lentiviral constructs lentiviral constructs (New England Biolabs)


Name:
AgeI HF
Description:
AgeI HF 1 500 units
Catalog Number:
r3552l
Price:
298
Size:
1 500 units
Category:
Restriction Enzymes
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AgeI HF 1 500 units
https://www.bioz.com/result/lentiviral constructs lentiviral constructs/product/New England Biolabs
Average 91 stars, based on 3770 article reviews
Price from $9.99 to $1999.99
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1) Product Images from "Autoinflammatory mutation in NLRC4 reveals a leucine-rich repeat (LRR)–LRR oligomerization interface"
Article Title: Autoinflammatory mutation in NLRC4 reveals a leucine-rich repeat (LRR)–LRR oligomerization interface
Journal: The Journal of Allergy and Clinical Immunology
doi: 10.1016/j.jaci.2018.04.033
![... and LRR2). NLRC4 KO THP-1 cells transduced with lentiviral constructs expressing WT or mutant NLRC4 combined with ... Assessment of p.W655 NLRC4 in oligomerized form. A, Ribbon representation of NLRC4 in the oligomerized state (PDB Code 6B5B 24 ) with magnified inset areas highlighting 2 potential regions of interaction with p.W655 (LRR1 and LRR2). NLRC4 KO THP-1 cells transduced with lentiviral constructs expressing WT or mutant NLRC4 combined with LRR1 or LRR2 mutations. B-D, Twenty-four hours after transduction, cells were primed with Pam3CSK4 for 24 hours, and then propidium iodine (PI) staining was assessed by using flow cytometry to quantify cell death (Fig 7, B ), and IL-1β (Fig 7, C ) and IL-18 (Fig 7, D ) secretion were measured by ELISA. NLRC4 KO THP-1 cells were transduced with 1 mL of WT NLRC4 , LRR1 NLRC4 , or LRR2 NLRC4 virus or 2 mL of LRR1 NLRC4 virus (LRR1 [×2]). E-G, After 24 hours, cells were infected with a retroviral PrgI construct and cell death (Fig 7, E ), and IL-1β (Fig 7, F ) and IL-18 (Fig 7, G ) secretion assessed the following day. H and I , NLRC4 expression was assessed by using Western blot analysis. Data were pooled from at least 3 independent experiments. * P](https://storage.googleapis.com/bioz_article_images/PMC6281029/gr7.jpg)
Figure Legend Snippet: Assessment of p.W655 NLRC4 in oligomerized form. A, Ribbon representation of NLRC4 in the oligomerized state (PDB Code 6B5B 24 ) with magnified inset areas highlighting 2 potential regions of interaction with p.W655 (LRR1 and LRR2). NLRC4 KO THP-1 cells transduced with lentiviral constructs expressing WT or mutant NLRC4 combined with LRR1 or LRR2 mutations. B-D, Twenty-four hours after transduction, cells were primed with Pam3CSK4 for 24 hours, and then propidium iodine (PI) staining was assessed by using flow cytometry to quantify cell death (Fig 7, B ), and IL-1β (Fig 7, C ) and IL-18 (Fig 7, D ) secretion were measured by ELISA. NLRC4 KO THP-1 cells were transduced with 1 mL of WT NLRC4 , LRR1 NLRC4 , or LRR2 NLRC4 virus or 2 mL of LRR1 NLRC4 virus (LRR1 [×2]). E-G, After 24 hours, cells were infected with a retroviral PrgI construct and cell death (Fig 7, E ), and IL-1β (Fig 7, F ) and IL-18 (Fig 7, G ) secretion assessed the following day. H and I , NLRC4 expression was assessed by using Western blot analysis. Data were pooled from at least 3 independent experiments. * P
Techniques Used: Transduction, Construct, Expressing, Mutagenesis, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Infection, Western Blot
2) Product Images from "Generation of a New Frizzled 2 Flox Mouse Model to Clarify Its Role in Development"
Article Title: Generation of a New Frizzled 2 Flox Mouse Model to Clarify Its Role in Development
Journal: bioRxiv
doi: 10.1101/2021.01.27.428341

Figure Legend Snippet: Confirmation of Cre-mediated recombination of Fzd2 tm1Vari allele. A. Map of proposed Fzd2 tm1Vari recombination product. Xho1 and Age1 restriction sites (purple) were incorporated to aid in the validation of the allele. LoxP sites are indicated by red triangles, and 5’and 3’ untranslated regions (UTR) are shown. Primer binding sites for genotyping are shown. B . Limb micromass cultures from Fzd2 tmVari flox/flox were infected with Ad5CMV:eGFP (control) or Ad5CMV:Cre-eGFP at an MOI of 100. Brightfield and fluorescent images show infection efficiency. Ad5CMV:eGFP is cytoplasmic and Ad5CMV:Cre-eGFP is nuclear. Scale = 200um C . PCR-based genotyping for Cre or Fzd2 knockout (KO). Ad5CMV:Cre-eGFP infected cells amplified a product for Cre and Fzd2-KO. D . Single-cell suspensions were prepared from Ad:eGFP or Ad:Cre-eGFP infected micromass cultures, stained for Fzd2, and analyzed by flow cytometry. Cells were gated to discard debris and doublets. Live cells were determined by DAPI exclusion followed by gating on eGFP. FZD2 median fluorescent intensity (MFI) was compared between Ad:eGFP or Ad:Cre-eGFP infected micromass cultures. E . PCR-based allele-specific genotyping products for wild-type (292bp) and KO (485bp) products. The third lane includes a PCR product using primer set for KO only to be used for restriction digestions. F . Wild-type product was digested with XhoI and AgeI resulting in a wild-type uncut product (292bp). G . KO product was digested with XhoI and AgeI resulting in the expected digestion products: XhoI (196 and 291bp), AgeI (239 and 246bp). undig = undigested
Techniques Used: Binding Assay, Infection, Polymerase Chain Reaction, Knock-Out, Amplification, Staining, Flow Cytometry

Figure Legend Snippet: Generation and confirmation of Fzd2 tm1Vari allele. A. Map detailing the location of guide RNAs (indicated by the green and blue text on map and sequence) and regions of homology (ROH). B . Map of B6;C3H- Fzd2 tm1Vari intended modifications. LoxP sites were placed around the entire single exon Fzd2 gene. Xho1 and Age1 restriction sites (purple) were incorporated to aid in validation of the allele. LoxP sites are indicated by red triangles, and 5’ and 3’ untranslated regions (UTR) are shown. Primer binding sites for genotyping are shown in grey. C . PCR-based genotyping products for 5’ loxP site (top) and digestion with XhoI of flox product (bottom) results in expected products (193bp and 204 bp). D . PCR-based genotyping products for 3’ loxP site and digestion with AgeI of flox product results in the expected products (86bp and 238 bp). undig = undigested
Techniques Used: Sequencing, Binding Assay, Polymerase Chain Reaction
3) Product Images from "The RNA ligase RtcB reverses MazF-induced ribosome heterogeneity in Escherichia coli"
Article Title: The RNA ligase RtcB reverses MazF-induced ribosome heterogeneity in Escherichia coli
Journal: Nucleic Acids Research
doi: 10.1093/nar/gkw1018

Figure Legend Snippet: RtcB re-ligates RNA43 to purified 70S Δ43 ribosomes in vitro . ( A ) Schematic showing the in vitro ribosome repair assay (see text for details). ( B ) RNA43 AgeI used in the assay. Nucleotides changed in helix 45 and the binding site for primers Y12 (gray) and H17 (green) are indicated. ( C ) RT-PCR performed on RNA purified from 70S Δ43 ribosomes incubated in the absence of RtcB and RNA43 AgeI (lane 1), in the presence of either RNA43 AgeI (lane 2) or RtcB (lane 3), or both (lane 4) employing primer pairs specific for the central region of 16S rRNA (S7/X15; panel a), the 3΄end of 16S rRNA (S7/Y12; panel b), and specific for the AU nucleotide change present in 16S AgeI rRNA upon successful ligation (S7/H17; panel c). ( D ) AgeI restriction analysis and ( E ), sequencing of the RT-PCR product obtained with primer pair S7/H17 (shown in C, panel c, lane 4). Sequencing analyses from position 1509–1517 of the 16S rRNA of (a) rrsB , and (b) the RT-PCR product are shown. Nucleotides changed in RNA43 AgeI are underlined.
Techniques Used: Purification, In Vitro, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Incubation, Ligation, Sequencing
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