rabbit monoclonal anti myc  (Cell Signaling Technology Inc)

 
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    Name:
    Anti biotin D5A7 Rabbit mAb HRP Conjugate
    Description:
    Anti biotin D5A7 Rabbit mAb is conjugated to the carbohydrate groups of horseradish peroxidase HRP via its amine groups This product has been optimized to detect biotinylated primary antibodies HRP conjugated antibodies do not require incubation with a secondary antibody Do not mix this antibody in solution with any Anti rabbit antibody Anti rabbit antibodies will cross react with this antibody and could result in decreased activity of both Anti rabbit and Anti biotin D5A7 Rabbit mAb
    Catalog Number:
    5571
    Price:
    None
    Category:
    Secondary Antibodies
    Source:
    Monoclonal antibody is produced by immunizing animals with a biotinylated protein.
    Reactivity:
    All Species Expected
    Applications:
    Western Blot, other
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc rabbit monoclonal anti myc
    Actinonin is unlikely to inhibit the peptide deformylase of P. falciparum . ( A ) Western blot of the parental line (NF54attB-pCRISPR) and the <t>PDF-myc</t> parasites grown with our without aTC for 24 hr. Induction of the second copy of PfPDF (PDF-myc) with 4 uM aTC results in two bands, the top lighter band representing unprocessed PDF-myc, and the bottom darker band representing processed PDF-myc. PfAldolase is used as a loading control. Induction with 0.125–4 uM aTC results in similar amount of PDF-myc induction (data not shown). ( B ) Western blot for PDF-myc of parasites with or without their apicoplast. An accumulation of unprocessed PDF-myc is observed when the apicoplast is missing, due to loss of the transit peptide cleavage that usually occurs upon import to the apicoplast. This has been shown previously for apicoplast-resident proteins and is consistent apicoplast localization ( Yeh and DeRisi, 2011 ). ( C ) Dose dependent parasites growth inhibition by actinonin in the presence of 4 uM aTC does not change the actinonin EC 50 . This experiment was also performed under IPP rescue conditions, to confirm apicoplast specificity of actinonin and with a range of aTC concentrations (0.125–4 uM) to insure max expression of PDF-myc (data not shown). Error bars represent the SEM of 3 biological replicates. ( D ) Parasite growth after one or two replication cycles after treatment with actinonin, chloramphenicol, or both actinonin and chloramphenicol normalized to growth of an untreated control. Treatment with actinonin alone inhibited growth after the first replication cycle, whereas treatment with chloramphenicol alone inhibited growth after the second replication cycle. Co-treatment with chloramphenicol, which targets apicoplast translation, did not suppress effects of actinonin treatment, which was inconsistent with actinonin targeting the peptide deformylase (PDF) of the apicoplast. This experiment was tried using a range of concentrations of actinonin and chloramphenicol to insure the data was not the result of partial inhibition. All concentrations that lead to apicoplast-specific death gave this phenotype (data not shown).
    Anti biotin D5A7 Rabbit mAb is conjugated to the carbohydrate groups of horseradish peroxidase HRP via its amine groups This product has been optimized to detect biotinylated primary antibodies HRP conjugated antibodies do not require incubation with a secondary antibody Do not mix this antibody in solution with any Anti rabbit antibody Anti rabbit antibodies will cross react with this antibody and could result in decreased activity of both Anti rabbit and Anti biotin D5A7 Rabbit mAb
    https://www.bioz.com/result/rabbit monoclonal anti myc/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti myc - by Bioz Stars, 2021-09
    97/100 stars

    Images

    1) Product Images from "Small molecule inhibition of apicomplexan FtsH1 disrupts plastid biogenesis in human pathogens"

    Article Title: Small molecule inhibition of apicomplexan FtsH1 disrupts plastid biogenesis in human pathogens

    Journal: eLife

    doi: 10.7554/eLife.29865

    Actinonin is unlikely to inhibit the peptide deformylase of P. falciparum . ( A ) Western blot of the parental line (NF54attB-pCRISPR) and the PDF-myc parasites grown with our without aTC for 24 hr. Induction of the second copy of PfPDF (PDF-myc) with 4 uM aTC results in two bands, the top lighter band representing unprocessed PDF-myc, and the bottom darker band representing processed PDF-myc. PfAldolase is used as a loading control. Induction with 0.125–4 uM aTC results in similar amount of PDF-myc induction (data not shown). ( B ) Western blot for PDF-myc of parasites with or without their apicoplast. An accumulation of unprocessed PDF-myc is observed when the apicoplast is missing, due to loss of the transit peptide cleavage that usually occurs upon import to the apicoplast. This has been shown previously for apicoplast-resident proteins and is consistent apicoplast localization ( Yeh and DeRisi, 2011 ). ( C ) Dose dependent parasites growth inhibition by actinonin in the presence of 4 uM aTC does not change the actinonin EC 50 . This experiment was also performed under IPP rescue conditions, to confirm apicoplast specificity of actinonin and with a range of aTC concentrations (0.125–4 uM) to insure max expression of PDF-myc (data not shown). Error bars represent the SEM of 3 biological replicates. ( D ) Parasite growth after one or two replication cycles after treatment with actinonin, chloramphenicol, or both actinonin and chloramphenicol normalized to growth of an untreated control. Treatment with actinonin alone inhibited growth after the first replication cycle, whereas treatment with chloramphenicol alone inhibited growth after the second replication cycle. Co-treatment with chloramphenicol, which targets apicoplast translation, did not suppress effects of actinonin treatment, which was inconsistent with actinonin targeting the peptide deformylase (PDF) of the apicoplast. This experiment was tried using a range of concentrations of actinonin and chloramphenicol to insure the data was not the result of partial inhibition. All concentrations that lead to apicoplast-specific death gave this phenotype (data not shown).
    Figure Legend Snippet: Actinonin is unlikely to inhibit the peptide deformylase of P. falciparum . ( A ) Western blot of the parental line (NF54attB-pCRISPR) and the PDF-myc parasites grown with our without aTC for 24 hr. Induction of the second copy of PfPDF (PDF-myc) with 4 uM aTC results in two bands, the top lighter band representing unprocessed PDF-myc, and the bottom darker band representing processed PDF-myc. PfAldolase is used as a loading control. Induction with 0.125–4 uM aTC results in similar amount of PDF-myc induction (data not shown). ( B ) Western blot for PDF-myc of parasites with or without their apicoplast. An accumulation of unprocessed PDF-myc is observed when the apicoplast is missing, due to loss of the transit peptide cleavage that usually occurs upon import to the apicoplast. This has been shown previously for apicoplast-resident proteins and is consistent apicoplast localization ( Yeh and DeRisi, 2011 ). ( C ) Dose dependent parasites growth inhibition by actinonin in the presence of 4 uM aTC does not change the actinonin EC 50 . This experiment was also performed under IPP rescue conditions, to confirm apicoplast specificity of actinonin and with a range of aTC concentrations (0.125–4 uM) to insure max expression of PDF-myc (data not shown). Error bars represent the SEM of 3 biological replicates. ( D ) Parasite growth after one or two replication cycles after treatment with actinonin, chloramphenicol, or both actinonin and chloramphenicol normalized to growth of an untreated control. Treatment with actinonin alone inhibited growth after the first replication cycle, whereas treatment with chloramphenicol alone inhibited growth after the second replication cycle. Co-treatment with chloramphenicol, which targets apicoplast translation, did not suppress effects of actinonin treatment, which was inconsistent with actinonin targeting the peptide deformylase (PDF) of the apicoplast. This experiment was tried using a range of concentrations of actinonin and chloramphenicol to insure the data was not the result of partial inhibition. All concentrations that lead to apicoplast-specific death gave this phenotype (data not shown).

    Techniques Used: Western Blot, Inhibition, Expressing

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    Incubation:

    Article Title: Novel Isoquinoline Alkaloid Litcubanine A - A Potential Anti-Inflammatory Candidate
    Article Snippet: .. Biotinylated secondary antibodies were added and incubated at room temperature for 1 h, and finally, the horseradish peroxidase complex was added with the diaminobenzidine substrate for visualization. ..

    Article Title: WWP1 deficiency protects from cardiac remodeling induced by simulated microgravity
    Article Snippet: .. After three times washes in PBS, biotinylated secondary antibodies were then added and incubated for 1 h at room temperature, followed by color development with DAB kit (ZSGB-bio). ..

    Article Title: S-Nitrosylation of RhoGAP Myosin9A Is Altered in Advanced Diabetic Kidney Disease
    Article Snippet: .. Proteins were resolved by SDS-PAGE in 10% or 4–20% SDS–polyacrylamide gels (BioRad), transferred to polyvinylidene difluoride membranes, blocked with 5% dry-milk or 5% BSA in TBST and incubated with primary antibodies: actin (A2066, Sigma), Myo9A (Abnova, clone 4C11) and RhoA (67B9,Cell Signaling), followed by appropriate species specific HRP-conjugated secondary antibodies (Jackson Immuno Research Laboratories Inc.). ..

    Article Title: Novel fragile X syndrome 2D and 3D brain models based on human isogenic FMRP-KO iPSCs
    Article Snippet: .. Cells were then incubated overnight at 4 °C with primary antibodies at the following dilutions: mouse anti-PAX6 (sc81649 Santa Cruz Biotechnology, 1:50), mouse anti-GFAP (MAB360 Merck Millipore, 1:500), chicken anti-MAP2 (ab5392 Abcam, 1:2000), rabbit anti-β-TUBULIN III (TUJ1) (T2200 Sigma Aldrich, 1:2000), rabbit anti-FMRP (4317 Cell Signaling, 1:50), mouse anti-VGLUT1 (135303 Synaptic Systems, 1:250), rabbit anti-PSD95 (3450 Cell Signaling, 1:250), mouse anti-GAD67 (sc-28376 Santa Cruz, 1:250). ..

    Article Title: Consumption of barley ameliorates the diabetic steatohepatitis and reduces the high transforming growth factor β expression in mice grown in α-minimum essential medium in vitro as embryos
    Article Snippet: .. The membranes were washed again with PBS-T three times, then incubated with horseradish peroxidase-linked anti-biotin antibody (1:2000; #7075, Cell Signaling Technology) in PBS-T containing skim milk at 4 °C overnight. ..

    SDS Page:

    Article Title: S-Nitrosylation of RhoGAP Myosin9A Is Altered in Advanced Diabetic Kidney Disease
    Article Snippet: .. Proteins were resolved by SDS-PAGE in 10% or 4–20% SDS–polyacrylamide gels (BioRad), transferred to polyvinylidene difluoride membranes, blocked with 5% dry-milk or 5% BSA in TBST and incubated with primary antibodies: actin (A2066, Sigma), Myo9A (Abnova, clone 4C11) and RhoA (67B9,Cell Signaling), followed by appropriate species specific HRP-conjugated secondary antibodies (Jackson Immuno Research Laboratories Inc.). ..

    Produced:

    Article Title: Novel Pan-Pim Kinase Inhibitors With Imidazopyridazine and Thiazolidinedione Structure Exert Potent Antitumor Activities
    Article Snippet: .. AntibodiesThe following antibodies were used for immunoblotting: Anti-phospho-Bad (pSer112) (#SAB4300050, Sigma-Aldrich; Merck KgaA, Darmstadt, Germany), Anti-BAD antibody produced in rabbit (#SAB3500336, Sigma-Aldrich; Merck KgaA), Phospho-4E-BP1 (Ser65) (#9451, Cell Signaling Technology, Danvers, MA, USA), 4E-BP1 antibody (#9452, Cell Signaling Technology), p-p21 (Thr145)-R Antibody (#sc-20220-R, Santa Cruz Biotechnology, Santa Cruz, CA, United States), p21 Antibody (C-19) (#sc-397, Santa Cruz Biotechnology), Monoclonal Anti-α-tubulin produced in mouse (#T9026, Sigma-Aldrich; Merck KgaA), and Monoclonal Anti-β-Actin antibody produced in mouse (#A5316, Sigma-Aldrich; Merck KgaA). ..

    other:

    Article Title: Joint degeneration in a mouse model of pseudoachondroplasia: ER stress, inflammation and autophagy blockage
    Article Snippet: Species specific biotinylated secondary antibodies were used for 1 hr at RT.

    Western Blot:

    Article Title: Phloretin suppresses neuroinflammation by autophagy-mediated Nrf2 activation in macrophages
    Article Snippet: .. The following antibodies were used for western blot: mouse anti-β-actin (1:10,000; sc-47778, Santa Cruz Biotechnology), mouse anti-GAPDH (1:10 000; AB_2537659, Invitrogen), rabbit anti-AMPK (1:1000; 5831S, Cell Signaling Technology), rabbit anti-phosphorylated AMPK (1:1000; 2535S, Cell Signaling Technology), rabbit anti-LC3 (1:1000; L7543, Sigma-Aldrich), rabbit anti-p62 (1:1000; 23214, Cell Signaling Technology). ..

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  • 97
    Cell Signaling Technology Inc rabbit monoclonal anti myc
    Actinonin is unlikely to inhibit the peptide deformylase of P. falciparum . ( A ) Western blot of the parental line (NF54attB-pCRISPR) and the <t>PDF-myc</t> parasites grown with our without aTC for 24 hr. Induction of the second copy of PfPDF (PDF-myc) with 4 uM aTC results in two bands, the top lighter band representing unprocessed PDF-myc, and the bottom darker band representing processed PDF-myc. PfAldolase is used as a loading control. Induction with 0.125–4 uM aTC results in similar amount of PDF-myc induction (data not shown). ( B ) Western blot for PDF-myc of parasites with or without their apicoplast. An accumulation of unprocessed PDF-myc is observed when the apicoplast is missing, due to loss of the transit peptide cleavage that usually occurs upon import to the apicoplast. This has been shown previously for apicoplast-resident proteins and is consistent apicoplast localization ( Yeh and DeRisi, 2011 ). ( C ) Dose dependent parasites growth inhibition by actinonin in the presence of 4 uM aTC does not change the actinonin EC 50 . This experiment was also performed under IPP rescue conditions, to confirm apicoplast specificity of actinonin and with a range of aTC concentrations (0.125–4 uM) to insure max expression of PDF-myc (data not shown). Error bars represent the SEM of 3 biological replicates. ( D ) Parasite growth after one or two replication cycles after treatment with actinonin, chloramphenicol, or both actinonin and chloramphenicol normalized to growth of an untreated control. Treatment with actinonin alone inhibited growth after the first replication cycle, whereas treatment with chloramphenicol alone inhibited growth after the second replication cycle. Co-treatment with chloramphenicol, which targets apicoplast translation, did not suppress effects of actinonin treatment, which was inconsistent with actinonin targeting the peptide deformylase (PDF) of the apicoplast. This experiment was tried using a range of concentrations of actinonin and chloramphenicol to insure the data was not the result of partial inhibition. All concentrations that lead to apicoplast-specific death gave this phenotype (data not shown).
    Rabbit Monoclonal Anti Myc, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti myc/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti myc - by Bioz Stars, 2021-09
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    94
    Cell Signaling Technology Inc rabbit anti myc tag monoclonal antibody
    Expression and subcellular localization of SVSP454 and its prey proteins (CCDC181 and MRPL30) in HEK293T cells. SVSP454 (4 µg) and its potential interacting proteins-CCDC181 (4 µg) and MRPL30 (4 µg) were individually a or pairwise (SVSP454 (2 µg)-CCDC181 (2 µg) and SVSP454 (2 µg)-MRPL30 (2 µg)), b transfected into HEK293T cells, stained with a mouse <t>anti-MYC</t> tag monoclonal antibody or rabbit <t>anti-FLAG</t> tag monoclonal antibody, and incubated with a goat anti-mouse Alexa Fluor 488- or donkey anti-rabbit 594-conjugated antibody. Cell nucleus was stained with Hoechst 33342
    Rabbit Anti Myc Tag Monoclonal Antibody, supplied by Cell Signaling Technology Inc, 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/rabbit anti myc tag monoclonal antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti myc tag monoclonal antibody - by Bioz Stars, 2021-09
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    95
    Cell Signaling Technology Inc rabbit anti myc tag
    (a-c) Subcellular localization of SARS-CoV-2 ORF9b. HeLa cells seeded on 12 well coverslips were transfected with the indicated plasmids. After transfection for 20 h, HeLa cells were subject to immunofluorescence staining with mouse <t>anti-Myc</t> antibody and the <t>rabbit</t> antibodies against the corresponding organelle marker. Scale bar, 10 μm. (d) Relative localization of SARS-CoV-2 ORF9b protein with signaling molecules, including RIG-I, MDA5, MAVS, TBK1, TRIF, and STING. The seeding and transfection of HeLa cells were performed as same as in A). After transfection, ORF9b was stained with a rabbit anti-Myc antibody, and the signaling molecules were reacted with mouse antibodies against the indicated tags. Scale bar, 10 μm. TOM20, Mitochondria marker; Calnerxin, ER marker; GM130, Golgi marker. SARS-CoV-2 ORF9b protein, ORF9b.
    Rabbit Anti Myc Tag, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti myc tag/product/Cell Signaling Technology Inc
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    97
    Cell Signaling Technology Inc anti c myc
    BTB reduces ER α , c-Myc, and <t>Cyclin</t> D1 protein expression levels in MCF-7, Ishikawa, and SKOV-3 cells but has no effect on ER α gene expression at mRNA level. Western blot analyses of ER α , c-Myc, and Cyclin D1 levels in control and 2.5 uM BTB treated MCF-7, Ishikawa, and SKOV-3 cells in the absence or presence of 10 nM E2. 50 μ g of total protein from cells was applied onto a 10% sodium dodecyl sulfate-polyacrylamide gel and subjected to electrophoresis followed by Western blot using anti-ER α , <t>anti-c-Myc,</t> or anti-Cyclin D1 antibodies. Representative graphs were shown from consistent results collected from three independent experiments.
    Anti C Myc, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti c myc/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
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    Image Search Results


    Actinonin is unlikely to inhibit the peptide deformylase of P. falciparum . ( A ) Western blot of the parental line (NF54attB-pCRISPR) and the PDF-myc parasites grown with our without aTC for 24 hr. Induction of the second copy of PfPDF (PDF-myc) with 4 uM aTC results in two bands, the top lighter band representing unprocessed PDF-myc, and the bottom darker band representing processed PDF-myc. PfAldolase is used as a loading control. Induction with 0.125–4 uM aTC results in similar amount of PDF-myc induction (data not shown). ( B ) Western blot for PDF-myc of parasites with or without their apicoplast. An accumulation of unprocessed PDF-myc is observed when the apicoplast is missing, due to loss of the transit peptide cleavage that usually occurs upon import to the apicoplast. This has been shown previously for apicoplast-resident proteins and is consistent apicoplast localization ( Yeh and DeRisi, 2011 ). ( C ) Dose dependent parasites growth inhibition by actinonin in the presence of 4 uM aTC does not change the actinonin EC 50 . This experiment was also performed under IPP rescue conditions, to confirm apicoplast specificity of actinonin and with a range of aTC concentrations (0.125–4 uM) to insure max expression of PDF-myc (data not shown). Error bars represent the SEM of 3 biological replicates. ( D ) Parasite growth after one or two replication cycles after treatment with actinonin, chloramphenicol, or both actinonin and chloramphenicol normalized to growth of an untreated control. Treatment with actinonin alone inhibited growth after the first replication cycle, whereas treatment with chloramphenicol alone inhibited growth after the second replication cycle. Co-treatment with chloramphenicol, which targets apicoplast translation, did not suppress effects of actinonin treatment, which was inconsistent with actinonin targeting the peptide deformylase (PDF) of the apicoplast. This experiment was tried using a range of concentrations of actinonin and chloramphenicol to insure the data was not the result of partial inhibition. All concentrations that lead to apicoplast-specific death gave this phenotype (data not shown).

    Journal: eLife

    Article Title: Small molecule inhibition of apicomplexan FtsH1 disrupts plastid biogenesis in human pathogens

    doi: 10.7554/eLife.29865

    Figure Lengend Snippet: Actinonin is unlikely to inhibit the peptide deformylase of P. falciparum . ( A ) Western blot of the parental line (NF54attB-pCRISPR) and the PDF-myc parasites grown with our without aTC for 24 hr. Induction of the second copy of PfPDF (PDF-myc) with 4 uM aTC results in two bands, the top lighter band representing unprocessed PDF-myc, and the bottom darker band representing processed PDF-myc. PfAldolase is used as a loading control. Induction with 0.125–4 uM aTC results in similar amount of PDF-myc induction (data not shown). ( B ) Western blot for PDF-myc of parasites with or without their apicoplast. An accumulation of unprocessed PDF-myc is observed when the apicoplast is missing, due to loss of the transit peptide cleavage that usually occurs upon import to the apicoplast. This has been shown previously for apicoplast-resident proteins and is consistent apicoplast localization ( Yeh and DeRisi, 2011 ). ( C ) Dose dependent parasites growth inhibition by actinonin in the presence of 4 uM aTC does not change the actinonin EC 50 . This experiment was also performed under IPP rescue conditions, to confirm apicoplast specificity of actinonin and with a range of aTC concentrations (0.125–4 uM) to insure max expression of PDF-myc (data not shown). Error bars represent the SEM of 3 biological replicates. ( D ) Parasite growth after one or two replication cycles after treatment with actinonin, chloramphenicol, or both actinonin and chloramphenicol normalized to growth of an untreated control. Treatment with actinonin alone inhibited growth after the first replication cycle, whereas treatment with chloramphenicol alone inhibited growth after the second replication cycle. Co-treatment with chloramphenicol, which targets apicoplast translation, did not suppress effects of actinonin treatment, which was inconsistent with actinonin targeting the peptide deformylase (PDF) of the apicoplast. This experiment was tried using a range of concentrations of actinonin and chloramphenicol to insure the data was not the result of partial inhibition. All concentrations that lead to apicoplast-specific death gave this phenotype (data not shown).

    Article Snippet: For anti-PDF immunoblots, membranes were probed with 1:2000 rabbit monoclonal anti-MYC (Cell Signaling Technology 2278S, Danvers, MA), followed by 1:20,000 rabbit polyclonal anti-PfAldolase (Abcam ab207494, UK) and 1:10,000 donkey anti-rabbit 800 (LiCor Biosciences).

    Techniques: Western Blot, Inhibition, Expressing

    Expression and subcellular localization of SVSP454 and its prey proteins (CCDC181 and MRPL30) in HEK293T cells. SVSP454 (4 µg) and its potential interacting proteins-CCDC181 (4 µg) and MRPL30 (4 µg) were individually a or pairwise (SVSP454 (2 µg)-CCDC181 (2 µg) and SVSP454 (2 µg)-MRPL30 (2 µg)), b transfected into HEK293T cells, stained with a mouse anti-MYC tag monoclonal antibody or rabbit anti-FLAG tag monoclonal antibody, and incubated with a goat anti-mouse Alexa Fluor 488- or donkey anti-rabbit 594-conjugated antibody. Cell nucleus was stained with Hoechst 33342

    Journal: Parasites & Vectors

    Article Title: Screening and identification of Theileria annulata subtelomere-encoded variable secreted protein-950454 (SVSP454) interacting proteins from bovine B cells

    doi: 10.1186/s13071-021-04820-4

    Figure Lengend Snippet: Expression and subcellular localization of SVSP454 and its prey proteins (CCDC181 and MRPL30) in HEK293T cells. SVSP454 (4 µg) and its potential interacting proteins-CCDC181 (4 µg) and MRPL30 (4 µg) were individually a or pairwise (SVSP454 (2 µg)-CCDC181 (2 µg) and SVSP454 (2 µg)-MRPL30 (2 µg)), b transfected into HEK293T cells, stained with a mouse anti-MYC tag monoclonal antibody or rabbit anti-FLAG tag monoclonal antibody, and incubated with a goat anti-mouse Alexa Fluor 488- or donkey anti-rabbit 594-conjugated antibody. Cell nucleus was stained with Hoechst 33342

    Article Snippet: After washing three times with TBST buffer for 5 min each time, primary antibodies that were mouse anti-FLAG tag monoclonal antibody (Sigma, #F1804) or rabbit anti-MYC tag monoclonal antibody (CST, #2278S) were added and incubated overnight at 4 °C.

    Techniques: Expressing, Transfection, Staining, FLAG-tag, Incubation

    SVSP454 interacts with both CCDC181 and MRPL30. Coimmunoprecipitation (Co-IP) and immunoblotting (IB) identification of HEK293T cells expressing SVSP454-MYC (10 µg) and its potential interacting protein FLAG-tagged CCDC181 (10 µg) ( a ) and SVSP454-MYC (10 µg) along with FLAG-tagged MRPL30 (10 µg) ( b ). WCL represents IB analysis of whole HEK293T cell lysates without IP

    Journal: Parasites & Vectors

    Article Title: Screening and identification of Theileria annulata subtelomere-encoded variable secreted protein-950454 (SVSP454) interacting proteins from bovine B cells

    doi: 10.1186/s13071-021-04820-4

    Figure Lengend Snippet: SVSP454 interacts with both CCDC181 and MRPL30. Coimmunoprecipitation (Co-IP) and immunoblotting (IB) identification of HEK293T cells expressing SVSP454-MYC (10 µg) and its potential interacting protein FLAG-tagged CCDC181 (10 µg) ( a ) and SVSP454-MYC (10 µg) along with FLAG-tagged MRPL30 (10 µg) ( b ). WCL represents IB analysis of whole HEK293T cell lysates without IP

    Article Snippet: After washing three times with TBST buffer for 5 min each time, primary antibodies that were mouse anti-FLAG tag monoclonal antibody (Sigma, #F1804) or rabbit anti-MYC tag monoclonal antibody (CST, #2278S) were added and incubated overnight at 4 °C.

    Techniques: Co-Immunoprecipitation Assay, Expressing

    (a-c) Subcellular localization of SARS-CoV-2 ORF9b. HeLa cells seeded on 12 well coverslips were transfected with the indicated plasmids. After transfection for 20 h, HeLa cells were subject to immunofluorescence staining with mouse anti-Myc antibody and the rabbit antibodies against the corresponding organelle marker. Scale bar, 10 μm. (d) Relative localization of SARS-CoV-2 ORF9b protein with signaling molecules, including RIG-I, MDA5, MAVS, TBK1, TRIF, and STING. The seeding and transfection of HeLa cells were performed as same as in A). After transfection, ORF9b was stained with a rabbit anti-Myc antibody, and the signaling molecules were reacted with mouse antibodies against the indicated tags. Scale bar, 10 μm. TOM20, Mitochondria marker; Calnerxin, ER marker; GM130, Golgi marker. SARS-CoV-2 ORF9b protein, ORF9b.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 ORF9b Antagonizes Type I and III Interferons by Targeting Multiple Components of RIG-I/MDA-5-MAVS, TLR3-TRIF, and cGAS-STING Signaling Pathways

    doi: 10.1101/2020.08.16.252973

    Figure Lengend Snippet: (a-c) Subcellular localization of SARS-CoV-2 ORF9b. HeLa cells seeded on 12 well coverslips were transfected with the indicated plasmids. After transfection for 20 h, HeLa cells were subject to immunofluorescence staining with mouse anti-Myc antibody and the rabbit antibodies against the corresponding organelle marker. Scale bar, 10 μm. (d) Relative localization of SARS-CoV-2 ORF9b protein with signaling molecules, including RIG-I, MDA5, MAVS, TBK1, TRIF, and STING. The seeding and transfection of HeLa cells were performed as same as in A). After transfection, ORF9b was stained with a rabbit anti-Myc antibody, and the signaling molecules were reacted with mouse antibodies against the indicated tags. Scale bar, 10 μm. TOM20, Mitochondria marker; Calnerxin, ER marker; GM130, Golgi marker. SARS-CoV-2 ORF9b protein, ORF9b.

    Article Snippet: Rabbit anti-Myc-tag (71D10), rabbit anti-IRF3 (D83B9), rabbit anti-pIRF3 (4D46), rabbit anti-TBK1 (3031S), rabbit anti-pTBK1 (D52C2) were purchased from Cell Signaling Technology; Mouse anti-MAVS was purchased from Santa Cruz; Mouse anti-actin, mouse anti-V5-tag, and rabbit anti-calnexin were purchased from proteintech; Mouse anti-Flag M2 was purchased from Sigma Aldrich; Mouse anti-Myc-tag (9E10) was purchased from Origene; Rabbit anti-GM130 was purchased from Abcam; Rabbit anti-Tom20 antibody was purchased from Abclonal; Mouse anti-HA was purchased from MDL biotech.

    Techniques: Transfection, Immunofluorescence, Staining, Marker

    SARS-CoV-2 ORF9b suppressed IRF3 phosphorylation and nuclear translocation. ( a ) HeLa cells were seeded on 12 well coverslips (5*10^4 cells per well) one day before transfection. HeLa cells were further subjected to infection by SeV after transfection with the Myc vector plasmid or Myc-ORF9b plasmids for 20 h. Following infection for 8h, the slides were harvested and processed for immunofluorescence staining with mouse anti-Myc antibody and rabbit anti-IRF3 antibody. ( b ) Quantification of the percentage of IRF3 in the nucleus upon SeV infection. IRF3 localization from 50 cells within each group was counted and calculated before and after SeV infection. ( c ) SARS-CoV-2 ORF9b protein affects the phosphorylation of IRF3 upon SeV infection. HeLa cells seeded on 6 well plates (5*10^5 cells per well) were transfected with the Myc vector plasmid or Myc-ORF9b plasmid for 20 h before infection with SeV. At the indicated time points, cells were scraped and processed for immunoblotting with the indicated antibodies. SARS-CoV-2 ORF9b protein, ORF9b.

    Journal: bioRxiv

    Article Title: SARS-CoV-2 ORF9b Antagonizes Type I and III Interferons by Targeting Multiple Components of RIG-I/MDA-5-MAVS, TLR3-TRIF, and cGAS-STING Signaling Pathways

    doi: 10.1101/2020.08.16.252973

    Figure Lengend Snippet: SARS-CoV-2 ORF9b suppressed IRF3 phosphorylation and nuclear translocation. ( a ) HeLa cells were seeded on 12 well coverslips (5*10^4 cells per well) one day before transfection. HeLa cells were further subjected to infection by SeV after transfection with the Myc vector plasmid or Myc-ORF9b plasmids for 20 h. Following infection for 8h, the slides were harvested and processed for immunofluorescence staining with mouse anti-Myc antibody and rabbit anti-IRF3 antibody. ( b ) Quantification of the percentage of IRF3 in the nucleus upon SeV infection. IRF3 localization from 50 cells within each group was counted and calculated before and after SeV infection. ( c ) SARS-CoV-2 ORF9b protein affects the phosphorylation of IRF3 upon SeV infection. HeLa cells seeded on 6 well plates (5*10^5 cells per well) were transfected with the Myc vector plasmid or Myc-ORF9b plasmid for 20 h before infection with SeV. At the indicated time points, cells were scraped and processed for immunoblotting with the indicated antibodies. SARS-CoV-2 ORF9b protein, ORF9b.

    Article Snippet: Rabbit anti-Myc-tag (71D10), rabbit anti-IRF3 (D83B9), rabbit anti-pIRF3 (4D46), rabbit anti-TBK1 (3031S), rabbit anti-pTBK1 (D52C2) were purchased from Cell Signaling Technology; Mouse anti-MAVS was purchased from Santa Cruz; Mouse anti-actin, mouse anti-V5-tag, and rabbit anti-calnexin were purchased from proteintech; Mouse anti-Flag M2 was purchased from Sigma Aldrich; Mouse anti-Myc-tag (9E10) was purchased from Origene; Rabbit anti-GM130 was purchased from Abcam; Rabbit anti-Tom20 antibody was purchased from Abclonal; Mouse anti-HA was purchased from MDL biotech.

    Techniques: Translocation Assay, Transfection, Infection, Plasmid Preparation, Immunofluorescence, Staining

    BTB reduces ER α , c-Myc, and Cyclin D1 protein expression levels in MCF-7, Ishikawa, and SKOV-3 cells but has no effect on ER α gene expression at mRNA level. Western blot analyses of ER α , c-Myc, and Cyclin D1 levels in control and 2.5 uM BTB treated MCF-7, Ishikawa, and SKOV-3 cells in the absence or presence of 10 nM E2. 50 μ g of total protein from cells was applied onto a 10% sodium dodecyl sulfate-polyacrylamide gel and subjected to electrophoresis followed by Western blot using anti-ER α , anti-c-Myc, or anti-Cyclin D1 antibodies. Representative graphs were shown from consistent results collected from three independent experiments.

    Journal: BioMed Research International

    Article Title: The Wedelolactone Derivative Inhibits Estrogen Receptor-Mediated Breast, Endometrial, and Ovarian Cancer Cells Growth

    doi: 10.1155/2014/713263

    Figure Lengend Snippet: BTB reduces ER α , c-Myc, and Cyclin D1 protein expression levels in MCF-7, Ishikawa, and SKOV-3 cells but has no effect on ER α gene expression at mRNA level. Western blot analyses of ER α , c-Myc, and Cyclin D1 levels in control and 2.5 uM BTB treated MCF-7, Ishikawa, and SKOV-3 cells in the absence or presence of 10 nM E2. 50 μ g of total protein from cells was applied onto a 10% sodium dodecyl sulfate-polyacrylamide gel and subjected to electrophoresis followed by Western blot using anti-ER α , anti-c-Myc, or anti-Cyclin D1 antibodies. Representative graphs were shown from consistent results collected from three independent experiments.

    Article Snippet: The blots were probed with primary anti-ERα (Novocastra, 6F11), anti-c-Myc (cell signaling, #9402), anti-Cyclin D1 (#2922), anti-Stat3 (#9132), anti-pStat3 (#9145), anti-Akt (#9272), and anti-pAkt(S-473) (#9271) antibodies with dilutions of 1 : 500 to 1 : 1,000 and incubated at room temperature for 2 hrs.

    Techniques: Expressing, Western Blot, Electrophoresis