rabbit gapdh polyclonal  (Proteintech)

 
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    Name:
    Rabbit GAPDH Polyclonal
    Description:
    The GAPDH antibody from Proteintech is a rabbit polyclonal antibody to a recombinant protein of human GAPDH This antibody recognizes human mouse rat pig antigen The GAPDH antibody has been validated for the following applications ELISA FC IF IHC IP WB analysis
    Catalog Number:
    10494-1-AP
    Price:
    [219.03]
    Applications:
    IHC,Western Blot,ELISA,Flow Cytometry,Immunoprecipitation,Immunofluorescence
    Host:
    Rabbit
    Conjugate:
    Unconjugated
    Immunogen:
    Recombinant Protein
    Size:
    150 ul
    Category:
    Antibody
    Antibody Type:
    Primary antibody
    Isotype:
    IgG
    Reactivity:
    Human Mouse Pig Rat Zebrafish Monkey Sea cucumber Shrews
    Buy from Supplier


    Structured Review

    Proteintech rabbit gapdh polyclonal
    Western blot detection of divalent metal transporter 1 (DMT1) in Brugia malayi adult females. Soluble and membrane protein was extracted from B. malayi adult females (soluble protein lanes 1 and 6; membrane protein lanes 2 and 7) and HEK293 cells (soluble protein lanes 3 and 8; membrane protein lanes 4 and 9). In addition, a whole cell protein lysate was extracted from human Caco-2 cells (lanes 5 and 10). Approximately 20 μg of protein from each extract was subjected to SDS-PAGE followed by Western blot analysis with rabbit <t>polyclonal</t> anti-DMT1 antibody (Abcam, Cat: ab123085) (lanes 1 to 4) and rabbit polyclonal <t>anti-GAPDH</t> antibody (Proteintech, Cat: 10494-1-AP) (lanes 5 to 8).
    The GAPDH antibody from Proteintech is a rabbit polyclonal antibody to a recombinant protein of human GAPDH This antibody recognizes human mouse rat pig antigen The GAPDH antibody has been validated for the following applications ELISA FC IF IHC IP WB analysis
    https://www.bioz.com/result/rabbit gapdh polyclonal/product/Proteintech
    Average 99 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    rabbit gapdh polyclonal - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Characterization of Divalent Metal Transporter 1 (DMT1) in Brugia malayi suggests an intestinal-associated pathway for iron absorption"

    Article Title: Characterization of Divalent Metal Transporter 1 (DMT1) in Brugia malayi suggests an intestinal-associated pathway for iron absorption

    Journal: International Journal for Parasitology: Drugs and Drug Resistance

    doi: 10.1016/j.ijpddr.2018.06.003

    Western blot detection of divalent metal transporter 1 (DMT1) in Brugia malayi adult females. Soluble and membrane protein was extracted from B. malayi adult females (soluble protein lanes 1 and 6; membrane protein lanes 2 and 7) and HEK293 cells (soluble protein lanes 3 and 8; membrane protein lanes 4 and 9). In addition, a whole cell protein lysate was extracted from human Caco-2 cells (lanes 5 and 10). Approximately 20 μg of protein from each extract was subjected to SDS-PAGE followed by Western blot analysis with rabbit polyclonal anti-DMT1 antibody (Abcam, Cat: ab123085) (lanes 1 to 4) and rabbit polyclonal anti-GAPDH antibody (Proteintech, Cat: 10494-1-AP) (lanes 5 to 8).
    Figure Legend Snippet: Western blot detection of divalent metal transporter 1 (DMT1) in Brugia malayi adult females. Soluble and membrane protein was extracted from B. malayi adult females (soluble protein lanes 1 and 6; membrane protein lanes 2 and 7) and HEK293 cells (soluble protein lanes 3 and 8; membrane protein lanes 4 and 9). In addition, a whole cell protein lysate was extracted from human Caco-2 cells (lanes 5 and 10). Approximately 20 μg of protein from each extract was subjected to SDS-PAGE followed by Western blot analysis with rabbit polyclonal anti-DMT1 antibody (Abcam, Cat: ab123085) (lanes 1 to 4) and rabbit polyclonal anti-GAPDH antibody (Proteintech, Cat: 10494-1-AP) (lanes 5 to 8).

    Techniques Used: Western Blot, SDS Page

    2) Product Images from "Characterization of Divalent Metal Transporter 1 (DMT1) in Brugia malayi suggests an intestinal-associated pathway for iron absorption"

    Article Title: Characterization of Divalent Metal Transporter 1 (DMT1) in Brugia malayi suggests an intestinal-associated pathway for iron absorption

    Journal: International Journal for Parasitology: Drugs and Drug Resistance

    doi: 10.1016/j.ijpddr.2018.06.003

    Western blot detection of divalent metal transporter 1 (DMT1) in Brugia malayi adult females. Soluble and membrane protein was extracted from B. malayi adult females (soluble protein lanes 1 and 6; membrane protein lanes 2 and 7) and HEK293 cells (soluble protein lanes 3 and 8; membrane protein lanes 4 and 9). In addition, a whole cell protein lysate was extracted from human Caco-2 cells (lanes 5 and 10). Approximately 20 μg of protein from each extract was subjected to SDS-PAGE followed by Western blot analysis with rabbit polyclonal anti-DMT1 antibody (Abcam, Cat: ab123085) (lanes 1 to 4) and rabbit polyclonal anti-GAPDH antibody (Proteintech, Cat: 10494-1-AP) (lanes 5 to 8).
    Figure Legend Snippet: Western blot detection of divalent metal transporter 1 (DMT1) in Brugia malayi adult females. Soluble and membrane protein was extracted from B. malayi adult females (soluble protein lanes 1 and 6; membrane protein lanes 2 and 7) and HEK293 cells (soluble protein lanes 3 and 8; membrane protein lanes 4 and 9). In addition, a whole cell protein lysate was extracted from human Caco-2 cells (lanes 5 and 10). Approximately 20 μg of protein from each extract was subjected to SDS-PAGE followed by Western blot analysis with rabbit polyclonal anti-DMT1 antibody (Abcam, Cat: ab123085) (lanes 1 to 4) and rabbit polyclonal anti-GAPDH antibody (Proteintech, Cat: 10494-1-AP) (lanes 5 to 8).

    Techniques Used: Western Blot, SDS Page

    Related Articles

    Western Blot:

    Article Title: Zfp281 is essential for mouse epiblast maturation through transcriptional and epigenetic control of Nodal signaling
    Article Snippet: .. Western blot analysis was carried out using the following primary antibodies: Zfp281 (sc-166933, Santa Cruz, RRID: AB_10612046 ), p-Smad2 (#3108, Cell Signaling, RRID: AB_490941 ), Smad2 (#3103, Cell Signaling, RRID: AB_490816 ), Lefty (sc-365845, Santa Cruz, RRID: AB_10847353 , detecting both Lefty1 and Lefty2), Gapdh (10494–1-AP, ProteinTech, RRID: AB_2263076 ). .. Blot intensities were quantified using ImageJ software and Student T-test was used to examine statistical significance.

    Article Title: Tollip coordinates Parkin‐dependent trafficking of mitochondrial‐derived vesicles
    Article Snippet: .. Antibodies and reagentsPrimary antibodies used for Western blotting were specific against Tollip (GTX116566, 1:1,000) from GeneTex; Tom1 (ab99356, 1:1,000), Parkin (ab77924, 1:500), COXII (MTCO2, ab110258, 1:1,000), PDH E2/E3bp (ab110333, 1:1,000) and Rab7a (ab137029, 1:500) from Abcam; Actin (612656, 1:2,000) and LAMP1 (555798, 1:500) from BD Biosciences; GAPDH (10494‐1‐AP, 1:2,000) from ProteinTech; MFN2 (7581, 1:500), PINK1 (6946, 1:500) and ATG5 (26305, 1:500) from Cell Signalling; TOM20 (SC‐11415, 1:2,000) and VPS35 (SC‐374372, 1:1,000) from Santa Cruz; GFP (A11122, 1:1,000) from Invitrogen; Myc (9E10, MAB3696‐SP, 1:1,000) and K48‐linked ubiquitin (A‐101, 1:20,000) from R & D Systems; and ubiquitin (FK2, BML‐PW8810‐0100, 1:2,000) from Enzo. .. Primary antibodies used from immunofluorescence were specific against Tollip (GTX116566, 1:100) from GeneTex or (ATO0918, 1:100) from Insight Biotechnology Ltd; Tom1 (ab99356, 1:100), Parkin (ab77924, 1:250), PDH E2/E3bp (ab110333, 1:1,000), Rab7a (ab137029, 1:100) and GFP (A11122, 1:1,000) from Abcam; LAMP1 (555798, 1:250), Rab5 (610282, 1:200), EEA1 (610456, 1:250) and GM130 (610823, 1:1,000) from BD Biosciences; HA (901501, 1:500) and Cytochrome c (612302, 1:1,000) from BioLegend; MFN2 (7581, 1:500) and HA (37245, 1:1,000) from Cell Signalling; TOM20 (SC‐11415, 1:1,000) and VPS35 (SC‐374372, 1:1,000) from Santa Cruz; GFP (A11122, 1:2,000) from Invitrogen; Myc (9E10, MAB3696‐SP, 1:1,000) from R & D Systems; ubiquitin (FK2, BML‐PW8810‐0100, 1:1,000) from Enzo; and Cathepsin D (IM16, 1:100) from Oncogene.

    Incubation:

    Article Title: MicroRNA deep sequencing in two adult stem cell populations identifies miR-501 as a novel regulator of myosin heavy chain during muscle regeneration
    Article Snippet: .. Membranes were incubated at 4°C with the primary antibodies for at least 16 h [anti-total myosin heavy chain from DSHB, MF-20 (1:100), anti-myosin heavy chain-3 from Santa Cruz (sc-53091; 1:500), anti-desmin from Sigma (D1033; 1:200), anti-GAPDH from ProteinTech (10494-1-AP; 1:1000), anti-gigaxonin from Proteintech (14305-1-AP; 1:500)]. .. Incubation with appropriate HRP-conjugated secondary antibodies and the Lumi-Light western blotting substrate (Roche) was used to detect the signal in a LAS-3000 imager (Fujifilm).

    Article Title: Down-regulation of miR-30b-5p protects cardiomyocytes against hypoxia-induced injury by targeting Aven
    Article Snippet: .. The membranes were blocked with 5% skim milk diluted in TBS-Tween for 1 h and incubated at 4 °C overnight with anti-Bax (1: 500, #2774, Cell signaling), anti-Bcl-2 (1: 500, #2876, Proteintech), anti-Aven (1: 1000, #2865, Cell signaling) or anti-GAPDH (1: 500000, 10,494–1-AP, Proteintech). .. Next day, the membranes were incubated with HRP-conjugated secondary antibodies (1: 5000, SC-2005, Santa Cruz).

    Article Title: PAK4, a target of miR-9-5p, promotes cell proliferation and inhibits apoptosis in colorectal cancer
    Article Snippet: .. It was then incubated with primary antibodies against PAK4 (1:1000, ab227197; Abcam) and GAPDH (1:5000, 10,494–1-AP; Proteintech) overnight at 4 °C, followed by incubation with horseradish peroxidase-conjugated secondary antibody (1:5000, SC-2054; Santa Cruz Biotechnology). .. The bands of target protein were visualized using an enhanced chemiluminescence reagent (Bio-Rad Laboratories) and quantified using Image-pro plus 6.0 Software.

    Article Title: MiR-30b-5p regulates the lipid metabolism by targeting PPARGC1A in Huh-7 cell line
    Article Snippet: .. The membranes were blocked with 5% BSA at room temperature for 1 h prior to incubation with primary antibodies, which included anti-PPAR-α (Species: rabbit; Company: proteintech; Catalog: 15540–1-AP) (1:1000), anti-SREBP-1 (Species: rabbit; Company: Absin; Catalog: abs131802) (1:500), anti-GLUT1 (Species: rabbit; Company: proteintech; Catalog: 21829–1-AP) (1:1000), anti-GAPDH (Species: rabbit; Company: proteintech; Catalog: 10494–1-AP) (1:1000). .. The membranes were incubated with the primary antibodies overnight at 4 °C and washed 5 times with PBS containing 0.1% Tween-20 (PBST).

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  • 99
    Proteintech gapdh
    Silencing <t>ITGA2</t> suppresses the aggressive ability of malignant cancer in vitro a and b . RT-PCR ( a ) and Western blot analysis ( b ) of ITGA2 expression in PANC-1, HepG2, SGC-7901, and MDA-MB-231 cells infected with sh-Control or sh-ITGA2s. <t>GAPDH</t> served as an internal reference. Data presented as the mean ± SD of three independent experiments. Each sh-ITGA2 group was compared with sh-Control group. Statistical analyses were performed with one-way ANOVA followed by Tukey’s multiple comparison’s tests. **, P
    Gapdh, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gapdh/product/Proteintech
    Average 99 stars, based on 166 article reviews
    Price from $9.99 to $1999.99
    gapdh - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Proteintech anti bax
    <t>Bcl-2</t> overexpression inhibits the activation of the mitochondrial apoptosis pathway by cisplatin in SKOV3 cells. (A and B) Pc-SKOV3 and Bcl-2-SKOV3 cells were treated with 6 µg/ml cisplatin for 6 h and then stained with MitoPotential dye and 7-AAD to assess the Δψm by flow cytometry. (C and D) Western blot analysis of cytochrome c , Bcl-2, <t>Bax,</t> cleaved caspase-9 and cleaved caspase-3 levels in pc-SKOV3 and Bcl-2-SKOV3 cells after cisplatin treatment. The results are the mean ± SD of three independent experiments. *P
    Anti Bax, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti bax/product/Proteintech
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    anti bax - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    90
    Proteintech rabbit polyclonal antibodies against glyceraldehyde 3 phosphate dehydrogenase gapdh
    Lp MOPV promotes the nuclear translocation of IRF3. (A) HEK 293T cells were cotransfected with Flag-IRF3 and pCMV-Myc (Vec), pCMV-Myc-NP LASV (NP), or pCMV-Myc-Lp MOPV (Lp). At 40 h posttransfection, the cells were collected and then fractionated into nuclear and cytoplasmic fractions. Equal amounts of protein from the cytoplasmic fraction and nuclear fraction were separated by SDS-PAGE and transferred to PVDF membranes. The membranes were then probed with antibodies, and the bands were visualized. Band intensities were measured with Quantity One software. To normalize IRF3 protein intensity, <t>GAPDH</t> and lamin B1 were used as the cytoplasmic and nuclear endogenous loading controls, respectively. (B) HeLa cells were cotransfected with Flag-IRF3 and pCMV-Myc (Vec) or pCMV-Myc-Lp MOPV (Lp). At 36 h posttransfection, the cells were fixed and incubated with the anti-Flag rabbit <t>polyclonal</t> antibody, followed by a DyLight488-labeled antibody to rabbit IgG to detect Flag-IRF3 (green) and the anti-Myc mouse monoclonal antibody followed by a Cy5-labeled antibody to mouse IgG to detect Myc-Lp (red). Next, the cells were stained with DAPI (blue) to visualize nuclei. Cells with Flag-IRF3 signals are indicated by arrows. Immunofluorescence analysis was performed with an A1 MP+ multiphoton confocal microscope (Nikon).
    Rabbit Polyclonal Antibodies Against Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Proteintech, 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/result/rabbit polyclonal antibodies against glyceraldehyde 3 phosphate dehydrogenase gapdh/product/Proteintech
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal antibodies against glyceraldehyde 3 phosphate dehydrogenase gapdh - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    Image Search Results


    Silencing ITGA2 suppresses the aggressive ability of malignant cancer in vitro a and b . RT-PCR ( a ) and Western blot analysis ( b ) of ITGA2 expression in PANC-1, HepG2, SGC-7901, and MDA-MB-231 cells infected with sh-Control or sh-ITGA2s. GAPDH served as an internal reference. Data presented as the mean ± SD of three independent experiments. Each sh-ITGA2 group was compared with sh-Control group. Statistical analyses were performed with one-way ANOVA followed by Tukey’s multiple comparison’s tests. **, P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Overexpressed ITGA2 promotes malignant tumor aggression by up-regulating PD-L1 expression through the activation of the STAT3 signaling pathway

    doi: 10.1186/s13046-019-1496-1

    Figure Lengend Snippet: Silencing ITGA2 suppresses the aggressive ability of malignant cancer in vitro a and b . RT-PCR ( a ) and Western blot analysis ( b ) of ITGA2 expression in PANC-1, HepG2, SGC-7901, and MDA-MB-231 cells infected with sh-Control or sh-ITGA2s. GAPDH served as an internal reference. Data presented as the mean ± SD of three independent experiments. Each sh-ITGA2 group was compared with sh-Control group. Statistical analyses were performed with one-way ANOVA followed by Tukey’s multiple comparison’s tests. **, P

    Article Snippet: The ITGA2 antibody (ab133557, 1:1000) was purchased from Abcam; GAPDH (10494–1-AP, 1:3000) from Proteintech; STAT3 (10253–2-AP, 1:1000) from Proteintech; PD-L1 (13684S, 1:1000) from Cell Signaling Technology; and Phospho-STAT3-Y705 (abs118973, 1:1000) from Absin.

    Techniques: In Vitro, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Multiple Displacement Amplification, Infection

    ITGA2 promoted tumor growth in pancreatic cancer in vivo. a and b . PANC-1 cells were infected with sh-Control, sh-ITGA2, or sh-ITGA2 and ITGA2 plasmid. The cells were harvested for RT-PCR analysis ( a ) and Western blot analysis ( b ). GAPDH served as an internal reference. sh-Control group was compared with sh-ITGA2 group, sh-ITGA2 group was compared with sh-ITGA2 + ITGA2 group. Statistical analyses were performed with one-way ANOVA followed by Tukey’s multiple comparison’s tests. ***, P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Overexpressed ITGA2 promotes malignant tumor aggression by up-regulating PD-L1 expression through the activation of the STAT3 signaling pathway

    doi: 10.1186/s13046-019-1496-1

    Figure Lengend Snippet: ITGA2 promoted tumor growth in pancreatic cancer in vivo. a and b . PANC-1 cells were infected with sh-Control, sh-ITGA2, or sh-ITGA2 and ITGA2 plasmid. The cells were harvested for RT-PCR analysis ( a ) and Western blot analysis ( b ). GAPDH served as an internal reference. sh-Control group was compared with sh-ITGA2 group, sh-ITGA2 group was compared with sh-ITGA2 + ITGA2 group. Statistical analyses were performed with one-way ANOVA followed by Tukey’s multiple comparison’s tests. ***, P

    Article Snippet: The ITGA2 antibody (ab133557, 1:1000) was purchased from Abcam; GAPDH (10494–1-AP, 1:3000) from Proteintech; STAT3 (10253–2-AP, 1:1000) from Proteintech; PD-L1 (13684S, 1:1000) from Cell Signaling Technology; and Phospho-STAT3-Y705 (abs118973, 1:1000) from Absin.

    Techniques: In Vivo, Infection, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Overpressed ITGA2 promotes the aggressive ability of malignant cancer in vitro. a and b . RT-PCR ( a ) and Western blot analysis ( b ) of ITGA2 expression in PANC-1, HepG2, SGC-7901, and MDA-MB-231 cells with normal or stably overexpressed ITGA2. GAPDH served as an internal reference. Data presented as the mean ± SD of three independent experiments. ITGA2 overexppression groups were compared with pcDNA 3.1 transfection group. Statistical analyses were performed with one-way ANOVA followed by Tukey’s multiple comparison’s tests. Compared groups were shown in the figures. ***, P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Overexpressed ITGA2 promotes malignant tumor aggression by up-regulating PD-L1 expression through the activation of the STAT3 signaling pathway

    doi: 10.1186/s13046-019-1496-1

    Figure Lengend Snippet: Overpressed ITGA2 promotes the aggressive ability of malignant cancer in vitro. a and b . RT-PCR ( a ) and Western blot analysis ( b ) of ITGA2 expression in PANC-1, HepG2, SGC-7901, and MDA-MB-231 cells with normal or stably overexpressed ITGA2. GAPDH served as an internal reference. Data presented as the mean ± SD of three independent experiments. ITGA2 overexppression groups were compared with pcDNA 3.1 transfection group. Statistical analyses were performed with one-way ANOVA followed by Tukey’s multiple comparison’s tests. Compared groups were shown in the figures. ***, P

    Article Snippet: The ITGA2 antibody (ab133557, 1:1000) was purchased from Abcam; GAPDH (10494–1-AP, 1:3000) from Proteintech; STAT3 (10253–2-AP, 1:1000) from Proteintech; PD-L1 (13684S, 1:1000) from Cell Signaling Technology; and Phospho-STAT3-Y705 (abs118973, 1:1000) from Absin.

    Techniques: In Vitro, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Multiple Displacement Amplification, Stable Transfection, Transfection

    Inhibition of miR-30b-5p suppresses hypoxia-induced apoptosis in cardiomyocytes. a Representative captures of flow-cytometric data demonstrating the percent early apoptosis (Annexin V+/PI-) and late apoptosis (Annexin V+/PI+) in AC16 cells grown under normoxia or hypoxia with or without miR-30b-5p inhibitor. b Quantitation of A. c Protein expression of Bax and Bcl-2 was detected by western blot analysis in AC16 cells grown under normoxia or hypoxia with or without miR-30b-5p inhibitor. GAPDH was used as an internal control. *: hypoxia vs. normoxia; #: hypoxia + inhibitor vs. hypoxia + NC; *** p

    Journal: Cellular & Molecular Biology Letters

    Article Title: Down-regulation of miR-30b-5p protects cardiomyocytes against hypoxia-induced injury by targeting Aven

    doi: 10.1186/s11658-019-0187-4

    Figure Lengend Snippet: Inhibition of miR-30b-5p suppresses hypoxia-induced apoptosis in cardiomyocytes. a Representative captures of flow-cytometric data demonstrating the percent early apoptosis (Annexin V+/PI-) and late apoptosis (Annexin V+/PI+) in AC16 cells grown under normoxia or hypoxia with or without miR-30b-5p inhibitor. b Quantitation of A. c Protein expression of Bax and Bcl-2 was detected by western blot analysis in AC16 cells grown under normoxia or hypoxia with or without miR-30b-5p inhibitor. GAPDH was used as an internal control. *: hypoxia vs. normoxia; #: hypoxia + inhibitor vs. hypoxia + NC; *** p

    Article Snippet: The membranes were blocked with 5% skim milk diluted in TBS-Tween for 1 h and incubated at 4 °C overnight with anti-Bax (1: 500, #2774, Cell signaling), anti-Bcl-2 (1: 500, #2876, Proteintech), anti-Aven (1: 1000, #2865, Cell signaling) or anti-GAPDH (1: 500000, 10,494–1-AP, Proteintech).

    Techniques: Inhibition, Flow Cytometry, Quantitation Assay, Expressing, Western Blot

    Bcl-2 overexpression inhibits the activation of the mitochondrial apoptosis pathway by cisplatin in SKOV3 cells. (A and B) Pc-SKOV3 and Bcl-2-SKOV3 cells were treated with 6 µg/ml cisplatin for 6 h and then stained with MitoPotential dye and 7-AAD to assess the Δψm by flow cytometry. (C and D) Western blot analysis of cytochrome c , Bcl-2, Bax, cleaved caspase-9 and cleaved caspase-3 levels in pc-SKOV3 and Bcl-2-SKOV3 cells after cisplatin treatment. The results are the mean ± SD of three independent experiments. *P

    Journal: Oncology Reports

    Article Title: Bcl-2 overexpression reduces cisplatin cytotoxicity by decreasing ER-mitochondrial Ca2+ signaling in SKOV3 cells

    doi: 10.3892/or.2017.6164

    Figure Lengend Snippet: Bcl-2 overexpression inhibits the activation of the mitochondrial apoptosis pathway by cisplatin in SKOV3 cells. (A and B) Pc-SKOV3 and Bcl-2-SKOV3 cells were treated with 6 µg/ml cisplatin for 6 h and then stained with MitoPotential dye and 7-AAD to assess the Δψm by flow cytometry. (C and D) Western blot analysis of cytochrome c , Bcl-2, Bax, cleaved caspase-9 and cleaved caspase-3 levels in pc-SKOV3 and Bcl-2-SKOV3 cells after cisplatin treatment. The results are the mean ± SD of three independent experiments. *P

    Article Snippet: Anti-β-actin (60008–1-Ig; murine Ab; 1:2,000 dilution), anti-Bax (50599–2-Ig; rabbit Ab; 1:2,000 dilution), anti-Bcl-2 (12789–1-AP; rabbit Ab; 1:1,000 dilution), anti-cytochrome c (cyto c ) (10993–1-AP; rabbit Ab; 1:1,000 dilution), anti-Grp78/BIP (11587–1-AP; rabbit Ab; 1:1,000 dilution), peroxidase-conjugated AffiniPure goat anti-mouse IgG (H+L) (SA00001-1; goat Ab; 1:2,000 dilution) and peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H+L) (SA00001-2; goat Ab; 1:2,000 dilution) Abs were purchased from ProteinTech Group, Inc. (Chicago, IL, USA).

    Techniques: Over Expression, Activation Assay, Staining, Flow Cytometry, Cytometry, Western Blot

    Lp MOPV promotes the nuclear translocation of IRF3. (A) HEK 293T cells were cotransfected with Flag-IRF3 and pCMV-Myc (Vec), pCMV-Myc-NP LASV (NP), or pCMV-Myc-Lp MOPV (Lp). At 40 h posttransfection, the cells were collected and then fractionated into nuclear and cytoplasmic fractions. Equal amounts of protein from the cytoplasmic fraction and nuclear fraction were separated by SDS-PAGE and transferred to PVDF membranes. The membranes were then probed with antibodies, and the bands were visualized. Band intensities were measured with Quantity One software. To normalize IRF3 protein intensity, GAPDH and lamin B1 were used as the cytoplasmic and nuclear endogenous loading controls, respectively. (B) HeLa cells were cotransfected with Flag-IRF3 and pCMV-Myc (Vec) or pCMV-Myc-Lp MOPV (Lp). At 36 h posttransfection, the cells were fixed and incubated with the anti-Flag rabbit polyclonal antibody, followed by a DyLight488-labeled antibody to rabbit IgG to detect Flag-IRF3 (green) and the anti-Myc mouse monoclonal antibody followed by a Cy5-labeled antibody to mouse IgG to detect Myc-Lp (red). Next, the cells were stained with DAPI (blue) to visualize nuclei. Cells with Flag-IRF3 signals are indicated by arrows. Immunofluorescence analysis was performed with an A1 MP+ multiphoton confocal microscope (Nikon).

    Journal: Journal of Virology

    Article Title: Activation of the RLR/MAVS Signaling Pathway by the L Protein of Mopeia Virus

    doi: 10.1128/JVI.01292-16

    Figure Lengend Snippet: Lp MOPV promotes the nuclear translocation of IRF3. (A) HEK 293T cells were cotransfected with Flag-IRF3 and pCMV-Myc (Vec), pCMV-Myc-NP LASV (NP), or pCMV-Myc-Lp MOPV (Lp). At 40 h posttransfection, the cells were collected and then fractionated into nuclear and cytoplasmic fractions. Equal amounts of protein from the cytoplasmic fraction and nuclear fraction were separated by SDS-PAGE and transferred to PVDF membranes. The membranes were then probed with antibodies, and the bands were visualized. Band intensities were measured with Quantity One software. To normalize IRF3 protein intensity, GAPDH and lamin B1 were used as the cytoplasmic and nuclear endogenous loading controls, respectively. (B) HeLa cells were cotransfected with Flag-IRF3 and pCMV-Myc (Vec) or pCMV-Myc-Lp MOPV (Lp). At 36 h posttransfection, the cells were fixed and incubated with the anti-Flag rabbit polyclonal antibody, followed by a DyLight488-labeled antibody to rabbit IgG to detect Flag-IRF3 (green) and the anti-Myc mouse monoclonal antibody followed by a Cy5-labeled antibody to mouse IgG to detect Myc-Lp (red). Next, the cells were stained with DAPI (blue) to visualize nuclei. Cells with Flag-IRF3 signals are indicated by arrows. Immunofluorescence analysis was performed with an A1 MP+ multiphoton confocal microscope (Nikon).

    Article Snippet: Mouse monoclonal antibody against the Myc tag (Sigma); mouse monoclonal antibody against double-stranded RNA (dsRNA) (J2; English & Scientific Consulting Kft); rabbit polyclonal antibody against the Myc tag (Cell Signaling Technology); rabbit polyclonal antibodies against glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lamin B1, MAVS, RIG-I, MDA5, and Flag tag (ProteinTech, Wuhan, China); and Cy5-labeled antibody to mouse IgG and DyLight488-labeled antibody to rabbit IgG (KPL) were purchased from the indicated manufacturers.

    Techniques: Translocation Assay, SDS Page, Software, Incubation, Labeling, Staining, Immunofluorescence, Microscopy

    Activation of IFN-I production by Lp MOPV requires MAVS. (A) Knockdown efficiencies of siRNAs against MAVS. HEK 293T cells were transfected with negative-control siRNA (NC) or siRNA against MAVS. At 24 h posttransfection, the cells were collected. Intracellular mRNAs were extracted and subjected to reverse transcription. The relative intracellular mRNA level of MAVS was measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. The intracellular expression level of MAVS protein was analyzed by WB using a polyclonal rabbit antibody for MAVS. (B) Knockdown of MAVS reduces IFN-β promoter activity in SeV-infected cells. HEK 293T cells were transfected with reporter plasmids and the indicated siRNAs and infected with SeV at 24 h posttransfection. At 10 h p.i., the cells were lysed to measure intracellular luciferase activity. (C and D) Knockdown of MAVS reduces both IFN-β promoter activity and intracellular mRNA levels of IFNB1 in Lp MOPV -expressing cells. (C) HEK 293T cells were transfected with the indicated plasmids and siRNAs. At 24 h posttransfection, the cells were collected. To assess IFN-β promoter activity, the cells were lysed to measure intracellular luciferase activity. (D) Intracellular mRNAs were also extracted and subjected to reverse transcription. The relative intracellular mRNA levels of IFNB1 were measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. (E) Overexpression of MAVS activates the IFN-β promoter in Lp MOPV -expressing cells. HEK 293T cells were transfected with the indicated plasmids, and 24 h posttransfection, the cells were lysed to measure intracellular luciferase activity. (F) Knockdown of MAVS does not affect IFN-β promoter activity in cells overexpressing IRF3 and IKK-ε. HEK 293T cells were transfected with the indicated plasmids and siRNAs, and 24 h posttransfection, the cells were lysed to measure intracellular luciferase activity. All experiments were performed at least three times, and the values represent the means and SD of three replicates. *, P

    Journal: Journal of Virology

    Article Title: Activation of the RLR/MAVS Signaling Pathway by the L Protein of Mopeia Virus

    doi: 10.1128/JVI.01292-16

    Figure Lengend Snippet: Activation of IFN-I production by Lp MOPV requires MAVS. (A) Knockdown efficiencies of siRNAs against MAVS. HEK 293T cells were transfected with negative-control siRNA (NC) or siRNA against MAVS. At 24 h posttransfection, the cells were collected. Intracellular mRNAs were extracted and subjected to reverse transcription. The relative intracellular mRNA level of MAVS was measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. The intracellular expression level of MAVS protein was analyzed by WB using a polyclonal rabbit antibody for MAVS. (B) Knockdown of MAVS reduces IFN-β promoter activity in SeV-infected cells. HEK 293T cells were transfected with reporter plasmids and the indicated siRNAs and infected with SeV at 24 h posttransfection. At 10 h p.i., the cells were lysed to measure intracellular luciferase activity. (C and D) Knockdown of MAVS reduces both IFN-β promoter activity and intracellular mRNA levels of IFNB1 in Lp MOPV -expressing cells. (C) HEK 293T cells were transfected with the indicated plasmids and siRNAs. At 24 h posttransfection, the cells were collected. To assess IFN-β promoter activity, the cells were lysed to measure intracellular luciferase activity. (D) Intracellular mRNAs were also extracted and subjected to reverse transcription. The relative intracellular mRNA levels of IFNB1 were measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. (E) Overexpression of MAVS activates the IFN-β promoter in Lp MOPV -expressing cells. HEK 293T cells were transfected with the indicated plasmids, and 24 h posttransfection, the cells were lysed to measure intracellular luciferase activity. (F) Knockdown of MAVS does not affect IFN-β promoter activity in cells overexpressing IRF3 and IKK-ε. HEK 293T cells were transfected with the indicated plasmids and siRNAs, and 24 h posttransfection, the cells were lysed to measure intracellular luciferase activity. All experiments were performed at least three times, and the values represent the means and SD of three replicates. *, P

    Article Snippet: Mouse monoclonal antibody against the Myc tag (Sigma); mouse monoclonal antibody against double-stranded RNA (dsRNA) (J2; English & Scientific Consulting Kft); rabbit polyclonal antibody against the Myc tag (Cell Signaling Technology); rabbit polyclonal antibodies against glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lamin B1, MAVS, RIG-I, MDA5, and Flag tag (ProteinTech, Wuhan, China); and Cy5-labeled antibody to mouse IgG and DyLight488-labeled antibody to rabbit IgG (KPL) were purchased from the indicated manufacturers.

    Techniques: Activation Assay, Transfection, Negative Control, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay, Infection, Luciferase, Over Expression

    Both RIG-I and MDA5 play roles in the Lp MOPV -mediated activation of IFN-I. (A) Knockdown efficiencies of siRNAs against RIG-I. HEK 293T cells were transfected with negative-control siRNA (NC) or siRNA against RIG-I. The cells were collected at 24 h posttransfection. Intracellular mRNAs were extracted and subjected to reverse transcription. The relative intracellular mRNA level of DDX58 was measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. The intracellular expression level of RIG-I protein was analyzed by WB using a polyclonal rabbit antibody for RIG-I. (B) Knockdown of RIG-I reduces IFN-β promoter activity in SeV-infected cells. HEK 293T cells were transfected with reporter plasmids and the indicated siRNAs and infected with SeV at 24 h posttransfection. At 10 h p.i., the cells were lysed to measure intracellular luciferase activity. (C and D) Knockdown of RIG-I reduces both IFN-β promoter activity and intracellular mRNA levels of IFNB1 in Lp MOPV -expressing cells. HEK 293T cells were transfected with the indicated plasmids and siRNAs. The cells were collected at 24 h posttransfection. To assess IFN-β promoter activity, the cells were lysed to measure intracellular luciferase activity. Intracellular mRNAs were also extracted and subjected to reverse transcription. The relative intracellular mRNA levels of IFNB1 were measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. (E) Knockdown efficiencies of siRNAs against MDA5. The knockdown efficiencies of siRNAs against MDA5 were evaluated using the method described for RIG-I. (F) Knockdown of MDA5 reduces IFN-β promoter activity in SeV-infected cells. (G and H) Knockdown of MDA5 reduces both IFN-β promoter activity and intracellular mRNA levels of IFNB1 in Lp MOPV -expressing cells. The effects of the knockdown of MDA5 were evaluated using the method described for RIG-I. All experiments were performed at least three times, and the values represent the means and SD of three replicates. *, P

    Journal: Journal of Virology

    Article Title: Activation of the RLR/MAVS Signaling Pathway by the L Protein of Mopeia Virus

    doi: 10.1128/JVI.01292-16

    Figure Lengend Snippet: Both RIG-I and MDA5 play roles in the Lp MOPV -mediated activation of IFN-I. (A) Knockdown efficiencies of siRNAs against RIG-I. HEK 293T cells were transfected with negative-control siRNA (NC) or siRNA against RIG-I. The cells were collected at 24 h posttransfection. Intracellular mRNAs were extracted and subjected to reverse transcription. The relative intracellular mRNA level of DDX58 was measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. The intracellular expression level of RIG-I protein was analyzed by WB using a polyclonal rabbit antibody for RIG-I. (B) Knockdown of RIG-I reduces IFN-β promoter activity in SeV-infected cells. HEK 293T cells were transfected with reporter plasmids and the indicated siRNAs and infected with SeV at 24 h posttransfection. At 10 h p.i., the cells were lysed to measure intracellular luciferase activity. (C and D) Knockdown of RIG-I reduces both IFN-β promoter activity and intracellular mRNA levels of IFNB1 in Lp MOPV -expressing cells. HEK 293T cells were transfected with the indicated plasmids and siRNAs. The cells were collected at 24 h posttransfection. To assess IFN-β promoter activity, the cells were lysed to measure intracellular luciferase activity. Intracellular mRNAs were also extracted and subjected to reverse transcription. The relative intracellular mRNA levels of IFNB1 were measured by quantitative real-time PCR and calculated using GAPDH as the endogenous control. (E) Knockdown efficiencies of siRNAs against MDA5. The knockdown efficiencies of siRNAs against MDA5 were evaluated using the method described for RIG-I. (F) Knockdown of MDA5 reduces IFN-β promoter activity in SeV-infected cells. (G and H) Knockdown of MDA5 reduces both IFN-β promoter activity and intracellular mRNA levels of IFNB1 in Lp MOPV -expressing cells. The effects of the knockdown of MDA5 were evaluated using the method described for RIG-I. All experiments were performed at least three times, and the values represent the means and SD of three replicates. *, P

    Article Snippet: Mouse monoclonal antibody against the Myc tag (Sigma); mouse monoclonal antibody against double-stranded RNA (dsRNA) (J2; English & Scientific Consulting Kft); rabbit polyclonal antibody against the Myc tag (Cell Signaling Technology); rabbit polyclonal antibodies against glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lamin B1, MAVS, RIG-I, MDA5, and Flag tag (ProteinTech, Wuhan, China); and Cy5-labeled antibody to mouse IgG and DyLight488-labeled antibody to rabbit IgG (KPL) were purchased from the indicated manufacturers.

    Techniques: Activation Assay, Transfection, Negative Control, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay, Infection, Luciferase