1d11 antibody Search Results


anti tgf β  (R&D Systems)
93
R&D Systems anti tgf β
Anti Tgf β, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1d11  (Novus Biologicals)
86
Novus Biologicals 1d11
Primary antibodies used
1d11, supplied by Novus Biologicals, 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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anti pkr antibody  (R&D Systems)
93
R&D Systems anti pkr antibody
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Anti Pkr Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 1 article reviews
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1d11 anti tgf β antibody  (R&D Systems)
90
R&D Systems 1d11 anti tgf β antibody
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
1d11 Anti Tgf β Antibody, supplied by R&D Systems, 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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antibodies against slit3  (R&D Systems)
94
R&D Systems antibodies against slit3
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Antibodies Against Slit3, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti transforming growth factor β tgf β alexa fluor 700  (R&D Systems)
93
R&D Systems anti transforming growth factor β tgf β alexa fluor 700
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Anti Transforming Growth Factor β Tgf β Alexa Fluor 700, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 1 article reviews
anti transforming growth factor β tgf β alexa fluor 700 - by Bioz Stars, 2026-03
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anti nkg2d antibody  (Novus Biologicals)
90
Novus Biologicals anti nkg2d antibody
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Anti Nkg2d Antibody, supplied by Novus Biologicals, 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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alexa fluor 488 conjugated anti tgf β1  (Novus Biologicals)
93
Novus Biologicals alexa fluor 488 conjugated anti tgf β1
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Alexa Fluor 488 Conjugated Anti Tgf β1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti tgf β 1 2 3 1d11  (R&D Systems)
95
R&D Systems anti tgf β 1 2 3 1d11
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Anti Tgf β 1 2 3 1d11, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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iii isoforms  (Novus Biologicals)
92
Novus Biologicals iii isoforms
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Iii Isoforms, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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il 4  (Novus Biologicals)
92
Novus Biologicals il 4
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Il 4, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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interleukin il 4  (Novus Biologicals)
91
Novus Biologicals interleukin il 4
(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous <t>PKR</t> and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts <t>were</t> <t>immunoprecipitated</t> at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.
Interleukin Il 4, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Primary antibodies used

Journal: Archivum Immunologiae et Therapiae Experimentalis

Article Title: Detection and Significance of Cytotoxic Cell Subsets in Biopsies of HCV-Infected Human Livers

doi: 10.1007/s00005-013-0258-6

Figure Lengend Snippet: Primary antibodies used

Article Snippet: NKG2D/CD314 , Mouse , Monoclonal , 1D11 , Novus Biologicals , NB100-65956.

Techniques:

(A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous PKR and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts were immunoprecipitated at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.

Journal: bioRxiv

Article Title: A frameshift mutation in the murine Prkra gene causes dystonia and exhibits abnormal cerebellar development and reduced eIF2α phosphorylation

doi: 10.1101/2024.06.04.597421

Figure Lengend Snippet: (A) dsRNA binding assay. dsRNA binding activity of WT PACT/RAX/RAX and lear-5J truncated protein was measured by a poly(I):poly(C)-agarose binding assay with in vitro translated 35 S-labeled proteins. T, total input; B, proteins bound to poly(I):poly(C)-agarose. Competition lanes 5 and 6: competition with 100-fold molar excess of single-stranded RNA (ss) or dsRNA (ds). The faint band below the parent PACT/RAX band represents products of in vitro translation from an internal methionine codon in lane 1. (B) Quantification of the dsRNA binding assay. Bands were quantified by phosphorimaging analysis, and % bound was calculated. Error bars: S.D. from three independent experiments. The p value (0.003) calculated using statistical analyses indicated significant difference between % dsRNA-binding of WT (blue bar) and lear-5J mutant (red bar) as indicated by the bracket. (C).Co-Immunoprecipitation of endogenous PKR and Flag-PACT/RAX or Flag-lear-5J overexpressed in HeLa cells. HeLa cells were transfected with Flag wt PACT/RAX or Flag-lear-5J in pCDNA3.1-expression constructs at 40% confluency and harvested 24-hour post transfection. Whole cell extracts were immunoprecipitated at 4°C overnight using 100 ng of anti-PKR antibody per IP. Samples were then analyzed via SDS-PAGE gel electrophoresis and western blot analysis probing for Flag tagged wt PACT/RAX or lear-5J (co-IP panel) using monoclonal anti-Flag-M2 (Sigma) antibody. To ascertain that an equal amount of protein was immunoprecipitated, blots were re-probed using an anti-PKR antibody (IP panel). Input blots without immunoprecipitation demonstrate equal amounts of each protein were present prior to IP.

Article Snippet: Whole cell extract was then immunoprecipitated overnight at 4°C on a rotating wheel using anti-PKR antibody (71/10, R&D Systems) conjugated to protein A sepharose beads (GE Healthcare) in IP buffer.

Techniques: Binding Assay, Activity Assay, In Vitro, Labeling, Mutagenesis, Immunoprecipitation, Transfection, Expressing, Construct, SDS Page, Nucleic Acid Electrophoresis, Western Blot, Co-Immunoprecipitation Assay

(A) Effect of lear-5J protein on PKR kinase activity. PKR kinase activity assay was performed using PKR immunoprecipitated from HeLa cells, recombinant lear-5J and wt PACT/RAX proteins, and 1 μCi of [γ- 32 P] ATP per reaction. Either pure recombinant lear-5J (left) or wt PACT/RAX (right) protein was added as activator in amounts indicated above each lane. Phosphorylated PKR was visualized after SDS-PAGE and phosphorimager analysis. (B) The truncated lear-5J protein inhibits PKR activation. PKR immunoprecipitated from HeLa cell extracts was activated with of polyI:polyC (lanes 2-5) or 4 ng of recombinant pure wt PACT/RAX protein (lanes 6-9). Increasing amounts of recombinant pure lear-5J protein (4 ng-400 ng) were added (lanes 2-9) as indicated to assess the effect on PKR activity. Lane 1: PKR activity without any added activator, lane 2: PKR activity in the presence of polyI:polyC, lanes 3-5: PKR activity in the presence of polyI:polyC and 4ng, 40 ng, or 400 ng of lear-5J protein, lane 6: PKR activity in the presence of 4ng of wt PACT/RAX protein, lanes 7-9: PKR activity in the presence of 4 ng of wt PACT/RAX and 4 ng, 40 ng or 400 ng of lear-5J protein. Phosphorylated proteins were analyzed by SDS-PAGE and phosphorimager analysis.

Journal: bioRxiv

Article Title: A frameshift mutation in the murine Prkra gene causes dystonia and exhibits abnormal cerebellar development and reduced eIF2α phosphorylation

doi: 10.1101/2024.06.04.597421

Figure Lengend Snippet: (A) Effect of lear-5J protein on PKR kinase activity. PKR kinase activity assay was performed using PKR immunoprecipitated from HeLa cells, recombinant lear-5J and wt PACT/RAX proteins, and 1 μCi of [γ- 32 P] ATP per reaction. Either pure recombinant lear-5J (left) or wt PACT/RAX (right) protein was added as activator in amounts indicated above each lane. Phosphorylated PKR was visualized after SDS-PAGE and phosphorimager analysis. (B) The truncated lear-5J protein inhibits PKR activation. PKR immunoprecipitated from HeLa cell extracts was activated with of polyI:polyC (lanes 2-5) or 4 ng of recombinant pure wt PACT/RAX protein (lanes 6-9). Increasing amounts of recombinant pure lear-5J protein (4 ng-400 ng) were added (lanes 2-9) as indicated to assess the effect on PKR activity. Lane 1: PKR activity without any added activator, lane 2: PKR activity in the presence of polyI:polyC, lanes 3-5: PKR activity in the presence of polyI:polyC and 4ng, 40 ng, or 400 ng of lear-5J protein, lane 6: PKR activity in the presence of 4ng of wt PACT/RAX protein, lanes 7-9: PKR activity in the presence of 4 ng of wt PACT/RAX and 4 ng, 40 ng or 400 ng of lear-5J protein. Phosphorylated proteins were analyzed by SDS-PAGE and phosphorimager analysis.

Article Snippet: Whole cell extract was then immunoprecipitated overnight at 4°C on a rotating wheel using anti-PKR antibody (71/10, R&D Systems) conjugated to protein A sepharose beads (GE Healthcare) in IP buffer.

Techniques: Activity Assay, Kinase Assay, Immunoprecipitation, Recombinant, SDS Page, Activation Assay