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binding buffer xbp  (Qiagen)


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    Structured Review

    Qiagen binding buffer xbp
    Binding Buffer Xbp, supplied by Qiagen, used in various techniques. Bioz Stars score: 94/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/binding+buffer/Buffer+XBP/us12624348-843-33-40
    Average 94 stars, based on 18 article reviews
    binding buffer xbp - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    Binding Assay:

    Article Title: Molecular detection of antibiotic-resistant Campylobacter spp. among broiler and layer chickens in Egypt: An outlook from a One Health concept.
    Article Snippet: Campylobacter is a zoonotic pathogen that poses a public health concern, with poultry acting as the main reservoir of human infection.. This study aimed to look into the prevalence and risk factors associated with antibiotic-resistant Campylobacter spp. found in chicken cloacal swabs using direct polymerase chain reaction (PCR) detection, highlighting their importance for public health.. In this study, 108 pooled chicken cloacal swabs from 270 broilers and 270 layers were included.

    Article Title: Ancient genomes from the siege and destruction of Middle Bronze Age Roca Vecchia (Apulia, Italy) shed light on Aegean contacts and conflicts
    Article Snippet: The purification steps were performed using the High Pure Viral Nucleic Acid Large Volume Kit columns (Roche). .. Ten times more Binding buffer (PB buffer (Qiagen)) than the sample volume was added and the concentrated samples were added to a 50 ml Roche column. ..

    Article Title: Mitotic chromosomes harbor cell type– and species-specific structural features within a universal loop array conformation
    Article Snippet: Instead of using lysis buffer to isolate nuclei, cells were pelleted by centrifugation for 5 min at 500 g at 4°C, resuspended in 50 μL of transposition reaction mix (25 μL of Tagmentation DNA buffer, 2.5 μL Tagment DNA enzyme [Illumina Tagment DNA TDE1 Enzyme and Buffer kits, Cat# 20034197], and 22.5 μL nuclease-free H 2 O), and incubated for 30 min at 37°C with gentle agitation. .. Reactions were stopped by adding the appropriate volume of Binding Buffer (Qiagen MinElute PCR kit), and the DNA was purified using the Qiagen MinElute PCR kit according to the manufacturer's protocol. ..

    Article Title: Ancient DNA from Palaeoloxodon naumanni in Japan reveals early evolution of Eurasian Palaeoloxodon
    Article Snippet: .. After centrifugation for 2 min at 2000 × g , the supernatant was incubated for 3 h at room temperature with 100 μL silica beads and 40 mL binding buffer (Qiagen PB buffer; 25 mM NaCl, 87 mM sodium acetate). .. After centrifugation for 5 min at 2000 × g , the supernatant was discarded, and the pellet was washed twice with 1 mL of 80% ethanol before eluting the DNA with 100 μL elution buffer (Qiagen).

    Article Title: Enhanced sedimentary ancient DNA (sedaDNA) retrieval from Antarctic diatoms
    Article Snippet: The supernatant (400–500 μl) was transferred to a new Lobind tube and subjected to DNA purification steps using solution C2 and C3 as per the manufacturer’s protocol (DNeasy Powersoil Kit, Qiagen). .. The hereby produced 500 μl of supernatant were then mixed with 500 μl EDTA supernatant and added to 4 ml of a modified binding buffer [Buffer QG (Qiagen) + Sodium Acetate (3 M) + Sodium Chloride (5 M) + Triton-x 100] with 100 μl silica solution (Brotherton et al., 2013). ..

    Article Title: Exploring the Evolution of the Cariogenic Oral Pathobiont Streptococcus mutans Using Ancient DNA
    Article Snippet: For the column-based extraction, the lysate was mixed with 10 mL binding buffer (5 M GuHCl, 40% Isoporpanol) and c. 400 μL 3M sodium acetate buffer (pH 5.2); following passage through a High Pure Extender Assembly (High Pure Viral Nucleic Acid Large Volume Kit, Roche), the DNA was treated with two ethanol washes and eluted in 100 μL TET (0.05% Tween-20 v/v) . .. For the in-solution extraction, the lysate was combined with a medium-sized silica suspension in 16 mL binding buffer (90% Qiagen QG buffer with c. 5 M guanidinium thiocyanate, 220 mM sodium acetate, 25 mM sodium chloride, 1% triton X100). ..

    Article Title: Ancient DNA from Palaeoloxodon naumanni in Japan reveals early evolution of Eurasian Palaeoloxodon .
    Article Snippet: .. After centrifugation for 2 min at 2000 × g, the supernatant was incubated for 3 h at room temperature with 100 μL silica beads and 40 mL binding buffer (Qiagen PB buffer; 25 mM NaCl, 87 mM sodium acetate). .. After centrifugation for 5 min at 2000 × g, the supernatant was discarded, and the pellet was washed twice with 1 mL of 80% ethanol before eluting the DNA with 100 μL elution buffer (Qiagen).

    Polymerase Chain Reaction:

    Article Title: Mitotic chromosomes harbor cell type– and species-specific structural features within a universal loop array conformation
    Article Snippet: Instead of using lysis buffer to isolate nuclei, cells were pelleted by centrifugation for 5 min at 500 g at 4°C, resuspended in 50 μL of transposition reaction mix (25 μL of Tagmentation DNA buffer, 2.5 μL Tagment DNA enzyme [Illumina Tagment DNA TDE1 Enzyme and Buffer kits, Cat# 20034197], and 22.5 μL nuclease-free H 2 O), and incubated for 30 min at 37°C with gentle agitation. .. Reactions were stopped by adding the appropriate volume of Binding Buffer (Qiagen MinElute PCR kit), and the DNA was purified using the Qiagen MinElute PCR kit according to the manufacturer's protocol. ..

    Purification:

    Article Title: Mitotic chromosomes harbor cell type– and species-specific structural features within a universal loop array conformation
    Article Snippet: Instead of using lysis buffer to isolate nuclei, cells were pelleted by centrifugation for 5 min at 500 g at 4°C, resuspended in 50 μL of transposition reaction mix (25 μL of Tagmentation DNA buffer, 2.5 μL Tagment DNA enzyme [Illumina Tagment DNA TDE1 Enzyme and Buffer kits, Cat# 20034197], and 22.5 μL nuclease-free H 2 O), and incubated for 30 min at 37°C with gentle agitation. .. Reactions were stopped by adding the appropriate volume of Binding Buffer (Qiagen MinElute PCR kit), and the DNA was purified using the Qiagen MinElute PCR kit according to the manufacturer's protocol. ..

    other:

    Article Title: FAM84B facilitates tau propagation via RYR3-mediated exocytosis in response to neuroinflammation
    Article Snippet: The membrane affinity-based exosome isolation process began with filtering the conditioned media through a 0.45 μm syringe filter (Pall Corporation, New York, NY, USA).

    Centrifugation:

    Article Title: Ancient DNA from Palaeoloxodon naumanni in Japan reveals early evolution of Eurasian Palaeoloxodon
    Article Snippet: .. After centrifugation for 2 min at 2000 × g , the supernatant was incubated for 3 h at room temperature with 100 μL silica beads and 40 mL binding buffer (Qiagen PB buffer; 25 mM NaCl, 87 mM sodium acetate). .. After centrifugation for 5 min at 2000 × g , the supernatant was discarded, and the pellet was washed twice with 1 mL of 80% ethanol before eluting the DNA with 100 μL elution buffer (Qiagen).

    Article Title: Ancient DNA from Palaeoloxodon naumanni in Japan reveals early evolution of Eurasian Palaeoloxodon .
    Article Snippet: .. After centrifugation for 2 min at 2000 × g, the supernatant was incubated for 3 h at room temperature with 100 μL silica beads and 40 mL binding buffer (Qiagen PB buffer; 25 mM NaCl, 87 mM sodium acetate). .. After centrifugation for 5 min at 2000 × g, the supernatant was discarded, and the pellet was washed twice with 1 mL of 80% ethanol before eluting the DNA with 100 μL elution buffer (Qiagen).

    Incubation:

    Article Title: Ancient DNA from Palaeoloxodon naumanni in Japan reveals early evolution of Eurasian Palaeoloxodon
    Article Snippet: .. After centrifugation for 2 min at 2000 × g , the supernatant was incubated for 3 h at room temperature with 100 μL silica beads and 40 mL binding buffer (Qiagen PB buffer; 25 mM NaCl, 87 mM sodium acetate). .. After centrifugation for 5 min at 2000 × g , the supernatant was discarded, and the pellet was washed twice with 1 mL of 80% ethanol before eluting the DNA with 100 μL elution buffer (Qiagen).

    Article Title: Ancient DNA from Palaeoloxodon naumanni in Japan reveals early evolution of Eurasian Palaeoloxodon .
    Article Snippet: .. After centrifugation for 2 min at 2000 × g, the supernatant was incubated for 3 h at room temperature with 100 μL silica beads and 40 mL binding buffer (Qiagen PB buffer; 25 mM NaCl, 87 mM sodium acetate). .. After centrifugation for 5 min at 2000 × g, the supernatant was discarded, and the pellet was washed twice with 1 mL of 80% ethanol before eluting the DNA with 100 μL elution buffer (Qiagen).

    Produced:

    Article Title: Enhanced sedimentary ancient DNA (sedaDNA) retrieval from Antarctic diatoms
    Article Snippet: The supernatant (400–500 μl) was transferred to a new Lobind tube and subjected to DNA purification steps using solution C2 and C3 as per the manufacturer’s protocol (DNeasy Powersoil Kit, Qiagen). .. The hereby produced 500 μl of supernatant were then mixed with 500 μl EDTA supernatant and added to 4 ml of a modified binding buffer [Buffer QG (Qiagen) + Sodium Acetate (3 M) + Sodium Chloride (5 M) + Triton-x 100] with 100 μl silica solution (Brotherton et al., 2013). ..

    Modification:

    Article Title: Enhanced sedimentary ancient DNA (sedaDNA) retrieval from Antarctic diatoms
    Article Snippet: The supernatant (400–500 μl) was transferred to a new Lobind tube and subjected to DNA purification steps using solution C2 and C3 as per the manufacturer’s protocol (DNeasy Powersoil Kit, Qiagen). .. The hereby produced 500 μl of supernatant were then mixed with 500 μl EDTA supernatant and added to 4 ml of a modified binding buffer [Buffer QG (Qiagen) + Sodium Acetate (3 M) + Sodium Chloride (5 M) + Triton-x 100] with 100 μl silica solution (Brotherton et al., 2013). ..

    Extraction:

    Article Title: Exploring the Evolution of the Cariogenic Oral Pathobiont Streptococcus mutans Using Ancient DNA
    Article Snippet: For the column-based extraction, the lysate was mixed with 10 mL binding buffer (5 M GuHCl, 40% Isoporpanol) and c. 400 μL 3M sodium acetate buffer (pH 5.2); following passage through a High Pure Extender Assembly (High Pure Viral Nucleic Acid Large Volume Kit, Roche), the DNA was treated with two ethanol washes and eluted in 100 μL TET (0.05% Tween-20 v/v) . .. For the in-solution extraction, the lysate was combined with a medium-sized silica suspension in 16 mL binding buffer (90% Qiagen QG buffer with c. 5 M guanidinium thiocyanate, 220 mM sodium acetate, 25 mM sodium chloride, 1% triton X100). ..

    Suspension:

    Article Title: Exploring the Evolution of the Cariogenic Oral Pathobiont Streptococcus mutans Using Ancient DNA
    Article Snippet: For the column-based extraction, the lysate was mixed with 10 mL binding buffer (5 M GuHCl, 40% Isoporpanol) and c. 400 μL 3M sodium acetate buffer (pH 5.2); following passage through a High Pure Extender Assembly (High Pure Viral Nucleic Acid Large Volume Kit, Roche), the DNA was treated with two ethanol washes and eluted in 100 μL TET (0.05% Tween-20 v/v) . .. For the in-solution extraction, the lysate was combined with a medium-sized silica suspension in 16 mL binding buffer (90% Qiagen QG buffer with c. 5 M guanidinium thiocyanate, 220 mM sodium acetate, 25 mM sodium chloride, 1% triton X100). ..



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    94
    Qiagen binding buffer xbp
    CRISPR/Cas9 induces DNA damage and apoptosis in PGCs. (A) Flow cytometry analysis 24 h after electroporation, quantifying the proportion of <t>Annexin</t> <t>V</t> + /PI + cells. The horizontal axis indicates PI and the vertical axis Annexin V. The upper-left quadrant (Annexin V + /PI + ) represents late apoptotic cells, and the lower-right quadrant (Annexin V + only) represents early apoptotic cells. Upper panels: results after electroporation with Cas9 + various gRNAs; lower panels: results with dCas9 + various gRNAs. (B) Bar graph of Annexin V + /PI + percentages across groups. Cas9 editing induced a highly significant increase in late apoptosis. (C) γ-H 2 AX foci (green) detected by immunofluorescence 24 h after electroporation. Foci appear as discrete nuclear puncta; nuclei are counterstained with DAPI (blue). Scale bar = 10 µm. (D) Quantification of γ-H 2 AX foci per cell. Cas9 targeting resulted in a significant increase in γ-H 2 AX foci per cell, whereas dCas9 with sgRNA did not. Statistical significance determined by one-way ANOVA (p-values are indicated in the figure).
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    ( A ) Immunoblotting of SIINFEKL peptide–pulsed Neuro2a cell lines cocultured with activated OT-I T cells for 4 hours at ratios of T cells to target cells indicated. ( B ) Schematic representation of fluorescence-activated cell sorting (FACS) based on Caspase-3(DEVD)NucView and annexin V (AnnV) staining, applies to (C) to (E). Created in BioRender; B. Liu (2026), https://BioRender.com/y3nlv0g . ( C ) Rate of outgrowth from cells recovered from FACS sort, calculated on the basis of number of initially plated cells. Data were compiled from five independent experiments. Paired t test. ( D and E ) Quantification of nucleotide fragments with greater than three, four, five, or six single-nucleotide variants (SNVs) from various recovered populations as indicated. ( F ) The design of the ATM–fluorescence resonance energy transfer (FRET) reporter. Created in BioRender; B. Liu (2026), https://BioRender.com/jqdhdx7 . ( G ) Quantification of FRET loss signal from Neuro2a expressing the ATM-FRET reporter shown in (F). Neuro2a cells were SIINFEKL peptide pulsed and then coincubated with OT-I T cells for 6 hours at 10:1 T cell–to–target cell ratio. Data were pooled from three independent experiments. Unpaired t test. ( H ) In vivo growth of Neuro2a cells expressing the ATM-FRET sensor. ATM-FRET loss populations were sorted post BH3m treatment (1 μM ABT737 and 1 μM S63845 ). Cells were sorted for FRET-loss population at 2.5 hours, and 1 × 10 4 cells were injected into NSG mice ( n = 5 animals per group). Kaplan-Meyer survival curves. WT-base and GE KO–base refer to ATM-FRET–expressing cells that were not treated with BH3m. Log-rank (Mantel-Cox) test. * P < 0.05 and **** P < 0.0001. C3, active caspase-3.

    Journal: Science Advances

    Article Title: Tumor gasdermin E acts as a rheostat to limit apoptotic cell survival and mutagenesis in anticancer immune environments

    doi: 10.1126/sciadv.aeb1137

    Figure Lengend Snippet: ( A ) Immunoblotting of SIINFEKL peptide–pulsed Neuro2a cell lines cocultured with activated OT-I T cells for 4 hours at ratios of T cells to target cells indicated. ( B ) Schematic representation of fluorescence-activated cell sorting (FACS) based on Caspase-3(DEVD)NucView and annexin V (AnnV) staining, applies to (C) to (E). Created in BioRender; B. Liu (2026), https://BioRender.com/y3nlv0g . ( C ) Rate of outgrowth from cells recovered from FACS sort, calculated on the basis of number of initially plated cells. Data were compiled from five independent experiments. Paired t test. ( D and E ) Quantification of nucleotide fragments with greater than three, four, five, or six single-nucleotide variants (SNVs) from various recovered populations as indicated. ( F ) The design of the ATM–fluorescence resonance energy transfer (FRET) reporter. Created in BioRender; B. Liu (2026), https://BioRender.com/jqdhdx7 . ( G ) Quantification of FRET loss signal from Neuro2a expressing the ATM-FRET reporter shown in (F). Neuro2a cells were SIINFEKL peptide pulsed and then coincubated with OT-I T cells for 6 hours at 10:1 T cell–to–target cell ratio. Data were pooled from three independent experiments. Unpaired t test. ( H ) In vivo growth of Neuro2a cells expressing the ATM-FRET sensor. ATM-FRET loss populations were sorted post BH3m treatment (1 μM ABT737 and 1 μM S63845 ). Cells were sorted for FRET-loss population at 2.5 hours, and 1 × 10 4 cells were injected into NSG mice ( n = 5 animals per group). Kaplan-Meyer survival curves. WT-base and GE KO–base refer to ATM-FRET–expressing cells that were not treated with BH3m. Log-rank (Mantel-Cox) test. * P < 0.05 and **** P < 0.0001. C3, active caspase-3.

    Article Snippet: Cells were spun down and resuspended in DMEM supplemented with annexin binding buffer (99902, Biotium) and stained with annexin V-647 and/or Caspase-3(DEVD)NucView-488, as indicated in the text, for 10 min at 37°C.

    Techniques: Western Blot, Fluorescence, FACS, Staining, Förster Resonance Energy Transfer, Expressing, In Vivo, Injection

    ( A ) Coculture (1:1) of parental B16F10 cells expressing pBabe-Cerulean (blue) and B16F10 cells expressing GsdmD-C3 and mCherry (red) (see movie S1). ( B ) Times of individual cell shrinkage and cell membrane lysis, as in (A). Unpaired t test. ( C ) Apoptotic phase (shrinkage-lysis interval) from (B). One-way ANOVA. ( D ) Quantification of annexin V + SIINFEKL peptide–pulsed B16F10 [as in (B) and (C)] that remains fluorescent when coincubated with activated OT-I T cells at 4 hours. Two-way ANOVA. ( E ) Quantification of SIINFEKL peptide–pulsed B16F10 cell populations with ectopic caspase-3-activatable GSDMD (GD-C3) by fluorescence signal at 24 hours of coincubation with activated OT-I T cells. ( F ) As in (E) with Gsdme. [(E) and (F)] Paired t test. ( G ) Number of GCamp6s spikes observed from SIINFEKL peptide–pulsed Neuro2a cells, coincubated with activated OT-I T cells, 10:1 T cell–to–target cell ratio. Images were taken every 2 min for a duration of 3 hours immediately following the additional of OT-I T cells, and numbers of observable GCamp6s spikes were counted and plotted (see movies S2 to S4 and fig. S5C). Unpaired t test. ( H ) Design for (I) and (J). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( I ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals ( n = 9 animals per group). Two-way ANOVA. ( J ) Kaplan-Meyer survival curves from C57BL/6 mice injected with Neuro2a cells ( n = 9 animals per group). ( K ) Design for (L). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( L ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals (control, n = 9 animals; Gsdme −/− , n = 8 animals). ( M ) Kaplan-Meyer survival curves from C57BL/6 mice rechallenged with Neuro2a cells [ n = 5 animals per group; log-rank (Mantel-Cox) test]. ( N ) As in (M) with B16F10 [ n = 12 animals in naïve group, 14 animals in rechallenge group; log-rank (Mantel-Cox) test]. * P < 0.05, ** P < 0.01, and **** P < 0.0001. iv, intravenous; ip, intraperitoneal.

    Journal: Science Advances

    Article Title: Tumor gasdermin E acts as a rheostat to limit apoptotic cell survival and mutagenesis in anticancer immune environments

    doi: 10.1126/sciadv.aeb1137

    Figure Lengend Snippet: ( A ) Coculture (1:1) of parental B16F10 cells expressing pBabe-Cerulean (blue) and B16F10 cells expressing GsdmD-C3 and mCherry (red) (see movie S1). ( B ) Times of individual cell shrinkage and cell membrane lysis, as in (A). Unpaired t test. ( C ) Apoptotic phase (shrinkage-lysis interval) from (B). One-way ANOVA. ( D ) Quantification of annexin V + SIINFEKL peptide–pulsed B16F10 [as in (B) and (C)] that remains fluorescent when coincubated with activated OT-I T cells at 4 hours. Two-way ANOVA. ( E ) Quantification of SIINFEKL peptide–pulsed B16F10 cell populations with ectopic caspase-3-activatable GSDMD (GD-C3) by fluorescence signal at 24 hours of coincubation with activated OT-I T cells. ( F ) As in (E) with Gsdme. [(E) and (F)] Paired t test. ( G ) Number of GCamp6s spikes observed from SIINFEKL peptide–pulsed Neuro2a cells, coincubated with activated OT-I T cells, 10:1 T cell–to–target cell ratio. Images were taken every 2 min for a duration of 3 hours immediately following the additional of OT-I T cells, and numbers of observable GCamp6s spikes were counted and plotted (see movies S2 to S4 and fig. S5C). Unpaired t test. ( H ) Design for (I) and (J). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( I ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals ( n = 9 animals per group). Two-way ANOVA. ( J ) Kaplan-Meyer survival curves from C57BL/6 mice injected with Neuro2a cells ( n = 9 animals per group). ( K ) Design for (L). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( L ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals (control, n = 9 animals; Gsdme −/− , n = 8 animals). ( M ) Kaplan-Meyer survival curves from C57BL/6 mice rechallenged with Neuro2a cells [ n = 5 animals per group; log-rank (Mantel-Cox) test]. ( N ) As in (M) with B16F10 [ n = 12 animals in naïve group, 14 animals in rechallenge group; log-rank (Mantel-Cox) test]. * P < 0.05, ** P < 0.01, and **** P < 0.0001. iv, intravenous; ip, intraperitoneal.

    Article Snippet: Cells were spun down and resuspended in DMEM supplemented with annexin binding buffer (99902, Biotium) and stained with annexin V-647 and/or Caspase-3(DEVD)NucView-488, as indicated in the text, for 10 min at 37°C.

    Techniques: Expressing, Membrane, Lysis, Fluorescence, Injection, Control

    CRISPR/Cas9 induces DNA damage and apoptosis in PGCs. (A) Flow cytometry analysis 24 h after electroporation, quantifying the proportion of Annexin V + /PI + cells. The horizontal axis indicates PI and the vertical axis Annexin V. The upper-left quadrant (Annexin V + /PI + ) represents late apoptotic cells, and the lower-right quadrant (Annexin V + only) represents early apoptotic cells. Upper panels: results after electroporation with Cas9 + various gRNAs; lower panels: results with dCas9 + various gRNAs. (B) Bar graph of Annexin V + /PI + percentages across groups. Cas9 editing induced a highly significant increase in late apoptosis. (C) γ-H 2 AX foci (green) detected by immunofluorescence 24 h after electroporation. Foci appear as discrete nuclear puncta; nuclei are counterstained with DAPI (blue). Scale bar = 10 µm. (D) Quantification of γ-H 2 AX foci per cell. Cas9 targeting resulted in a significant increase in γ-H 2 AX foci per cell, whereas dCas9 with sgRNA did not. Statistical significance determined by one-way ANOVA (p-values are indicated in the figure).

    Journal: Poultry Science

    Article Title: High genotoxicity of CRISPR/Cas9 versus limited efficacy of CRISPRi in chicken primordial germ cells

    doi: 10.1016/j.psj.2026.106722

    Figure Lengend Snippet: CRISPR/Cas9 induces DNA damage and apoptosis in PGCs. (A) Flow cytometry analysis 24 h after electroporation, quantifying the proportion of Annexin V + /PI + cells. The horizontal axis indicates PI and the vertical axis Annexin V. The upper-left quadrant (Annexin V + /PI + ) represents late apoptotic cells, and the lower-right quadrant (Annexin V + only) represents early apoptotic cells. Upper panels: results after electroporation with Cas9 + various gRNAs; lower panels: results with dCas9 + various gRNAs. (B) Bar graph of Annexin V + /PI + percentages across groups. Cas9 editing induced a highly significant increase in late apoptosis. (C) γ-H 2 AX foci (green) detected by immunofluorescence 24 h after electroporation. Foci appear as discrete nuclear puncta; nuclei are counterstained with DAPI (blue). Scale bar = 10 µm. (D) Quantification of γ-H 2 AX foci per cell. Cas9 targeting resulted in a significant increase in γ-H 2 AX foci per cell, whereas dCas9 with sgRNA did not. Statistical significance determined by one-way ANOVA (p-values are indicated in the figure).

    Article Snippet: After another centrifugation, the supernatant was removed and cells were resuspended in 500 μL of 1× Annexin V Binding Buffer (from Annexin V-FITC/PI Apoptosis Kit, Elabscience E-CK-A211).

    Techniques: CRISPR, Flow Cytometry, Electroporation, Immunofluorescence

    PGCs are more sensitive to DNA double-strand breaks than somatic cells. (A) Cell viability curves after 24 h treatment with increasing concentrations of etoposide (ETP; x-axis shows log 10 [ETP]). PGC viability dropped sharply even at low ETP doses, whereas CEF cells were more tolerant (viability data are mean ± SD of triplicates). (B) Apoptosis detection in PGCs treated with low-dose ETP. The proportion of Annexin V + /PI + cells (late apoptosis) was significantly elevated even at 0.03 µM ETP. (C) Western blot analysis of γ-H 2 AX protein in PGCs treated with varying ETP concentrations (left panel; β-actin as loading control). The bar graph (right) shows the ratio of γ-H 2 AX to β-actin band intensity, with a marked increase at 3 µM ETP (p-values are indicated in the figure). (D) Quantification of γ-H 2 AX foci per cell in THP-1 cells, male PGCs, and female PGCs after exposure to X-ray doses of 0, 2, 4, 6, and 8 Gy. After 48 h recovery, γ-H 2 AX foci increased significantly with higher radiation in THP-1, male PGCs, and female PGCs. Female PGC data points are red squares; male PGCs are blue squares; THP-1 are black circles. (E) Cell-cycle distribution of male vs. female PGCs after DNA damage. PGCs were irradiated (2, 4, 6 Gy), cultured 48 h, and analyzed by flow cytometry for cell-cycle phase (propidium iodide staining). Stacked bars show the percentage of cells in G 0 G 1 , S, and G 2 /M phases in untreated vs. irradiated cells. After damage, female PGCs predominantly arrested in G 2 /M (increased G 2 fraction), whereas male PGCs accumulated in S phase. Statistical significance by one-way ANOVA (p-values are indicated in the figure).

    Journal: Poultry Science

    Article Title: High genotoxicity of CRISPR/Cas9 versus limited efficacy of CRISPRi in chicken primordial germ cells

    doi: 10.1016/j.psj.2026.106722

    Figure Lengend Snippet: PGCs are more sensitive to DNA double-strand breaks than somatic cells. (A) Cell viability curves after 24 h treatment with increasing concentrations of etoposide (ETP; x-axis shows log 10 [ETP]). PGC viability dropped sharply even at low ETP doses, whereas CEF cells were more tolerant (viability data are mean ± SD of triplicates). (B) Apoptosis detection in PGCs treated with low-dose ETP. The proportion of Annexin V + /PI + cells (late apoptosis) was significantly elevated even at 0.03 µM ETP. (C) Western blot analysis of γ-H 2 AX protein in PGCs treated with varying ETP concentrations (left panel; β-actin as loading control). The bar graph (right) shows the ratio of γ-H 2 AX to β-actin band intensity, with a marked increase at 3 µM ETP (p-values are indicated in the figure). (D) Quantification of γ-H 2 AX foci per cell in THP-1 cells, male PGCs, and female PGCs after exposure to X-ray doses of 0, 2, 4, 6, and 8 Gy. After 48 h recovery, γ-H 2 AX foci increased significantly with higher radiation in THP-1, male PGCs, and female PGCs. Female PGC data points are red squares; male PGCs are blue squares; THP-1 are black circles. (E) Cell-cycle distribution of male vs. female PGCs after DNA damage. PGCs were irradiated (2, 4, 6 Gy), cultured 48 h, and analyzed by flow cytometry for cell-cycle phase (propidium iodide staining). Stacked bars show the percentage of cells in G 0 G 1 , S, and G 2 /M phases in untreated vs. irradiated cells. After damage, female PGCs predominantly arrested in G 2 /M (increased G 2 fraction), whereas male PGCs accumulated in S phase. Statistical significance by one-way ANOVA (p-values are indicated in the figure).

    Article Snippet: After another centrifugation, the supernatant was removed and cells were resuspended in 500 μL of 1× Annexin V Binding Buffer (from Annexin V-FITC/PI Apoptosis Kit, Elabscience E-CK-A211).

    Techniques: Western Blot, Control, Irradiation, Cell Culture, Flow Cytometry, Staining