μm for bottom panels.

    Article Snippet: GST protein binding assay GST-tagged RRP17 cDNA was subcloned into pGEX-KG vector (GE Healthcare) and expressed in BL21(LysS) Escherichia coli grown in LB supplemented with 0.5 M sorbitol and 2.5 mM betaine.

    Techniques: Expressing, Northern Blot, In Situ Hybridization, Labeling

    Interaction of RRP17 with CAPS1. (A) Domains of CAPS1 protein. C2, calcium binding domain; PH, plextrin homology domain; MHD, munc homology domain or DUF1041; DCVD, dense-core vesicle binding domain. Bar indicates largest yeast-two hybrid clone found in screen. (B) GST pull-down assay consisted of 35 S-methionine-labeled CAPS1(670–1372) incubated with GST-RRP17, GST-RhoA or GST, with glutathione Sepharose beads. Bound CAPS1(670–1372) was detected using SDS-PAGE and autoradiography. (C) COS-7 cells were transfected with Flag-RRP17 and Flag-CAPS(670–1372) expression plasmids, lysed, separated into supernatant and pellet fractions. Equal aliquots of each fraction were separated on a 10% SDS-PAGE gel and immunoblotted with anti-FLAG antibody. (D) HeLa cells were transfected with plasmids encoding full-length CAPS1 and/or Flag-RRP17. Immunohistochemistry was performed using anti-CAPS (green) and anti-Flag (red) antibodies. Nuclei were counterstained with Hoechst 33238 (blue). Cells were visualized using confocal microscopy. Bar, 10 μm.

    Journal: The Journal of Cell Biology

    Article Title: Regulation of atrial natriuretic peptide secretion by a novel Ras-like protein

    doi: 10.1083/jcb.200707101

    Figure Lengend Snippet: Interaction of RRP17 with CAPS1. (A) Domains of CAPS1 protein. C2, calcium binding domain; PH, plextrin homology domain; MHD, munc homology domain or DUF1041; DCVD, dense-core vesicle binding domain. Bar indicates largest yeast-two hybrid clone found in screen. (B) GST pull-down assay consisted of 35 S-methionine-labeled CAPS1(670–1372) incubated with GST-RRP17, GST-RhoA or GST, with glutathione Sepharose beads. Bound CAPS1(670–1372) was detected using SDS-PAGE and autoradiography. (C) COS-7 cells were transfected with Flag-RRP17 and Flag-CAPS(670–1372) expression plasmids, lysed, separated into supernatant and pellet fractions. Equal aliquots of each fraction were separated on a 10% SDS-PAGE gel and immunoblotted with anti-FLAG antibody. (D) HeLa cells were transfected with plasmids encoding full-length CAPS1 and/or Flag-RRP17. Immunohistochemistry was performed using anti-CAPS (green) and anti-Flag (red) antibodies. Nuclei were counterstained with Hoechst 33238 (blue). Cells were visualized using confocal microscopy. Bar, 10 μm.

    Article Snippet: GST protein binding assay GST-tagged RRP17 cDNA was subcloned into pGEX-KG vector (GE Healthcare) and expressed in BL21(LysS) Escherichia coli grown in LB supplemented with 0.5 M sorbitol and 2.5 mM betaine.

    Techniques: Binding Assay, Pull Down Assay, Labeling, Incubation, SDS Page, Autoradiography, Transfection, Expressing, Immunohistochemistry, Confocal Microscopy

    Truncation and missense mutants of DLC-1 for DLC-1 structure-function analyses. A schematic representation of the DLC-1 truncation constructs is shown. The full-length and various fragments of DLC-1 cDNA were subcloned into the EGFP-tagged expression vector, pEGFP-N1, resulting in the expression of carboxyl-terminal GFP-tagged fusion proteins.

    Journal: The Journal of Biological Chemistry

    Article Title: Effects of Structure of Rho GTPase-activating Protein DLC-1 on Cell Morphology and Migration *Effects of Structure of Rho GTPase-activating Protein DLC-1 on Cell Morphology and Migration * S⃞

    doi: 10.1074/jbc.M800617200

    Figure Lengend Snippet: Truncation and missense mutants of DLC-1 for DLC-1 structure-function analyses. A schematic representation of the DLC-1 truncation constructs is shown. The full-length and various fragments of DLC-1 cDNA were subcloned into the EGFP-tagged expression vector, pEGFP-N1, resulting in the expression of carboxyl-terminal GFP-tagged fusion proteins.

    Article Snippet: For expression of glutathione S -transferase (GST) fusion recombinant protein, cDNA sequences for full-length DLC-1, DLC-1 ΔSAM, and DLC-1 RhoGAP domain were subcloned into the pGEX-5X-3 (GE Healthcare) bacterial expression vector.

    Techniques: Construct, Expressing, Plasmid Preparation