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duolink blocking solution (1% bsa)  (Millipore)


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    Millipore duolink blocking solution (1% bsa)
    Duolink Blocking Solution (1% Bsa), supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/duolink+blocking+solution/duolink+blocking+solution/bio_rxiv__2025__07__15__664851-303-10-15
    Average 90 stars, based on 1 article reviews
    duolink blocking solution (1% bsa) - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: KIBRA anchoring the action of PKMζ maintains the persistence of memory
    Article Snippet: Slices were then washed with PBS (pH 7.4) and cut into 20 μM sections using a Leica VT 1200S vibratome. .. Free-floating sections were permeabilized in 96-well plates with PBS-TX100 for 6X 10 min at 20°C and blocked with Duolink Blocking Solution (Sigma-Aldrich, DUO82007-8 ml) for 1 hour at 37°C in a preheated humidity chamber. ..

    Article Title: Nonreceptor tyrosine kinase ABL1 regulates lysosomal acidification by phosphorylating the ATP6V1B2 subunit of the vacuolar-type H + -ATPase.
    Article Snippet: The Duolink in situ proximity ligation assay (Sigma-Aldrich, DUO92008) enables the endogenous detection of protein interactions and posttranslational modi cations of proteins at the single-molecule level in xed cell and tissue samples. .. Brie y, A549 cells plated on glass coverslips were xed with 4% formaldehyde, and the cells were permeabilized with 0.2% Triton X-100 in PBS for 15 min. After blocking with Duolink Blocking Solution(Sigma-Aldrich, DUO82007), the cells were incubated with rabbit antiATP6V1B2 (Sigma-Aldrich, HPA00814, 1:100 dilution) and mouse anti-ABL1 (8E9, Santa Cruz Biotechnology, sc -56,887, 1:50 dilution) or mouse anti-phosphotyrosine (4G10, Millipore, 05–321, 1:100 dilution) or mouse anti-ATP6V0D1 (Abcam, ab56441, 1:100 dilution) primary antibodies and then incubated with oligonucleotide-labeled anti-mouse IgG (Sigma-Aldrich, DUO92004) and anti-rabbit IgG (Sigma-Aldrich, DUO92002) according to the manufacturer’s instructions for PLA. ..

    Article Title: PARP10 promotes the repair of nascent strand DNA gaps through RAD18 mediated translesion synthesis.
    Article Snippet: .. Cells were then permeabilized with 0.5% Triton for 10min at 4 °C, washed with PBS, fixed at room temperature with 3% paraformaldehyde in PBS for 10min, washed again in PBS and then blocked in Duolink blocking solution (Millipore Sigma DUO82007) for 1 hr at 37 °C, and incubated overnight at 4 °C with primary antibodies. .. Antibodies used were: PARP10 (Abcam ab70800); MAR AbD33204 (BioRad HCA354) and RAD18 (Cell Signaling Technology 9040).

    Article Title: PARP10 promotes the repair of nascent strand DNA gaps through RAD18 mediated translesion synthesis
    Article Snippet: .. Cells were then permeabilized with 0.5% Triton for 10 min at 4 °C, washed with PBS, fixed at room temperature with 3% paraformaldehyde in PBS for 10 min, washed again in PBS and then blocked in Duolink blocking solution (Millipore Sigma DUO82007) for 1 hr at 37 °C, and incubated overnight at 4 °C with primary antibodies. .. Antibodies used were: PARP10 (Abcam ab70800); MAR AbD33204 (BioRad HCA354) and RAD18 (Cell Signaling Technology 9040).

    Article Title: Remodeling of anti-tumor immunity with antibodies targeting a p53 mutant
    Article Snippet: .. Following three PBS washes, samples were blocked with Duolink Blocking Solution (Sigma-Aldrich, DUO82007) for 1 h at 37 °C. .. Primary antibodies, including the purified E285K-mAb and TRIM21-mAb (Abcam, ab207728), were added at 5 μg/mL in antibody dilution solution at 37 °C for 1 h. After washing twice with PBS, samples were incubated with the Duolink In Situ PLA Probe Anti-Mouse MINUS (Sigma-Aldrich, DUO92001) and Duolink In Situ PLA Probe Anti-Rabbit PLUS (Sigma-Aldrich, DUO92005) in PLA antibody diluent (MilliporeSigma, DUO82008) for 1 h at 37 °C.

    Article Title: Genome-Wide Mapping of RNA-Protein Associations via Sequencing
    Article Snippet: The cells were washed for 10 min twice with 2X SSC (Invitrogen™ 15557044) and twice with 1x PBS pH 7.4 (Gibco™, 70011044) at room temperature. .. Antibodies were blocked by incubation with 100 μL of Duolink Blocking Solution (Sigma-Aldrich, DUO92101-1KT) according to the manufacturer’s recommendations. .. Incubation with antibodies: Two oligo-conjugated antibodies, including the antibody of protein of interest, which is conjugated with Oligo A from DUOLINK red kit (Sigma-Aldrich, DUO92008-100RXN), and anti-Digoxin, which is conjugated with Oligo B from DUOLINK red kit, (Sigma-Aldrich, DUO92008-100RXN), were mixed in Probemaker PLA Probe Diluent (Sigma-Aldrich, DUO82036) to a total volume of 200 μL and added to the fixed cells.

    Article Title: ESR1 and p53 interactome alteration defines mechanisms of tamoxifen response in luminal breast cancer
    Article Snippet: .. After hydration, TMAs were washed twice with tris-buffered saline with Tween 20 (TBST) followed by antigen retrieval using antigen retrieval buffer (EnVision FLEX Target Retrieval Solution, High pH (50×) (Dako Omnis) GV804) in steamer at 100°C for 1 h. TMA slides were cooled at room temperature and further quenched by Duolink hydrogen peroxide (Sigma-DUO82054) for 10 min followed by blocking by Duolink blocking solution (Sigma-DUO82007) for 5 min. Primary antibodies of ERα (HC20-1:100) and p53 (DO1-1:100) were diluted appropriately in antibody dilution buffer applied on each TMA (50 μL) and incubated further for 1 h in a humidified chamber. ..

    Incubation:

    Article Title: Nonreceptor tyrosine kinase ABL1 regulates lysosomal acidification by phosphorylating the ATP6V1B2 subunit of the vacuolar-type H + -ATPase.
    Article Snippet: The Duolink in situ proximity ligation assay (Sigma-Aldrich, DUO92008) enables the endogenous detection of protein interactions and posttranslational modi cations of proteins at the single-molecule level in xed cell and tissue samples. .. Brie y, A549 cells plated on glass coverslips were xed with 4% formaldehyde, and the cells were permeabilized with 0.2% Triton X-100 in PBS for 15 min. After blocking with Duolink Blocking Solution(Sigma-Aldrich, DUO82007), the cells were incubated with rabbit antiATP6V1B2 (Sigma-Aldrich, HPA00814, 1:100 dilution) and mouse anti-ABL1 (8E9, Santa Cruz Biotechnology, sc -56,887, 1:50 dilution) or mouse anti-phosphotyrosine (4G10, Millipore, 05–321, 1:100 dilution) or mouse anti-ATP6V0D1 (Abcam, ab56441, 1:100 dilution) primary antibodies and then incubated with oligonucleotide-labeled anti-mouse IgG (Sigma-Aldrich, DUO92004) and anti-rabbit IgG (Sigma-Aldrich, DUO92002) according to the manufacturer’s instructions for PLA. ..

    Article Title: PARP10 promotes the repair of nascent strand DNA gaps through RAD18 mediated translesion synthesis.
    Article Snippet: .. Cells were then permeabilized with 0.5% Triton for 10min at 4 °C, washed with PBS, fixed at room temperature with 3% paraformaldehyde in PBS for 10min, washed again in PBS and then blocked in Duolink blocking solution (Millipore Sigma DUO82007) for 1 hr at 37 °C, and incubated overnight at 4 °C with primary antibodies. .. Antibodies used were: PARP10 (Abcam ab70800); MAR AbD33204 (BioRad HCA354) and RAD18 (Cell Signaling Technology 9040).

    Article Title: PARP10 promotes the repair of nascent strand DNA gaps through RAD18 mediated translesion synthesis
    Article Snippet: .. Cells were then permeabilized with 0.5% Triton for 10 min at 4 °C, washed with PBS, fixed at room temperature with 3% paraformaldehyde in PBS for 10 min, washed again in PBS and then blocked in Duolink blocking solution (Millipore Sigma DUO82007) for 1 hr at 37 °C, and incubated overnight at 4 °C with primary antibodies. .. Antibodies used were: PARP10 (Abcam ab70800); MAR AbD33204 (BioRad HCA354) and RAD18 (Cell Signaling Technology 9040).

    Article Title: Genome-Wide Mapping of RNA-Protein Associations via Sequencing
    Article Snippet: The cells were washed for 10 min twice with 2X SSC (Invitrogen™ 15557044) and twice with 1x PBS pH 7.4 (Gibco™, 70011044) at room temperature. .. Antibodies were blocked by incubation with 100 μL of Duolink Blocking Solution (Sigma-Aldrich, DUO92101-1KT) according to the manufacturer’s recommendations. .. Incubation with antibodies: Two oligo-conjugated antibodies, including the antibody of protein of interest, which is conjugated with Oligo A from DUOLINK red kit (Sigma-Aldrich, DUO92008-100RXN), and anti-Digoxin, which is conjugated with Oligo B from DUOLINK red kit, (Sigma-Aldrich, DUO92008-100RXN), were mixed in Probemaker PLA Probe Diluent (Sigma-Aldrich, DUO82036) to a total volume of 200 μL and added to the fixed cells.

    Article Title: ESR1 and p53 interactome alteration defines mechanisms of tamoxifen response in luminal breast cancer
    Article Snippet: .. After hydration, TMAs were washed twice with tris-buffered saline with Tween 20 (TBST) followed by antigen retrieval using antigen retrieval buffer (EnVision FLEX Target Retrieval Solution, High pH (50×) (Dako Omnis) GV804) in steamer at 100°C for 1 h. TMA slides were cooled at room temperature and further quenched by Duolink hydrogen peroxide (Sigma-DUO82054) for 10 min followed by blocking by Duolink blocking solution (Sigma-DUO82007) for 5 min. Primary antibodies of ERα (HC20-1:100) and p53 (DO1-1:100) were diluted appropriately in antibody dilution buffer applied on each TMA (50 μL) and incubated further for 1 h in a humidified chamber. ..

    Proximity Ligation Assay:

    Article Title: Nonreceptor tyrosine kinase ABL1 regulates lysosomal acidification by phosphorylating the ATP6V1B2 subunit of the vacuolar-type H + -ATPase.
    Article Snippet: The Duolink in situ proximity ligation assay (Sigma-Aldrich, DUO92008) enables the endogenous detection of protein interactions and posttranslational modi cations of proteins at the single-molecule level in xed cell and tissue samples. .. Brie y, A549 cells plated on glass coverslips were xed with 4% formaldehyde, and the cells were permeabilized with 0.2% Triton X-100 in PBS for 15 min. After blocking with Duolink Blocking Solution(Sigma-Aldrich, DUO82007), the cells were incubated with rabbit antiATP6V1B2 (Sigma-Aldrich, HPA00814, 1:100 dilution) and mouse anti-ABL1 (8E9, Santa Cruz Biotechnology, sc -56,887, 1:50 dilution) or mouse anti-phosphotyrosine (4G10, Millipore, 05–321, 1:100 dilution) or mouse anti-ATP6V0D1 (Abcam, ab56441, 1:100 dilution) primary antibodies and then incubated with oligonucleotide-labeled anti-mouse IgG (Sigma-Aldrich, DUO92004) and anti-rabbit IgG (Sigma-Aldrich, DUO92002) according to the manufacturer’s instructions for PLA. ..

    other:

    Article Title: Inhibition of mRNA nuclear export promotes SARS-CoV-2 pathogenesis
    Article Snippet: Samples were then rinsed briefly with PBS and incubated with the Duolink blocking solution (Duolink In Situ Red Starter Mouse/Rabbit kit, DUO9210, Sigma-Aldrich Co. LLC) in a preheated humidity chamber for 30 min at 37 °C.

    Saline:

    Article Title: ESR1 and p53 interactome alteration defines mechanisms of tamoxifen response in luminal breast cancer
    Article Snippet: .. After hydration, TMAs were washed twice with tris-buffered saline with Tween 20 (TBST) followed by antigen retrieval using antigen retrieval buffer (EnVision FLEX Target Retrieval Solution, High pH (50×) (Dako Omnis) GV804) in steamer at 100°C for 1 h. TMA slides were cooled at room temperature and further quenched by Duolink hydrogen peroxide (Sigma-DUO82054) for 10 min followed by blocking by Duolink blocking solution (Sigma-DUO82007) for 5 min. Primary antibodies of ERα (HC20-1:100) and p53 (DO1-1:100) were diluted appropriately in antibody dilution buffer applied on each TMA (50 μL) and incubated further for 1 h in a humidified chamber. ..



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    Physical interaction of CMTM6 <t>with</t> <t>EGFR</t> in NSCLC cells. A) Identification of EGFR‐interacting proteins by mass spectrometry. Cellular extracts from HEK293T cells stably expressing FLAG‐tagged EGFR/L858R were subjected to affinity purification with anti‐FLAG affinity columns. The bound proteins were eluted with FLAG peptides, resolved by SDS‐PAGE, and visualized by silver staining. Protein bands of interest were retrieved and analyzed by mass spectrometry. B) Immunoblotting analysis of the purified eluates. Purified eluates from Figure were analyzed by immunoblotting using antibodies specific to the indicated proteins. C) Co‐immunoprecipitation in PC‐9, HCC827, NCI‐H1975, A549, and NCI‐H1299 cells with anti‐EGFR followed by immunoblotting with antibodies against the indicated proteins. D) <t>Duolink</t> proximity ligation assay (PLA) in PC‐9, HCC827, NCI‐H1975, A549, and NCI‐H1299 cells was performed using anti‐EGFR and anti‐CMTM6 antibodies. The PLA signals (green dots) indicate close proximity between the two proteins. Scale bar, 20 µm. E) GST pull‐down assays were performed with GST or with GST‐fused CMTM6 and in vitro transcribed/translated EGFR (wild‐type and variants bearing NSCLC patient mutations). Coomassie brilliant blue staining of the GST and GST‐fused CMTM6 are shown with arrows (left). The reaction was analyzed by immunoblotting with antibodies against EGFR (right).
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    Physical interaction of CMTM6 with EGFR in NSCLC cells. A) Identification of EGFR‐interacting proteins by mass spectrometry. Cellular extracts from HEK293T cells stably expressing FLAG‐tagged EGFR/L858R were subjected to affinity purification with anti‐FLAG affinity columns. The bound proteins were eluted with FLAG peptides, resolved by SDS‐PAGE, and visualized by silver staining. Protein bands of interest were retrieved and analyzed by mass spectrometry. B) Immunoblotting analysis of the purified eluates. Purified eluates from Figure were analyzed by immunoblotting using antibodies specific to the indicated proteins. C) Co‐immunoprecipitation in PC‐9, HCC827, NCI‐H1975, A549, and NCI‐H1299 cells with anti‐EGFR followed by immunoblotting with antibodies against the indicated proteins. D) Duolink proximity ligation assay (PLA) in PC‐9, HCC827, NCI‐H1975, A549, and NCI‐H1299 cells was performed using anti‐EGFR and anti‐CMTM6 antibodies. The PLA signals (green dots) indicate close proximity between the two proteins. Scale bar, 20 µm. E) GST pull‐down assays were performed with GST or with GST‐fused CMTM6 and in vitro transcribed/translated EGFR (wild‐type and variants bearing NSCLC patient mutations). Coomassie brilliant blue staining of the GST and GST‐fused CMTM6 are shown with arrows (left). The reaction was analyzed by immunoblotting with antibodies against EGFR (right).

    Journal: Advanced Science

    Article Title: A CMTM6 Nanobody Overcomes EGFR‐TKI Resistance in Non‐Small Cell Lung Cancer

    doi: 10.1002/advs.202410945

    Figure Lengend Snippet: Physical interaction of CMTM6 with EGFR in NSCLC cells. A) Identification of EGFR‐interacting proteins by mass spectrometry. Cellular extracts from HEK293T cells stably expressing FLAG‐tagged EGFR/L858R were subjected to affinity purification with anti‐FLAG affinity columns. The bound proteins were eluted with FLAG peptides, resolved by SDS‐PAGE, and visualized by silver staining. Protein bands of interest were retrieved and analyzed by mass spectrometry. B) Immunoblotting analysis of the purified eluates. Purified eluates from Figure were analyzed by immunoblotting using antibodies specific to the indicated proteins. C) Co‐immunoprecipitation in PC‐9, HCC827, NCI‐H1975, A549, and NCI‐H1299 cells with anti‐EGFR followed by immunoblotting with antibodies against the indicated proteins. D) Duolink proximity ligation assay (PLA) in PC‐9, HCC827, NCI‐H1975, A549, and NCI‐H1299 cells was performed using anti‐EGFR and anti‐CMTM6 antibodies. The PLA signals (green dots) indicate close proximity between the two proteins. Scale bar, 20 µm. E) GST pull‐down assays were performed with GST or with GST‐fused CMTM6 and in vitro transcribed/translated EGFR (wild‐type and variants bearing NSCLC patient mutations). Coomassie brilliant blue staining of the GST and GST‐fused CMTM6 are shown with arrows (left). The reaction was analyzed by immunoblotting with antibodies against EGFR (right).

    Article Snippet: PC‐9 cell slides were fixed in 100% methanol at −20 °C for 10 min. After incubation with Duolink blocking solution at 37 °C for 1 h, slides were treated with anti‐EGFR (Abcam) and anti‐CMTM6 (Cell Signaling Technology) primary antibodies overnight at 4 °C.

    Techniques: Mass Spectrometry, Stable Transfection, Expressing, Affinity Purification, SDS Page, Silver Staining, Western Blot, Purification, Immunoprecipitation, Proximity Ligation Assay, In Vitro, Staining

    An anti‐CMTM6 nanobody decreases EGFR levels by blocking CMTM6 from binding to EGFR in NSCLC Cells. A) Immunoblotting analysis of lysates from PC‐9‐GR and NCI‐H1975‐OR cells treated with control or CMTM6 siRNAs under treatment of DMSO, gefitinib, or osimertinib for 24 h. Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. B) PC‐9‐GR cells were treated with IgG control (Ctrl) or anti‐CMTM6 HA‐tagged nanobodies (1A5, 1E2, 3F11, and 4C2) for 12 h at the concentration of 10 µg mL −1 . Cells were treated with CQ for 4 h before being collected for immunoblotting. Co‐immunoprecipitation with anti‐CMTM6 was followed by immunoblotting with antibodies against the indicated proteins. Nb represents the nanobody. C) Immunoblotting of lysates from PC‐9‐GR and NCI‐H1975‐OR cells treated with control or HA‐tagged nanobodies (1A5, 1E2, 3F11, and 4C2). Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. D) Immunofluorescence to monitor the localization of HA‐tagged 1A5 nanobody. PC‐9 cells were treated with control or nanobody 1A5 for 12 h and were fixed and stained with antibodies against HA. Scale bar, 10 µm. E) Co‐immunoprecipitation in PC‐9‐GR cells treated with control or varying concentrations of HA‐tagged 1A5 nanobody (1, 5, or 10 µg mL −1 ) for 24 h. Immunoprecipitation was performed with anti‐CMTM6 followed by immunoblotting. F) Duolink proximity ligation assay in PC‐9‐GR cells treated with control or nanobody 1A5 for 12 h. The assay was performed with anti‐EGFR and anti‐CMTM6 antibodies. Scale bar, 20 µm. G) Co‐immunoprecipitation in PC‐9‐GR cells treated with control or 10 µg mL −1 HA‐tagged 1A5 nanobody for 6, 12, or 24 h with anti‐CMTM6 followed by immunoblotting. H) Immunoblotting of lysates from PC‐9‐GR cells treated with control or CMTM6 siRNAs were treated with 1 or 10 µg mL −1 1A5 for 6, 12, or 24 h. Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. I) Immunoblotting analysis of lysates from PC‐9‐GR and NCI‐H1975‐OR cells treated with control or CMTM6 siRNAs under treatment of control or 10 µg mL −1 1A5 for 24 h. Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. Data are representative of at least three independent experiments.

    Journal: Advanced Science

    Article Title: A CMTM6 Nanobody Overcomes EGFR‐TKI Resistance in Non‐Small Cell Lung Cancer

    doi: 10.1002/advs.202410945

    Figure Lengend Snippet: An anti‐CMTM6 nanobody decreases EGFR levels by blocking CMTM6 from binding to EGFR in NSCLC Cells. A) Immunoblotting analysis of lysates from PC‐9‐GR and NCI‐H1975‐OR cells treated with control or CMTM6 siRNAs under treatment of DMSO, gefitinib, or osimertinib for 24 h. Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. B) PC‐9‐GR cells were treated with IgG control (Ctrl) or anti‐CMTM6 HA‐tagged nanobodies (1A5, 1E2, 3F11, and 4C2) for 12 h at the concentration of 10 µg mL −1 . Cells were treated with CQ for 4 h before being collected for immunoblotting. Co‐immunoprecipitation with anti‐CMTM6 was followed by immunoblotting with antibodies against the indicated proteins. Nb represents the nanobody. C) Immunoblotting of lysates from PC‐9‐GR and NCI‐H1975‐OR cells treated with control or HA‐tagged nanobodies (1A5, 1E2, 3F11, and 4C2). Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. D) Immunofluorescence to monitor the localization of HA‐tagged 1A5 nanobody. PC‐9 cells were treated with control or nanobody 1A5 for 12 h and were fixed and stained with antibodies against HA. Scale bar, 10 µm. E) Co‐immunoprecipitation in PC‐9‐GR cells treated with control or varying concentrations of HA‐tagged 1A5 nanobody (1, 5, or 10 µg mL −1 ) for 24 h. Immunoprecipitation was performed with anti‐CMTM6 followed by immunoblotting. F) Duolink proximity ligation assay in PC‐9‐GR cells treated with control or nanobody 1A5 for 12 h. The assay was performed with anti‐EGFR and anti‐CMTM6 antibodies. Scale bar, 20 µm. G) Co‐immunoprecipitation in PC‐9‐GR cells treated with control or 10 µg mL −1 HA‐tagged 1A5 nanobody for 6, 12, or 24 h with anti‐CMTM6 followed by immunoblotting. H) Immunoblotting of lysates from PC‐9‐GR cells treated with control or CMTM6 siRNAs were treated with 1 or 10 µg mL −1 1A5 for 6, 12, or 24 h. Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. I) Immunoblotting analysis of lysates from PC‐9‐GR and NCI‐H1975‐OR cells treated with control or CMTM6 siRNAs under treatment of control or 10 µg mL −1 1A5 for 24 h. Cellular extracts were prepared, and immunoblotting was performed with the antibodies against the indicated proteins. Data are representative of at least three independent experiments.

    Article Snippet: PC‐9 cell slides were fixed in 100% methanol at −20 °C for 10 min. After incubation with Duolink blocking solution at 37 °C for 1 h, slides were treated with anti‐EGFR (Abcam) and anti‐CMTM6 (Cell Signaling Technology) primary antibodies overnight at 4 °C.

    Techniques: Blocking Assay, Binding Assay, Western Blot, Control, Concentration Assay, Immunoprecipitation, Immunofluorescence, Staining, Proximity Ligation Assay