transfex  (ATCC)


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    Structured Review

    ATCC transfex
    Lack of Cas9-mediated gene tagging of KIM-1 in RPTEC cells. RPTEC cells were transfected with jetPRIME ( A ), FuGENE-6 ( B ), and <t>TransfeX</t> ( C ) with TransfeX proving to be the most efficient. Despite good transfection efficiency, only 4 puromycin resistant clones were isolated. None of these clones demonstrated verified gene tagging via PCR ( D ). HK-2 clone 5 (HK-2-5) was used as a positive control. (-), untransfected cells.
    Transfex, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transfex/product/ATCC
    Average 95 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    transfex - by Bioz Stars, 2022-11
    95/100 stars

    Images

    1) Product Images from "CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line"

    Article Title: CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0204487

    Lack of Cas9-mediated gene tagging of KIM-1 in RPTEC cells. RPTEC cells were transfected with jetPRIME ( A ), FuGENE-6 ( B ), and TransfeX ( C ) with TransfeX proving to be the most efficient. Despite good transfection efficiency, only 4 puromycin resistant clones were isolated. None of these clones demonstrated verified gene tagging via PCR ( D ). HK-2 clone 5 (HK-2-5) was used as a positive control. (-), untransfected cells.
    Figure Legend Snippet: Lack of Cas9-mediated gene tagging of KIM-1 in RPTEC cells. RPTEC cells were transfected with jetPRIME ( A ), FuGENE-6 ( B ), and TransfeX ( C ) with TransfeX proving to be the most efficient. Despite good transfection efficiency, only 4 puromycin resistant clones were isolated. None of these clones demonstrated verified gene tagging via PCR ( D ). HK-2 clone 5 (HK-2-5) was used as a positive control. (-), untransfected cells.

    Techniques Used: Transfection, Clone Assay, Isolation, Polymerase Chain Reaction, Positive Control

    2) Product Images from "CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line"

    Article Title: CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0204487

    Lack of Cas9-mediated gene tagging of KIM-1 in RPTEC cells. RPTEC cells were transfected with jetPRIME ( A ), FuGENE-6 ( B ), and TransfeX ( C ) with TransfeX proving to be the most efficient. Despite good transfection efficiency, only 4 puromycin resistant clones were isolated. None of these clones demonstrated verified gene tagging via PCR ( D ). HK-2 clone 5 (HK-2-5) was used as a positive control. (-), untransfected cells.
    Figure Legend Snippet: Lack of Cas9-mediated gene tagging of KIM-1 in RPTEC cells. RPTEC cells were transfected with jetPRIME ( A ), FuGENE-6 ( B ), and TransfeX ( C ) with TransfeX proving to be the most efficient. Despite good transfection efficiency, only 4 puromycin resistant clones were isolated. None of these clones demonstrated verified gene tagging via PCR ( D ). HK-2 clone 5 (HK-2-5) was used as a positive control. (-), untransfected cells.

    Techniques Used: Transfection, Clone Assay, Isolation, Polymerase Chain Reaction, Positive Control

    3) Product Images from "CIRCADIAN RHYTHMICITY: A FUNCTIONAL CONNECTION BETWEEN DIFFERENTIATED EMBRYONIC CHONDROCYTE-1 (DEC1) AND SMALL HETERODIMER PARTNER (SHP)"

    Article Title: CIRCADIAN RHYTHMICITY: A FUNCTIONAL CONNECTION BETWEEN DIFFERENTIATED EMBRYONIC CHONDROCYTE-1 (DEC1) AND SMALL HETERODIMER PARTNER (SHP)

    Journal: Archives of biochemistry and biophysics

    doi: 10.1016/j.abb.2017.08.004

    Inverse expression between DEC1 and SHP (A) Suppressed expression of SHP by DEC1 HepG2 cells were seeded in 8-well chamber slides (2×10 4 cells/ well) and cultured for 24 h. Cells were then transfected with Flag-DEC1 (500 ng) or the vector by TransfeX reagent. The transfected cells were cultured for another 24 h and subsequently fixed with 4% paraformaldehyde for 10 min at pH 7.4. Cells were permeabilized with 0.1% Triton X-100 for 10 min. Chamber slides were incubated with 1% BSA (2mg/mL) for 1 h then incubated overnight with anti-SHP antibody or anti-Flag antibody. The anti-SHP antibody was located by Alexa Fluor ® 488 conjugated goat anti-rabbit IgG (green), whereas the anti-Flag antibody with Alexa Fluor ® 555 conjugated goat anti-mouse IgG (red). The slides were then mounted with ProLong Gold Antifade Mountant with DAPI (Blue). Cells were then imaged with confocal microscope. To provide more quantitative information, same experiments were performed in 6-well plates and cell lysates were prepared. Lysates (10 µg) were analyzed by Western blotting for the expression of SHP and DEC1 (Right panel). (B) Regulated expression of DEC1 and SHP by valproate in serum-shocked circadian induction HepG2 cells were seeded in 6- or 24-well plates at a density of 6×10 5 or 1.5×10 5 cells/well and cultured to reach 75% confluency. Cells were then shocked with media containing 50% horse serum for 2 h. Thereafter, the shocked cells were cultured in normal media or the same media containing valproate at 2 mM. Cells were harvested at a 6 h interval. The expression of DEC1 and SHP was determined by RT-qPCR with Taqman probes.
    Figure Legend Snippet: Inverse expression between DEC1 and SHP (A) Suppressed expression of SHP by DEC1 HepG2 cells were seeded in 8-well chamber slides (2×10 4 cells/ well) and cultured for 24 h. Cells were then transfected with Flag-DEC1 (500 ng) or the vector by TransfeX reagent. The transfected cells were cultured for another 24 h and subsequently fixed with 4% paraformaldehyde for 10 min at pH 7.4. Cells were permeabilized with 0.1% Triton X-100 for 10 min. Chamber slides were incubated with 1% BSA (2mg/mL) for 1 h then incubated overnight with anti-SHP antibody or anti-Flag antibody. The anti-SHP antibody was located by Alexa Fluor ® 488 conjugated goat anti-rabbit IgG (green), whereas the anti-Flag antibody with Alexa Fluor ® 555 conjugated goat anti-mouse IgG (red). The slides were then mounted with ProLong Gold Antifade Mountant with DAPI (Blue). Cells were then imaged with confocal microscope. To provide more quantitative information, same experiments were performed in 6-well plates and cell lysates were prepared. Lysates (10 µg) were analyzed by Western blotting for the expression of SHP and DEC1 (Right panel). (B) Regulated expression of DEC1 and SHP by valproate in serum-shocked circadian induction HepG2 cells were seeded in 6- or 24-well plates at a density of 6×10 5 or 1.5×10 5 cells/well and cultured to reach 75% confluency. Cells were then shocked with media containing 50% horse serum for 2 h. Thereafter, the shocked cells were cultured in normal media or the same media containing valproate at 2 mM. Cells were harvested at a 6 h interval. The expression of DEC1 and SHP was determined by RT-qPCR with Taqman probes.

    Techniques Used: Expressing, Cell Culture, Transfection, Plasmid Preparation, Incubation, Microscopy, Western Blot, Quantitative RT-PCR

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    ATCC transfex transfection reagent
    A universal strategy for simple and rapid genomic editing of cell populations. ( A ) Drosophila ovarian somatic sheath cells (OSC) represent a unique but delicate model to study Piwi–piRNA mechanism ex vivo. OSC represent somatic follicle cells (FC) of the Drosophila ovary, and express one of the three Drosophila PIWI proteins, Piwi. PiRNAs are generated from long piRNA cluster transcripts by the endonuclease Zucchini (Zuc). Mature Piwi–piRNA silencing complexes transition into the nucleus, recognize nascent transposon transcripts by base-pairing complementarity and induce epigenetic silencing. ( B ) Endogenous tagging of piwi in OSC. An sgRNA was designed to target the endogenous piwi gene in the vicinity of the start codon (ATG). The donor construct for homologous repair contained a FLAG-HA (FH)-tag and a puromycin resistance gene. The FH-tag was fused in frame with piwi's open reading frame (ORF) to generate an endogenously N-terminally tagged protein (eFH-). The puromycin resistance was placed into a synthetic intron and transcribed from the opposite genomic strand. The edited allele is designed to express two independent transcripts: The piwi transcript remains under the control of the endogenous promoter and contains an additional intron and a tag. The mature modified mRNA differs from the wt piwi mRNA only by an additional exon-exon junction and the Flag-HA tag. The second transcript is independently generated from the opposite genomic strand and produces an mRNA encoding a puromycin resistance under the control of an Actin promoter. ( C ) Rapid and simple generation of stably edited OSC:eFH-piwi. Ovarian somatic sheath cells (OSC) were transfected with an expression plasmid for the sgRNA, the Cas9 endonuclease, and the donor plasmid. Cells were treated with SCR7, an inhibitor of non-homologous end joining (NHEJ) to increase the probability for homologous repair. Antibiotic selection with Puromycin (Puro) was started 48 h after <t>transfection.</t> After 2–3 weeks, a puromycin resistant cell population has repopulated the dish
    Transfex Transfection Reagent, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transfex transfection reagent/product/ATCC
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transfex transfection reagent - by Bioz Stars, 2022-11
    94/100 stars
      Buy from Supplier

    94
    ATCC transfex transfection kit
    A universal strategy for simple and rapid genomic editing of cell populations. ( A ) Drosophila ovarian somatic sheath cells (OSC) represent a unique but delicate model to study Piwi–piRNA mechanism ex vivo. OSC represent somatic follicle cells (FC) of the Drosophila ovary, and express one of the three Drosophila PIWI proteins, Piwi. PiRNAs are generated from long piRNA cluster transcripts by the endonuclease Zucchini (Zuc). Mature Piwi–piRNA silencing complexes transition into the nucleus, recognize nascent transposon transcripts by base-pairing complementarity and induce epigenetic silencing. ( B ) Endogenous tagging of piwi in OSC. An sgRNA was designed to target the endogenous piwi gene in the vicinity of the start codon (ATG). The donor construct for homologous repair contained a FLAG-HA (FH)-tag and a puromycin resistance gene. The FH-tag was fused in frame with piwi's open reading frame (ORF) to generate an endogenously N-terminally tagged protein (eFH-). The puromycin resistance was placed into a synthetic intron and transcribed from the opposite genomic strand. The edited allele is designed to express two independent transcripts: The piwi transcript remains under the control of the endogenous promoter and contains an additional intron and a tag. The mature modified mRNA differs from the wt piwi mRNA only by an additional exon-exon junction and the Flag-HA tag. The second transcript is independently generated from the opposite genomic strand and produces an mRNA encoding a puromycin resistance under the control of an Actin promoter. ( C ) Rapid and simple generation of stably edited OSC:eFH-piwi. Ovarian somatic sheath cells (OSC) were transfected with an expression plasmid for the sgRNA, the Cas9 endonuclease, and the donor plasmid. Cells were treated with SCR7, an inhibitor of non-homologous end joining (NHEJ) to increase the probability for homologous repair. Antibiotic selection with Puromycin (Puro) was started 48 h after <t>transfection.</t> After 2–3 weeks, a puromycin resistant cell population has repopulated the dish
    Transfex Transfection Kit, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transfex transfection kit/product/ATCC
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    transfex transfection kit - by Bioz Stars, 2022-11
    94/100 stars
      Buy from Supplier

    Image Search Results


    A universal strategy for simple and rapid genomic editing of cell populations. ( A ) Drosophila ovarian somatic sheath cells (OSC) represent a unique but delicate model to study Piwi–piRNA mechanism ex vivo. OSC represent somatic follicle cells (FC) of the Drosophila ovary, and express one of the three Drosophila PIWI proteins, Piwi. PiRNAs are generated from long piRNA cluster transcripts by the endonuclease Zucchini (Zuc). Mature Piwi–piRNA silencing complexes transition into the nucleus, recognize nascent transposon transcripts by base-pairing complementarity and induce epigenetic silencing. ( B ) Endogenous tagging of piwi in OSC. An sgRNA was designed to target the endogenous piwi gene in the vicinity of the start codon (ATG). The donor construct for homologous repair contained a FLAG-HA (FH)-tag and a puromycin resistance gene. The FH-tag was fused in frame with piwi's open reading frame (ORF) to generate an endogenously N-terminally tagged protein (eFH-). The puromycin resistance was placed into a synthetic intron and transcribed from the opposite genomic strand. The edited allele is designed to express two independent transcripts: The piwi transcript remains under the control of the endogenous promoter and contains an additional intron and a tag. The mature modified mRNA differs from the wt piwi mRNA only by an additional exon-exon junction and the Flag-HA tag. The second transcript is independently generated from the opposite genomic strand and produces an mRNA encoding a puromycin resistance under the control of an Actin promoter. ( C ) Rapid and simple generation of stably edited OSC:eFH-piwi. Ovarian somatic sheath cells (OSC) were transfected with an expression plasmid for the sgRNA, the Cas9 endonuclease, and the donor plasmid. Cells were treated with SCR7, an inhibitor of non-homologous end joining (NHEJ) to increase the probability for homologous repair. Antibiotic selection with Puromycin (Puro) was started 48 h after transfection. After 2–3 weeks, a puromycin resistant cell population has repopulated the dish

    Journal: Nucleic Acids Research

    Article Title: Functional editing of endogenous genes through rapid selection of cell pools (Rapid generation of endogenously tagged genes in Drosophila ovarian somatic sheath cells)

    doi: 10.1093/nar/gkac448

    Figure Lengend Snippet: A universal strategy for simple and rapid genomic editing of cell populations. ( A ) Drosophila ovarian somatic sheath cells (OSC) represent a unique but delicate model to study Piwi–piRNA mechanism ex vivo. OSC represent somatic follicle cells (FC) of the Drosophila ovary, and express one of the three Drosophila PIWI proteins, Piwi. PiRNAs are generated from long piRNA cluster transcripts by the endonuclease Zucchini (Zuc). Mature Piwi–piRNA silencing complexes transition into the nucleus, recognize nascent transposon transcripts by base-pairing complementarity and induce epigenetic silencing. ( B ) Endogenous tagging of piwi in OSC. An sgRNA was designed to target the endogenous piwi gene in the vicinity of the start codon (ATG). The donor construct for homologous repair contained a FLAG-HA (FH)-tag and a puromycin resistance gene. The FH-tag was fused in frame with piwi's open reading frame (ORF) to generate an endogenously N-terminally tagged protein (eFH-). The puromycin resistance was placed into a synthetic intron and transcribed from the opposite genomic strand. The edited allele is designed to express two independent transcripts: The piwi transcript remains under the control of the endogenous promoter and contains an additional intron and a tag. The mature modified mRNA differs from the wt piwi mRNA only by an additional exon-exon junction and the Flag-HA tag. The second transcript is independently generated from the opposite genomic strand and produces an mRNA encoding a puromycin resistance under the control of an Actin promoter. ( C ) Rapid and simple generation of stably edited OSC:eFH-piwi. Ovarian somatic sheath cells (OSC) were transfected with an expression plasmid for the sgRNA, the Cas9 endonuclease, and the donor plasmid. Cells were treated with SCR7, an inhibitor of non-homologous end joining (NHEJ) to increase the probability for homologous repair. Antibiotic selection with Puromycin (Puro) was started 48 h after transfection. After 2–3 weeks, a puromycin resistant cell population has repopulated the dish

    Article Snippet: Transfection was done using the TransfeX transfection reagent (ATCC, ACS-4005).

    Techniques: Ex Vivo, Generated, Construct, Modification, Stable Transfection, Transfection, Expressing, Plasmid Preparation, Non-Homologous End Joining, Selection