α cycb Search Results


anti-cycb  (Becton Dickinson)
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Becton Dickinson anti-cycb
Sequences in the <t>cycB</t> 3′ UTR are required for translational repression. (A) 3′ RACE PCR on the cycB transcript from RNA collected from wild-type testis, wild-type ovary, bam mutant testis (spermatogonia accumulate, and spermatocytes and spermatids are absent). Forward primer for 3′ RACE was nearly identical to primer #1 in C, just 4 bases longer. (B) RT-PCR from RNA collected from wild-type testis, wild-type ovary, bam mutant testis and rbp4 mutant testis, using primers #1-3, shown in C. Predicted products: 124 bp from short and long form; 227 bp from long form only. (C) Schematic of the cycB 3′ UTR as detected in ovary and spermatogonia versus in spermatocytes. NRE, Nanos response element. (D) A conserved proportion of the short cycB 3′ UTR, with mut9+5 variation created by site-directed mutagenesis (mutated nucleotides shown in lowercase). (E) Schematic of CycB-eYFP in vivo reporter. <t>(F-J)</t> <t>Anti-GFP</t> immunofluorescence on: (F) CycB-eYFP in wild-type testis, (G) CycB-eYFP in rbp4 mutant testis and (H) CycB-eYFP-mut9+5 in wild-type testis. White lines indicate early spermatocytes. Scale bar: 100 µm in H. (I,J) RT-PCR on CycB-eYFP, CycB-eYFP in rbp4 and CycB-eYFP-mut9+5 reporters. (I) Amplifying eYFP (313-bp expected product) and (control) GAPDH2 (100 bp) to assay reporter transcript levels. (J) Using an eYFP forward primer and cycB 3′ UTR reverse primers 2 and 3. Predicted products: 157 bp from short and long form; 260 bp from long form only – not detected. (K) Anti-GFP western blot of a biotin RNA pull-down from Rbp4-eYFP or Ubi-GFP testis extract. Wild-type and mutant biotin probes as indicated. (L) Quantification via ImageJ of three independent biotin RNA pull-downs from Rbp4-eYFP. The mean of the 10% input bands was set to 1; a value of 2 for the wild-type probe indicates that that probe pulled down ∼20% of the Rbp4-eYFP input. Error bars indicate s.e.m.
Anti Cycb, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Sequences in the cycB 3′ UTR are required for translational repression. (A) 3′ RACE PCR on the cycB transcript from RNA collected from wild-type testis, wild-type ovary, bam mutant testis (spermatogonia accumulate, and spermatocytes and spermatids are absent). Forward primer for 3′ RACE was nearly identical to primer #1 in C, just 4 bases longer. (B) RT-PCR from RNA collected from wild-type testis, wild-type ovary, bam mutant testis and rbp4 mutant testis, using primers #1-3, shown in C. Predicted products: 124 bp from short and long form; 227 bp from long form only. (C) Schematic of the cycB 3′ UTR as detected in ovary and spermatogonia versus in spermatocytes. NRE, Nanos response element. (D) A conserved proportion of the short cycB 3′ UTR, with mut9+5 variation created by site-directed mutagenesis (mutated nucleotides shown in lowercase). (E) Schematic of CycB-eYFP in vivo reporter. (F-J) Anti-GFP immunofluorescence on: (F) CycB-eYFP in wild-type testis, (G) CycB-eYFP in rbp4 mutant testis and (H) CycB-eYFP-mut9+5 in wild-type testis. White lines indicate early spermatocytes. Scale bar: 100 µm in H. (I,J) RT-PCR on CycB-eYFP, CycB-eYFP in rbp4 and CycB-eYFP-mut9+5 reporters. (I) Amplifying eYFP (313-bp expected product) and (control) GAPDH2 (100 bp) to assay reporter transcript levels. (J) Using an eYFP forward primer and cycB 3′ UTR reverse primers 2 and 3. Predicted products: 157 bp from short and long form; 260 bp from long form only – not detected. (K) Anti-GFP western blot of a biotin RNA pull-down from Rbp4-eYFP or Ubi-GFP testis extract. Wild-type and mutant biotin probes as indicated. (L) Quantification via ImageJ of three independent biotin RNA pull-downs from Rbp4-eYFP. The mean of the 10% input bands was set to 1; a value of 2 for the wild-type probe indicates that that probe pulled down ∼20% of the Rbp4-eYFP input. Error bars indicate s.e.m.

Journal: Development (Cambridge, England)

Article Title: Cell type-specific translational repression of Cyclin B during meiosis in males

doi: 10.1242/dev.122341

Figure Lengend Snippet: Sequences in the cycB 3′ UTR are required for translational repression. (A) 3′ RACE PCR on the cycB transcript from RNA collected from wild-type testis, wild-type ovary, bam mutant testis (spermatogonia accumulate, and spermatocytes and spermatids are absent). Forward primer for 3′ RACE was nearly identical to primer #1 in C, just 4 bases longer. (B) RT-PCR from RNA collected from wild-type testis, wild-type ovary, bam mutant testis and rbp4 mutant testis, using primers #1-3, shown in C. Predicted products: 124 bp from short and long form; 227 bp from long form only. (C) Schematic of the cycB 3′ UTR as detected in ovary and spermatogonia versus in spermatocytes. NRE, Nanos response element. (D) A conserved proportion of the short cycB 3′ UTR, with mut9+5 variation created by site-directed mutagenesis (mutated nucleotides shown in lowercase). (E) Schematic of CycB-eYFP in vivo reporter. (F-J) Anti-GFP immunofluorescence on: (F) CycB-eYFP in wild-type testis, (G) CycB-eYFP in rbp4 mutant testis and (H) CycB-eYFP-mut9+5 in wild-type testis. White lines indicate early spermatocytes. Scale bar: 100 µm in H. (I,J) RT-PCR on CycB-eYFP, CycB-eYFP in rbp4 and CycB-eYFP-mut9+5 reporters. (I) Amplifying eYFP (313-bp expected product) and (control) GAPDH2 (100 bp) to assay reporter transcript levels. (J) Using an eYFP forward primer and cycB 3′ UTR reverse primers 2 and 3. Predicted products: 157 bp from short and long form; 260 bp from long form only – not detected. (K) Anti-GFP western blot of a biotin RNA pull-down from Rbp4-eYFP or Ubi-GFP testis extract. Wild-type and mutant biotin probes as indicated. (L) Quantification via ImageJ of three independent biotin RNA pull-downs from Rbp4-eYFP. The mean of the 10% input bands was set to 1; a value of 2 for the wild-type probe indicates that that probe pulled down ∼20% of the Rbp4-eYFP input. Error bars indicate s.e.m.

Article Snippet: Antibody sources and dilutions: anti-CycB (1:30; F2F4, mouse, BD Biosciences), anti-HA (1:1000; 16B12, mouse, Covance), anti-GFP (1:3000; # {"type":"entrez-nucleotide","attrs":{"text":"A11122","term_id":"490966","term_text":"A11122"}} A11122 , rabbit, Invitrogen/Life Technologies), anti-PH3Thr3 (1:200; #07-424, rabbit, Millipore).

Techniques: Mutagenesis, Reverse Transcription Polymerase Chain Reaction, In Vivo, Immunofluorescence, Western Blot

A 31-amino acid domain in Rbp4 is required for binding to Fest. (A) Diagram of Rbp4 protein structure and truncated Rbp4 proteins tested in S2 cells for binding to Fest. RRM, RNA recognition motif. (B) Anti-HA, anti-Myc western blot of anti-Myc immunoprecipitations (top panels) and 10% input (bottom panels) from S2 cells transfected with HA-Fest alone or HA-Fest with various Myc-Rbp4 truncations, as indicated. (C) Alignment of Rbp4 residues 245-275 with Rbp4 homologs in other species. (D,E) Anti-GFP immunostaining of Rbp4-eYFP/+ testis (D) and Rbp4-eYFP/+; fest testis (E). Scale bar: 100 µm in E for D and E. (F) Biotin RNA pull-down from Rbp4-eYFP/+, fest testis, using the wild-type 130 nt cycB 3′ UTR probe.

Journal: Development (Cambridge, England)

Article Title: Cell type-specific translational repression of Cyclin B during meiosis in males

doi: 10.1242/dev.122341

Figure Lengend Snippet: A 31-amino acid domain in Rbp4 is required for binding to Fest. (A) Diagram of Rbp4 protein structure and truncated Rbp4 proteins tested in S2 cells for binding to Fest. RRM, RNA recognition motif. (B) Anti-HA, anti-Myc western blot of anti-Myc immunoprecipitations (top panels) and 10% input (bottom panels) from S2 cells transfected with HA-Fest alone or HA-Fest with various Myc-Rbp4 truncations, as indicated. (C) Alignment of Rbp4 residues 245-275 with Rbp4 homologs in other species. (D,E) Anti-GFP immunostaining of Rbp4-eYFP/+ testis (D) and Rbp4-eYFP/+; fest testis (E). Scale bar: 100 µm in E for D and E. (F) Biotin RNA pull-down from Rbp4-eYFP/+, fest testis, using the wild-type 130 nt cycB 3′ UTR probe.

Article Snippet: Antibody sources and dilutions: anti-CycB (1:30; F2F4, mouse, BD Biosciences), anti-HA (1:1000; 16B12, mouse, Covance), anti-GFP (1:3000; # {"type":"entrez-nucleotide","attrs":{"text":"A11122","term_id":"490966","term_text":"A11122"}} A11122 , rabbit, Invitrogen/Life Technologies), anti-PH3Thr3 (1:200; #07-424, rabbit, Millipore).

Techniques: Binding Assay, Western Blot, Transfection, Immunostaining