mouse anti bax monoclonal  (Millipore)


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    Name:
    Anti Bax antibody
    Description:
    Bax is a 21kD integral organelle membrane protein that belongs to the Bcl 2 family and is expressed mainly in the mitochondria The formation of bax homodimer accelerates apoptotic death whereas its heterodimerization with Bcl 2 or Bcl xL can block apoptosis Monoclonal anti Bax antibody can be used to study the intracellular redistribution of Bax protein upon induction of apoptosis and its unique subcellular localization This product can also be used in immunoblot analysis to estimate variations in the expression of specific proteins involved in apoptosis signalling Mouse anti Bax antibody reacts specifically with the epitope present within the amino acids 12 24 shared by bax from rat mouse and human
    Catalog Number:
    b8429
    Price:
    None
    Applications:
    Anti-Bax antibody, Mouse monoclonal has been used in the western blotting detection in human retinal pigment epithelium cell lines, glioma, and breast cancer cell lines.
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    Structured Review

    Millipore mouse anti bax monoclonal
    Anti Bax antibody
    Bax is a 21kD integral organelle membrane protein that belongs to the Bcl 2 family and is expressed mainly in the mitochondria The formation of bax homodimer accelerates apoptotic death whereas its heterodimerization with Bcl 2 or Bcl xL can block apoptosis Monoclonal anti Bax antibody can be used to study the intracellular redistribution of Bax protein upon induction of apoptosis and its unique subcellular localization This product can also be used in immunoblot analysis to estimate variations in the expression of specific proteins involved in apoptosis signalling Mouse anti Bax antibody reacts specifically with the epitope present within the amino acids 12 24 shared by bax from rat mouse and human
    https://www.bioz.com/result/mouse anti bax monoclonal/product/Millipore
    Average 90 stars, based on 547 article reviews
    Price from $9.99 to $1999.99
    mouse anti bax monoclonal - by Bioz Stars, 2020-02
    90/100 stars

    Images

    1) Product Images from "Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells"

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00320

    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P
    Figure Legend Snippet: Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Techniques Used: Expressing, Cell Culture, Transfection, Western Blot, Positive Control

    Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P
    Figure Legend Snippet: Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Techniques Used: Activation Assay, Expressing, Cell Culture, Transfection, Western Blot

    Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P
    Figure Legend Snippet: Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Techniques Used: Expressing, Western Blot

    2) Product Images from "Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells"

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00320

    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P
    Figure Legend Snippet: Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Techniques Used: Expressing, Cell Culture, Transfection, Western Blot, Positive Control

    Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P
    Figure Legend Snippet: Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Techniques Used: Activation Assay, Expressing, Cell Culture, Transfection, Western Blot

    Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P
    Figure Legend Snippet: Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Techniques Used: Expressing, Western Blot

    3) Product Images from "Development of Oral Epithelial Cell Line ROE2 with Differentiation Potential from Transgenic Rats Harboring Temperature-Sensitive Simian Virus40 Large T-Antigen Gene"

    Article Title: Development of Oral Epithelial Cell Line ROE2 with Differentiation Potential from Transgenic Rats Harboring Temperature-Sensitive Simian Virus40 Large T-Antigen Gene

    Journal: Experimental Animals

    doi: 10.1538/expanim.63.31

    Effects of culture periods and culture temperatures on protein expressions for cytokeratins, P63 and involucrin in ROE2 cells. The cells were cultured at different temperatures for 3, 6, and 9 days. Western blot analyses were performed with specific primary antibodies for Krt4, Krt10, Krt13, Krt14, Krt17/19, P63, involucrin, and Gapdh. Gapdh served as a loading control. Ctr, control (day 0).
    Figure Legend Snippet: Effects of culture periods and culture temperatures on protein expressions for cytokeratins, P63 and involucrin in ROE2 cells. The cells were cultured at different temperatures for 3, 6, and 9 days. Western blot analyses were performed with specific primary antibodies for Krt4, Krt10, Krt13, Krt14, Krt17/19, P63, involucrin, and Gapdh. Gapdh served as a loading control. Ctr, control (day 0).

    Techniques Used: Cell Culture, Western Blot

    Related Articles

    Transduction:

    Article Title: Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
    Article Snippet: For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma). .. For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma).

    Immunocytochemistry:

    Article Title: A neuroprotective agent that inactivates prodegenerative TrkA and preserves mitochondria
    Article Snippet: .. Antibodies The following primary antibodies were used for IHC and WBs: anti–βIII-tubulin (rabbit, IHC, 1:1,000; PRB-435P; Covance), anti–βIII-tubulin (mouse, IHC, 1:1,000; MMS-435P; Covance), anti-caspr, (rabbit, IHC, 1:1,000, provided by E. Peles, Weizmann Institute, Rehovot, Israel), anti–pan Na+ channels (mouse, IHC, 1:1,000; S8809; Sigma-Aldrich); anti-GAPDH (rabbit, WB, 1:1,000; 14C10; Cell Signaling Technology), anti-ERK (rabbit, WB, 1:10,000; K-23; Santa Cruz Biotechnology), anti–phospho-ERK (rabbit, WB, 1:1,000; 9101; Cell Signaling Technology), anti-TrkA (rabbit, WB, 1:500; 2505; Cell Signaling Technology), anti–phospho-TrkA Tyr674/675 (rabbit, WB, 1:500; 4621; Cell Signaling Technology), anti-JNK (rabbit, WB, 1:500; 9251; Cell Signaling Technology), anti-Thr183/Tyr185 phospho-JNK (rabbit, WB, 1:500; 4668; Cell Signaling Technology), anti–cytochrome c (mouse, ICC, 1:550; 612302; BioLegend); anti-MKK4 (rabbit, WB, 1:1,000; 9152; Cell Signaling Technology); anti-Ser257/Thr261 phospho-MKK4 (rabbit, WB, 1:1,000; 9156; Cell Signaling Technology), anti-Ser63 phospho-c-Jun (rabbit, WB, 1:100; 9164; Cell Signaling Technology), anti-Bax (rabbit, WB, 1:500; 2772; Cell Signaling Technology), anti-(active) BAX 6A7 (mouse, WB, 1:100; B8429; Sigma-Aldrich), anti–spectrin αII chain (nonerythroid; mouse, WB, 1:500; MAB1622; EMD Millipore), anti­–α-tubulin (mouse, WB, 1:1,000; T5168; Sigma-Aldrich), anti–cleaved caspase-3 (rabbit, WB, 1:100; IHC, 1:100; AB3623; EMD Millipore), anti-Bim/Bod (rabbit, WB, 1:1,000; ADI-AAP-330-E; Enzo Life Sciences), anti–neurofilament L (rabbit, WB, 1:1,000; 2837; Cell Signaling), anti­–α-internexin (rabbit, 1:1,000; AB5354; Chemicon), anti-SCG10 (rabbit, 1:1,000; 10586-1-AP; Proteintech Group), and anti-Opa1 (rabbit, 1:750; 67589; Cell Signaling Technology). .. The following secondary antibodies (Jackson ImmunoResearch Laboratories) were used: goat anti–rabbit-HRP (1:13,000), goat anti–mouse-HRP (1:10,000), donkey anti–mouse-cy3 or 488 or cy5 (1:1,000), donkey anti–rabbit cy3 or 488 or cy5 (1:1,000), and donkey anti–rat 488 or cy5 (1:500).

    Mass Spectrometry:

    Article Title: Synergistic effects of p53 activation via MDM2 inhibition in combination with inhibition of Bcl-2 or Bcr-Abl in CD34+ proliferating and quiescent chronic myeloid leukemia blast crisis cells
    Article Snippet: .. Antibodies against p53 (#SC-126), MDM2 (#SC-5304), and Mcl-1 (#SC-819) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX); Bax (#554104) from BD Pharmingen (San Jose, CA); Bax (A647) (#B-8429) from Sigma (St. Louis, MO); Puma (ab9643) from Abcam (Cambridge, MS); Bcl-2 (#M0887) from Dako (Carpentaria, CA); and Bcl-xL (#2762s) and p-CrkL (#3181s) from Cell Signaling Technology (Danvers, MA). β-Actin (#A5316) from Sigma was used as a loading control. .. Real-time RT-PCR RT-PCR was carried out as previously described [ , ].

    MTT Assay:

    Article Title: Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species
    Article Snippet: .. MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazoliumbromide), GSH, 5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB), thiobarbituric acid (TBA), 2,7-dichlorofluorescin diacetate (DCFH-DA), anti-p53 antibody, anti-bax antibody, anti-bcl-2 antibody, and anti-β-actin antibody were obtained from Sigma-Aldrich (St Louis, MO). .. Secondary antibodies, RIPA buffer, and sodium dodecyl sulfate (SDS) were bought from Santa Cruz Biotechnology, Inc, (Santa Cruz, CA).

    Cytometry:

    Article Title: Dynein light chain 1 induces assembly of large Bim complexes on mitochondria that stabilize Mcl-1 and regulate apoptosis
    Article Snippet: .. For flow cytometry experiments, treated cells were fixed in 4% formalin for 15 min and stained with conformation-specific primary antibodies (Bax 6A7 [Sigma, no. B8429] and Bak NT [Millipore, no. 06536]). .. Species-specific Alexa fluor 647 or Cy5-conjugated secondary antibodies were used to detect activated Bax and Bak by flow cytometry.

    Blocking Assay:

    Article Title: Two venom allergen‐like proteins, HaVAP1 and HaVAP2, are involved in the parasitism of Heterodera avenae
    Article Snippet: .. After blocking, the membrane was incubated with an anti‐BAX antibody (Sigma‐Aldrich) to bind BAX. .. Horseradish peroxidase (HRP)‐conjugated goat anti‐mouse immunoglobulin G (IgG) (Sigma) was used as the secondary antibody.

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: .. Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China). .. A chemiluminescence detection kit (Beyotime Institute of Biotechnology) was employed to detect the labeled bands. β-actin (1:1,000; Sangon Biotech Co., Ltd., Shanghai, China) served as an internal reference and the relative protein expression of target proteins was determined.

    Article Title: Safflor yellow B reduces hypoxia-mediated vasoconstriction by regulating endothelial micro ribonucleic acid/nitric oxide synthase signaling
    Article Snippet: .. After blocking with 1% bovine serum albumin for 120 min at 20 ± 2°C, the membranes were incubated with one of the following primary antibodies: anti-HIF-1α (1:1000, ab216842), anti-caspase 3 (1:500, ab32042), anti-eNOS (1:500, ab95254), anti-p-eNOSSer1177 (1:500, ab51038, all Abcam), anti-Bcl-2 (1:1000, SAB4500003), anti-Bax (1:1000, SAB4502546), anti-iNOS (1:1000, SAB4502011), anti-p-iNOSTyr151 (1:1000, SAB4301563), and anti-S100A1 (1:250, SAB4502708, all Sigma-Aldrich) overnight at 4°C prior to washing thrice (15-min each) with Tris-buffered saline containing Tween 20 (TBST). .. The blots were then incubated with the corresponding HRP-conjugated secondary antibody for 50–60 min at 20 ± 2°C, followed by three 10-min washes with TBST.

    Electrophoresis:

    Article Title: Safflor yellow B reduces hypoxia-mediated vasoconstriction by regulating endothelial micro ribonucleic acid/nitric oxide synthase signaling
    Article Snippet: Equal amounts of total proteins (40 μg) were separated using sodium dodecyl sulfate-polyacrylamide gel (15%) electrophoresis and then transferred to a polyvinylidene difluoride (PVDF) membrane (IPVH00010, Millipore, Bedford, MA, USA). .. After blocking with 1% bovine serum albumin for 120 min at 20 ± 2°C, the membranes were incubated with one of the following primary antibodies: anti-HIF-1α (1:1000, ab216842), anti-caspase 3 (1:500, ab32042), anti-eNOS (1:500, ab95254), anti-p-eNOSSer1177 (1:500, ab51038, all Abcam), anti-Bcl-2 (1:1000, SAB4500003), anti-Bax (1:1000, SAB4502546), anti-iNOS (1:1000, SAB4502011), anti-p-iNOSTyr151 (1:1000, SAB4301563), and anti-S100A1 (1:250, SAB4502708, all Sigma-Aldrich) overnight at 4°C prior to washing thrice (15-min each) with Tris-buffered saline containing Tween 20 (TBST).

    Incubation:

    Article Title: Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
    Article Snippet: .. For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma). .. Beads were washed 4 times with 1 X IP50 buffer and twice with 0.1 X IP50 buffer, and incubated with 20 µL Laemmli buffer, before SDS-PAGE and Western-blotting.

    Article Title: Two venom allergen‐like proteins, HaVAP1 and HaVAP2, are involved in the parasitism of Heterodera avenae
    Article Snippet: .. After blocking, the membrane was incubated with an anti‐BAX antibody (Sigma‐Aldrich) to bind BAX. .. Horseradish peroxidase (HRP)‐conjugated goat anti‐mouse immunoglobulin G (IgG) (Sigma) was used as the secondary antibody.

    Article Title: Dynein light chain 1 induces assembly of large Bim complexes on mitochondria that stabilize Mcl-1 and regulate apoptosis
    Article Snippet: After incubation, the reaction was stopped by adding 1 mM DTT for an additional 15 min at room temperature. .. For flow cytometry experiments, treated cells were fixed in 4% formalin for 15 min and stained with conformation-specific primary antibodies (Bax 6A7 [Sigma, no. B8429] and Bak NT [Millipore, no. 06536]).

    Article Title: Targeting redox homeostasis in rhabdomyosarcoma cells: GSH-depleting agents enhance auranofin-induced cell death
    Article Snippet: .. 500–1000 μ g protein was immunoprecipitated and incubated overnight at 4 °C with 2 μ g/ml mouse anti-BAK antibody (Ab-1; Merck Millipore, Billerica, MA, USA) or anti-BAX antibody (6A7, Sigma-Aldrich) and 10 μ l pan-mouse IgG Dynabeads (Life technologies, Inc.), washed with CHAPS lysis buffer and analyzed by western blotting using rabbit anti-BAK antibody (BD Bioscience) or anti-BAX antibody (Merck, Darmstadt, Germany). .. Immunoprecipitation of MCL-1 was performed in 500 μ l lysates containing up to 1000 μ g proteins, which were incubated overnight at 4 °C with 2 μ g/ml mouse anti-MCL-1 antibody (BD Biosciences) and 10 μ l pan-mouse IgG Dynabeads or Protein G Dynabeads (Life Technologies, Inc.) and washed with CHAPS buffer.

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: .. Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China). .. A chemiluminescence detection kit (Beyotime Institute of Biotechnology) was employed to detect the labeled bands. β-actin (1:1,000; Sangon Biotech Co., Ltd., Shanghai, China) served as an internal reference and the relative protein expression of target proteins was determined.

    Article Title: Safflor yellow B reduces hypoxia-mediated vasoconstriction by regulating endothelial micro ribonucleic acid/nitric oxide synthase signaling
    Article Snippet: .. After blocking with 1% bovine serum albumin for 120 min at 20 ± 2°C, the membranes were incubated with one of the following primary antibodies: anti-HIF-1α (1:1000, ab216842), anti-caspase 3 (1:500, ab32042), anti-eNOS (1:500, ab95254), anti-p-eNOSSer1177 (1:500, ab51038, all Abcam), anti-Bcl-2 (1:1000, SAB4500003), anti-Bax (1:1000, SAB4502546), anti-iNOS (1:1000, SAB4502011), anti-p-iNOSTyr151 (1:1000, SAB4301563), and anti-S100A1 (1:250, SAB4502708, all Sigma-Aldrich) overnight at 4°C prior to washing thrice (15-min each) with Tris-buffered saline containing Tween 20 (TBST). .. The blots were then incubated with the corresponding HRP-conjugated secondary antibody for 50–60 min at 20 ± 2°C, followed by three 10-min washes with TBST.

    Expressing:

    Article Title: Two venom allergen‐like proteins, HaVAP1 and HaVAP2, are involved in the parasitism of Heterodera avenae
    Article Snippet: Western blotting was employed to confirm the expression of BAX. .. After blocking, the membrane was incubated with an anti‐BAX antibody (Sigma‐Aldrich) to bind BAX.

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China). .. A chemiluminescence detection kit (Beyotime Institute of Biotechnology) was employed to detect the labeled bands. β-actin (1:1,000; Sangon Biotech Co., Ltd., Shanghai, China) served as an internal reference and the relative protein expression of target proteins was determined.

    BIA-KA:

    Article Title: Rapid induction of apoptosis during Kinesin-5 inhibitor-induced mitotic arrest in HL60 cells
    Article Snippet: Supernatant was collected and protein concentration was determined using Pierce BCA Protein Assay Kit. .. Immunoprecipitation of activated Bax using anti-Bax antibody clone 6A7 (Sigma) was described previously [ ].

    Western Blot:

    Article Title: Synergistic effects of p53 activation via MDM2 inhibition in combination with inhibition of Bcl-2 or Bcr-Abl in CD34+ proliferating and quiescent chronic myeloid leukemia blast crisis cells
    Article Snippet: Paragraph title: Western blot ... Antibodies against p53 (#SC-126), MDM2 (#SC-5304), and Mcl-1 (#SC-819) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX); Bax (#554104) from BD Pharmingen (San Jose, CA); Bax (A647) (#B-8429) from Sigma (St. Louis, MO); Puma (ab9643) from Abcam (Cambridge, MS); Bcl-2 (#M0887) from Dako (Carpentaria, CA); and Bcl-xL (#2762s) and p-CrkL (#3181s) from Cell Signaling Technology (Danvers, MA). β-Actin (#A5316) from Sigma was used as a loading control.

    Article Title: Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
    Article Snippet: For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma). .. Beads were washed 4 times with 1 X IP50 buffer and twice with 0.1 X IP50 buffer, and incubated with 20 µL Laemmli buffer, before SDS-PAGE and Western-blotting.

    Article Title: Rapid induction of apoptosis during Kinesin-5 inhibitor-induced mitotic arrest in HL60 cells
    Article Snippet: Paragraph title: 2.6 Western Blot and Immunoprecipitation ... Immunoprecipitation of activated Bax using anti-Bax antibody clone 6A7 (Sigma) was described previously [ ].

    Article Title: Two venom allergen‐like proteins, HaVAP1 and HaVAP2, are involved in the parasitism of Heterodera avenae
    Article Snippet: Western blotting was employed to confirm the expression of BAX. .. After blocking, the membrane was incubated with an anti‐BAX antibody (Sigma‐Aldrich) to bind BAX.

    Article Title: Targeting redox homeostasis in rhabdomyosarcoma cells: GSH-depleting agents enhance auranofin-induced cell death
    Article Snippet: .. 500–1000 μ g protein was immunoprecipitated and incubated overnight at 4 °C with 2 μ g/ml mouse anti-BAK antibody (Ab-1; Merck Millipore, Billerica, MA, USA) or anti-BAX antibody (6A7, Sigma-Aldrich) and 10 μ l pan-mouse IgG Dynabeads (Life technologies, Inc.), washed with CHAPS lysis buffer and analyzed by western blotting using rabbit anti-BAK antibody (BD Bioscience) or anti-BAX antibody (Merck, Darmstadt, Germany). .. Immunoprecipitation of MCL-1 was performed in 500 μ l lysates containing up to 1000 μ g proteins, which were incubated overnight at 4 °C with 2 μ g/ml mouse anti-MCL-1 antibody (BD Biosciences) and 10 μ l pan-mouse IgG Dynabeads or Protein G Dynabeads (Life Technologies, Inc.) and washed with CHAPS buffer.

    Article Title: A neuroprotective agent that inactivates prodegenerative TrkA and preserves mitochondria
    Article Snippet: .. Antibodies The following primary antibodies were used for IHC and WBs: anti–βIII-tubulin (rabbit, IHC, 1:1,000; PRB-435P; Covance), anti–βIII-tubulin (mouse, IHC, 1:1,000; MMS-435P; Covance), anti-caspr, (rabbit, IHC, 1:1,000, provided by E. Peles, Weizmann Institute, Rehovot, Israel), anti–pan Na+ channels (mouse, IHC, 1:1,000; S8809; Sigma-Aldrich); anti-GAPDH (rabbit, WB, 1:1,000; 14C10; Cell Signaling Technology), anti-ERK (rabbit, WB, 1:10,000; K-23; Santa Cruz Biotechnology), anti–phospho-ERK (rabbit, WB, 1:1,000; 9101; Cell Signaling Technology), anti-TrkA (rabbit, WB, 1:500; 2505; Cell Signaling Technology), anti–phospho-TrkA Tyr674/675 (rabbit, WB, 1:500; 4621; Cell Signaling Technology), anti-JNK (rabbit, WB, 1:500; 9251; Cell Signaling Technology), anti-Thr183/Tyr185 phospho-JNK (rabbit, WB, 1:500; 4668; Cell Signaling Technology), anti–cytochrome c (mouse, ICC, 1:550; 612302; BioLegend); anti-MKK4 (rabbit, WB, 1:1,000; 9152; Cell Signaling Technology); anti-Ser257/Thr261 phospho-MKK4 (rabbit, WB, 1:1,000; 9156; Cell Signaling Technology), anti-Ser63 phospho-c-Jun (rabbit, WB, 1:100; 9164; Cell Signaling Technology), anti-Bax (rabbit, WB, 1:500; 2772; Cell Signaling Technology), anti-(active) BAX 6A7 (mouse, WB, 1:100; B8429; Sigma-Aldrich), anti–spectrin αII chain (nonerythroid; mouse, WB, 1:500; MAB1622; EMD Millipore), anti­–α-tubulin (mouse, WB, 1:1,000; T5168; Sigma-Aldrich), anti–cleaved caspase-3 (rabbit, WB, 1:100; IHC, 1:100; AB3623; EMD Millipore), anti-Bim/Bod (rabbit, WB, 1:1,000; ADI-AAP-330-E; Enzo Life Sciences), anti–neurofilament L (rabbit, WB, 1:1,000; 2837; Cell Signaling), anti­–α-internexin (rabbit, 1:1,000; AB5354; Chemicon), anti-SCG10 (rabbit, 1:1,000; 10586-1-AP; Proteintech Group), and anti-Opa1 (rabbit, 1:750; 67589; Cell Signaling Technology). .. The following secondary antibodies (Jackson ImmunoResearch Laboratories) were used: goat anti–rabbit-HRP (1:13,000), goat anti–mouse-HRP (1:10,000), donkey anti–mouse-cy3 or 488 or cy5 (1:1,000), donkey anti–rabbit cy3 or 488 or cy5 (1:1,000), and donkey anti–rat 488 or cy5 (1:500).

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Paragraph title: Western blot analysis ... Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China).

    Article Title: Recombinant VP1, an Akt Inhibitor, Suppresses Progression of Hepatocellular Carcinoma by Inducing Apoptosis and Modulation of CCL2 Production
    Article Snippet: Immunoprecipitation For detection of Ku70-Bax interactions in BNL and Hepa1-6 cells, cells treated with or without rVP1 were harvested in protein extraction reagents (Pierce), and 0.2–1 mg of cell lysates were immunoprecipitated with 4–10 µg of anti-Bax antibody (Sigma). .. After washing with PBS five times, western blot analysis of pre-immunoprecipitated (Input) and immunoprecipitated (IP) samples was performed with an anti-Ku70 antibody.

    Article Title: Safflor yellow B reduces hypoxia-mediated vasoconstriction by regulating endothelial micro ribonucleic acid/nitric oxide synthase signaling
    Article Snippet: Paragraph title: Western blot analysis ... After blocking with 1% bovine serum albumin for 120 min at 20 ± 2°C, the membranes were incubated with one of the following primary antibodies: anti-HIF-1α (1:1000, ab216842), anti-caspase 3 (1:500, ab32042), anti-eNOS (1:500, ab95254), anti-p-eNOSSer1177 (1:500, ab51038, all Abcam), anti-Bcl-2 (1:1000, SAB4500003), anti-Bax (1:1000, SAB4502546), anti-iNOS (1:1000, SAB4502011), anti-p-iNOSTyr151 (1:1000, SAB4301563), and anti-S100A1 (1:250, SAB4502708, all Sigma-Aldrich) overnight at 4°C prior to washing thrice (15-min each) with Tris-buffered saline containing Tween 20 (TBST).

    Flow Cytometry:

    Article Title: Dynein light chain 1 induces assembly of large Bim complexes on mitochondria that stabilize Mcl-1 and regulate apoptosis
    Article Snippet: .. For flow cytometry experiments, treated cells were fixed in 4% formalin for 15 min and stained with conformation-specific primary antibodies (Bax 6A7 [Sigma, no. B8429] and Bak NT [Millipore, no. 06536]). .. Species-specific Alexa fluor 647 or Cy5-conjugated secondary antibodies were used to detect activated Bax and Bak by flow cytometry.

    Transfection:

    Article Title: Farnesylthiosalicylic acid sensitizes hepatocarcinoma cells to artemisinin derivatives
    Article Snippet: Mouse monoclonal anti-Bak antibody (Ab-2) and anti-Bax antibody (6A7) were purchased from Calbiochem (San Diego, CA, USA). .. Turbofect™ transfection regent was purchased from Thermo Fisher Scientific (Massachusetts, USA).

    Immunoprecipitation:

    Article Title: Microtubule-Targeting Drugs Induce Bcl-2 Phosphorylation and Association with Pin1
    Article Snippet: .. After normalization for either input cell number or total protein content, the resulting lysates were subjected to immunoprecipitation with rabbit antipeptide anti-Bcl-2 or anti-Bax antibodies using Protein A Sepharose (from Sigma) or with anti-Bcl-2 monoclonals using Protein G Sepharose (Zymed, San Francisco, CA). ..

    Article Title: Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
    Article Snippet: .. For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma). .. Beads were washed 4 times with 1 X IP50 buffer and twice with 0.1 X IP50 buffer, and incubated with 20 µL Laemmli buffer, before SDS-PAGE and Western-blotting.

    Article Title: Rapid induction of apoptosis during Kinesin-5 inhibitor-induced mitotic arrest in HL60 cells
    Article Snippet: .. Immunoprecipitation of activated Bax using anti-Bax antibody clone 6A7 (Sigma) was described previously [ ]. .. Cell pellets were lysed in 1% CHAPS lysis buffer (150mM NaCl, 10mM HEPES (pH 7.4), 1% CHAPS) with a cocktail of protease inhibitors on ice for 30 minutes.

    Article Title: Targeting redox homeostasis in rhabdomyosarcoma cells: GSH-depleting agents enhance auranofin-induced cell death
    Article Snippet: .. 500–1000 μ g protein was immunoprecipitated and incubated overnight at 4 °C with 2 μ g/ml mouse anti-BAK antibody (Ab-1; Merck Millipore, Billerica, MA, USA) or anti-BAX antibody (6A7, Sigma-Aldrich) and 10 μ l pan-mouse IgG Dynabeads (Life technologies, Inc.), washed with CHAPS lysis buffer and analyzed by western blotting using rabbit anti-BAK antibody (BD Bioscience) or anti-BAX antibody (Merck, Darmstadt, Germany). .. Immunoprecipitation of MCL-1 was performed in 500 μ l lysates containing up to 1000 μ g proteins, which were incubated overnight at 4 °C with 2 μ g/ml mouse anti-MCL-1 antibody (BD Biosciences) and 10 μ l pan-mouse IgG Dynabeads or Protein G Dynabeads (Life Technologies, Inc.) and washed with CHAPS buffer.

    Article Title: Recombinant VP1, an Akt Inhibitor, Suppresses Progression of Hepatocellular Carcinoma by Inducing Apoptosis and Modulation of CCL2 Production
    Article Snippet: .. Immunoprecipitation For detection of Ku70-Bax interactions in BNL and Hepa1-6 cells, cells treated with or without rVP1 were harvested in protein extraction reagents (Pierce), and 0.2–1 mg of cell lysates were immunoprecipitated with 4–10 µg of anti-Bax antibody (Sigma). .. The immunocomplexes were captured using an immunoprecipitation matrix (ExactaCruz C, Santa Cruz Biotechnology) following the manufacturer's protocol.

    Protease Inhibitor:

    Article Title: Rapid induction of apoptosis during Kinesin-5 inhibitor-induced mitotic arrest in HL60 cells
    Article Snippet: Harvested cell pellets were lysed in lysis buffer (50mM Tris pH 7.5, 100mM NaCl, 1% Triton X-100, 0.5% deoxycholate, 0.1% SDS, 1x protease inhibitor cocktail, 50mM NaF, phosphatase inhibitor cocktail) for 30min on ice. .. Immunoprecipitation of activated Bax using anti-Bax antibody clone 6A7 (Sigma) was described previously [ ].

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Cell lysates were prepared from the cell sample using radioimmunoprecipitation assay lysis buffer (Beyotime Institute of Biotechnology) combined with a protease inhibitor. .. Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China).

    other:

    Article Title: Proapoptotic Protein Smac Mediates Apoptosis in Cisplatin-resistant Ovarian Cancer Cells When Treated with the Anti-tumor Agent AT101 *
    Article Snippet: PI and Bax (clone 6A7, B8429), FLAG (clone M1, F3040), and actin (clone AC-74, A5316) antibodies were obtained from Sigma.

    Protein Concentration:

    Article Title: Rapid induction of apoptosis during Kinesin-5 inhibitor-induced mitotic arrest in HL60 cells
    Article Snippet: Supernatant was collected and protein concentration was determined using Pierce BCA Protein Assay Kit. .. Immunoprecipitation of activated Bax using anti-Bax antibody clone 6A7 (Sigma) was described previously [ ].

    Nucleic Acid Electrophoresis:

    Article Title: Two venom allergen‐like proteins, HaVAP1 and HaVAP2, are involved in the parasitism of Heterodera avenae
    Article Snippet: Protein crude extracts were separated by sodium dodecylsulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane. .. After blocking, the membrane was incubated with an anti‐BAX antibody (Sigma‐Aldrich) to bind BAX.

    Isolation:

    Article Title: Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
    Article Snippet: Most experiments have been done immediately on freshly isolated fractions, but some additional experiments have been done on mitochondrial suspension frozen as small beads in liquid nitrogen. .. For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma).

    Article Title: Dynein light chain 1 induces assembly of large Bim complexes on mitochondria that stabilize Mcl-1 and regulate apoptosis
    Article Snippet: The isolated mitochondria (30–60 µg) were incubated for 30 min in the dark at room temperature in 45 µL of cross-linking buffer (20 mM HEPES at pH 7.5, 250 mM sucrose, 1 mM EDTA, 50 mM KCl, 2.5 mM MgCl2 ) containing 0.5 mM BMH (1, 6–bismaleimidohexane; Thermo Scientific, no. 22331). .. For flow cytometry experiments, treated cells were fixed in 4% formalin for 15 min and stained with conformation-specific primary antibodies (Bax 6A7 [Sigma, no. B8429] and Bak NT [Millipore, no. 06536]).

    Bicinchoninic Acid Protein Assay:

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Protein concentrations were determined using a bicinchoninic acid protein assay kit (Thermo Fisher Scientific, Inc.). .. Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China).

    Immunohistochemistry:

    Article Title: A neuroprotective agent that inactivates prodegenerative TrkA and preserves mitochondria
    Article Snippet: .. Antibodies The following primary antibodies were used for IHC and WBs: anti–βIII-tubulin (rabbit, IHC, 1:1,000; PRB-435P; Covance), anti–βIII-tubulin (mouse, IHC, 1:1,000; MMS-435P; Covance), anti-caspr, (rabbit, IHC, 1:1,000, provided by E. Peles, Weizmann Institute, Rehovot, Israel), anti–pan Na+ channels (mouse, IHC, 1:1,000; S8809; Sigma-Aldrich); anti-GAPDH (rabbit, WB, 1:1,000; 14C10; Cell Signaling Technology), anti-ERK (rabbit, WB, 1:10,000; K-23; Santa Cruz Biotechnology), anti–phospho-ERK (rabbit, WB, 1:1,000; 9101; Cell Signaling Technology), anti-TrkA (rabbit, WB, 1:500; 2505; Cell Signaling Technology), anti–phospho-TrkA Tyr674/675 (rabbit, WB, 1:500; 4621; Cell Signaling Technology), anti-JNK (rabbit, WB, 1:500; 9251; Cell Signaling Technology), anti-Thr183/Tyr185 phospho-JNK (rabbit, WB, 1:500; 4668; Cell Signaling Technology), anti–cytochrome c (mouse, ICC, 1:550; 612302; BioLegend); anti-MKK4 (rabbit, WB, 1:1,000; 9152; Cell Signaling Technology); anti-Ser257/Thr261 phospho-MKK4 (rabbit, WB, 1:1,000; 9156; Cell Signaling Technology), anti-Ser63 phospho-c-Jun (rabbit, WB, 1:100; 9164; Cell Signaling Technology), anti-Bax (rabbit, WB, 1:500; 2772; Cell Signaling Technology), anti-(active) BAX 6A7 (mouse, WB, 1:100; B8429; Sigma-Aldrich), anti–spectrin αII chain (nonerythroid; mouse, WB, 1:500; MAB1622; EMD Millipore), anti­–α-tubulin (mouse, WB, 1:1,000; T5168; Sigma-Aldrich), anti–cleaved caspase-3 (rabbit, WB, 1:100; IHC, 1:100; AB3623; EMD Millipore), anti-Bim/Bod (rabbit, WB, 1:1,000; ADI-AAP-330-E; Enzo Life Sciences), anti–neurofilament L (rabbit, WB, 1:1,000; 2837; Cell Signaling), anti­–α-internexin (rabbit, 1:1,000; AB5354; Chemicon), anti-SCG10 (rabbit, 1:1,000; 10586-1-AP; Proteintech Group), and anti-Opa1 (rabbit, 1:750; 67589; Cell Signaling Technology). .. The following secondary antibodies (Jackson ImmunoResearch Laboratories) were used: goat anti–rabbit-HRP (1:13,000), goat anti–mouse-HRP (1:10,000), donkey anti–mouse-cy3 or 488 or cy5 (1:1,000), donkey anti–rabbit cy3 or 488 or cy5 (1:1,000), and donkey anti–rat 488 or cy5 (1:500).

    Article Title: Deguelin inhibits non-small cell lung cancer via down-regulating Hexokinases II-mediated glycolysis
    Article Snippet: Antibodies against HK2 (2867, IB: 1:2000, IHC:1:200), p-Akt (4060, IB: 1:1000, IHC: 1:100), Akt (4691, IB: 1:2000), Akt1 (75692, IB:1:2000), p-S6 (4858, IB: 1:4000), S6 (2317, IB:1:2000), VDAC1 (4866, IB:1:2000), cleaved-caspase 3 (9664, IB: 1:2000), cleaved-caspase 9 (9505, IB: 1:1000), cleaved-PARP (5625, IB: 1:2000) and cytochrome C (4280, IB: 1:1000) were obtained from Cell Signaling Technology, Inc. (Beverly, MA). .. Antibody against β-actin (A5316, IB: 1:10000) and Bax (B8429, IB: 1:2000) were from Sigma-Aldrich.

    Labeling:

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China). .. A chemiluminescence detection kit (Beyotime Institute of Biotechnology) was employed to detect the labeled bands. β-actin (1:1,000; Sangon Biotech Co., Ltd., Shanghai, China) served as an internal reference and the relative protein expression of target proteins was determined.

    Immunodetection:

    Article Title: Microtubule-Targeting Drugs Induce Bcl-2 Phosphorylation and Association with Pin1
    Article Snippet: After normalization for either input cell number or total protein content, the resulting lysates were subjected to immunoprecipitation with rabbit antipeptide anti-Bcl-2 or anti-Bax antibodies using Protein A Sepharose (from Sigma) or with anti-Bcl-2 monoclonals using Protein G Sepharose (Zymed, San Francisco, CA). .. Immunodetection was accomplished by an enhanced chemiluminescence (ECL) method (Amersham/Pharmacia, Piscataway, NJ).

    Protein Extraction:

    Article Title: Two venom allergen‐like proteins, HaVAP1 and HaVAP2, are involved in the parasitism of Heterodera avenae
    Article Snippet: Total proteins of infiltrated leaves were extracted using a Plant Protein Extraction Kit (CWBio, Beijing, China). .. After blocking, the membrane was incubated with an anti‐BAX antibody (Sigma‐Aldrich) to bind BAX.

    Article Title: Recombinant VP1, an Akt Inhibitor, Suppresses Progression of Hepatocellular Carcinoma by Inducing Apoptosis and Modulation of CCL2 Production
    Article Snippet: .. Immunoprecipitation For detection of Ku70-Bax interactions in BNL and Hepa1-6 cells, cells treated with or without rVP1 were harvested in protein extraction reagents (Pierce), and 0.2–1 mg of cell lysates were immunoprecipitated with 4–10 µg of anti-Bax antibody (Sigma). .. The immunocomplexes were captured using an immunoprecipitation matrix (ExactaCruz C, Santa Cruz Biotechnology) following the manufacturer's protocol.

    Lysis:

    Article Title: Microtubule-Targeting Drugs Induce Bcl-2 Phosphorylation and Association with Pin1
    Article Snippet: Cells were lysed in ice-cold RIPA buffer (10 mM Tris pH 7.4, 142.5 mM NaCl, 1% Na deoxycholate, 0.1% SDS, 1% Triton X-100, 1 mM EDTA) or NP-40 lysis buffer (10 mM Tris pH 7.4, 142.5 mM KCl, 0.5% NP-40, 5 mM EGTA), containing protease inhibitors and phosphatase inhibitors (10 mM sodium β -glycerophosphate, 1 mM Na3 VO4 , 5 mM NaF, 2 mM Na4 P2 O7 , 50 mM and 4-nitrophenyl phosphate). .. After normalization for either input cell number or total protein content, the resulting lysates were subjected to immunoprecipitation with rabbit antipeptide anti-Bcl-2 or anti-Bax antibodies using Protein A Sepharose (from Sigma) or with anti-Bcl-2 monoclonals using Protein G Sepharose (Zymed, San Francisco, CA).

    Article Title: Rapid induction of apoptosis during Kinesin-5 inhibitor-induced mitotic arrest in HL60 cells
    Article Snippet: Harvested cell pellets were lysed in lysis buffer (50mM Tris pH 7.5, 100mM NaCl, 1% Triton X-100, 0.5% deoxycholate, 0.1% SDS, 1x protease inhibitor cocktail, 50mM NaF, phosphatase inhibitor cocktail) for 30min on ice. .. Immunoprecipitation of activated Bax using anti-Bax antibody clone 6A7 (Sigma) was described previously [ ].

    Article Title: Targeting redox homeostasis in rhabdomyosarcoma cells: GSH-depleting agents enhance auranofin-induced cell death
    Article Snippet: .. 500–1000 μ g protein was immunoprecipitated and incubated overnight at 4 °C with 2 μ g/ml mouse anti-BAK antibody (Ab-1; Merck Millipore, Billerica, MA, USA) or anti-BAX antibody (6A7, Sigma-Aldrich) and 10 μ l pan-mouse IgG Dynabeads (Life technologies, Inc.), washed with CHAPS lysis buffer and analyzed by western blotting using rabbit anti-BAK antibody (BD Bioscience) or anti-BAX antibody (Merck, Darmstadt, Germany). .. Immunoprecipitation of MCL-1 was performed in 500 μ l lysates containing up to 1000 μ g proteins, which were incubated overnight at 4 °C with 2 μ g/ml mouse anti-MCL-1 antibody (BD Biosciences) and 10 μ l pan-mouse IgG Dynabeads or Protein G Dynabeads (Life Technologies, Inc.) and washed with CHAPS buffer.

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Cell lysates were prepared from the cell sample using radioimmunoprecipitation assay lysis buffer (Beyotime Institute of Biotechnology) combined with a protease inhibitor. .. Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China).

    Article Title: Safflor yellow B reduces hypoxia-mediated vasoconstriction by regulating endothelial micro ribonucleic acid/nitric oxide synthase signaling
    Article Snippet: Vascular endothelial cell or RAEC samples were homogenized with lysis buffer and centrifuged at 12,000 g for 15 min. .. After blocking with 1% bovine serum albumin for 120 min at 20 ± 2°C, the membranes were incubated with one of the following primary antibodies: anti-HIF-1α (1:1000, ab216842), anti-caspase 3 (1:500, ab32042), anti-eNOS (1:500, ab95254), anti-p-eNOSSer1177 (1:500, ab51038, all Abcam), anti-Bcl-2 (1:1000, SAB4500003), anti-Bax (1:1000, SAB4502546), anti-iNOS (1:1000, SAB4502011), anti-p-iNOSTyr151 (1:1000, SAB4301563), and anti-S100A1 (1:250, SAB4502708, all Sigma-Aldrich) overnight at 4°C prior to washing thrice (15-min each) with Tris-buffered saline containing Tween 20 (TBST).

    SDS Page:

    Article Title: Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
    Article Snippet: 100 µg of proteins were separated by SDS-PAGE (12.5% acrylamide), blotted on PVDF, and revealed with antibodies against Bax, Bcl-xL, Porin and PGK (as mitochondrial and cytosolic markers, respectively). .. For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma).

    Article Title: Two venom allergen‐like proteins, HaVAP1 and HaVAP2, are involved in the parasitism of Heterodera avenae
    Article Snippet: Protein crude extracts were separated by sodium dodecylsulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane. .. After blocking, the membrane was incubated with an anti‐BAX antibody (Sigma‐Aldrich) to bind BAX.

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Proteins from each sample (40 µg) were separated by 10% SDS-PAGE, and transferred to polyvinylidene fluoride membranes (Merck KGaA, Darmstadt, Germany). .. Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China).

    Negative Control:

    Article Title: Recombinant VP1, an Akt Inhibitor, Suppresses Progression of Hepatocellular Carcinoma by Inducing Apoptosis and Modulation of CCL2 Production
    Article Snippet: Immunoprecipitation For detection of Ku70-Bax interactions in BNL and Hepa1-6 cells, cells treated with or without rVP1 were harvested in protein extraction reagents (Pierce), and 0.2–1 mg of cell lysates were immunoprecipitated with 4–10 µg of anti-Bax antibody (Sigma). .. Mouse IgG (Zymed, San Francisco, CA) was used as a negative control.

    Radio Immunoprecipitation:

    Article Title: Different inhibitory effects on the proliferation and apoptosis of human and laboratory Borna disease virus-infected human neuroblastoma SH-SY5Y cells in vitro
    Article Snippet: Cell lysates were prepared from the cell sample using radioimmunoprecipitation assay lysis buffer (Beyotime Institute of Biotechnology) combined with a protease inhibitor. .. Following blocking by 5% fetal bovine serum for 12 h at 4°C, the membranes were probed with the primary antibodies rabbit monoclonal anti-apoptosis regulator BAX (Bax; Sigma-Aldrich; Merck KGaA; cat. no. SAB5500012) and anti-apoptosis regulator Bcl-2 (Bcl-2; 1:1,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody for 1 h at 25°C (1:1,000; cat. no. AB501-01A; NovoProtein Biotech Co., Ltd., Shanghai, China).

    CCK-8 Assay:

    Article Title: Farnesylthiosalicylic acid sensitizes hepatocarcinoma cells to artemisinin derivatives
    Article Snippet: Mouse monoclonal anti-Bak antibody (Ab-2) and anti-Bax antibody (6A7) were purchased from Calbiochem (San Diego, CA, USA). .. Mouse monoclonal anti-Bak antibody (Ab-2) and anti-Bax antibody (6A7) were purchased from Calbiochem (San Diego, CA, USA).

    Spectrophotometry:

    Article Title: Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
    Article Snippet: For immunoprecipitation, 2 mg proteins from mitochondria and S30 fractions were solubilized for 30 minutes in 0.5 mL of 1 X IP50 buffer from Sigma, then incubated overnight with 2 µg anti-Bax antibody (2D2, Sigma), and 4 additional hours with protein 50 µL G-sepharose beads (Sigma). .. Cytochromes content of mitochondria was measured by differential redox spectrophotometry, as described previously .

    Activation Assay:

    Article Title: Dynein light chain 1 induces assembly of large Bim complexes on mitochondria that stabilize Mcl-1 and regulate apoptosis
    Article Snippet: Paragraph title: Bax and Bak activation by cross-linking and flow cytometry ... For flow cytometry experiments, treated cells were fixed in 4% formalin for 15 min and stained with conformation-specific primary antibodies (Bax 6A7 [Sigma, no. B8429] and Bak NT [Millipore, no. 06536]).

    Staining:

    Article Title: Dynein light chain 1 induces assembly of large Bim complexes on mitochondria that stabilize Mcl-1 and regulate apoptosis
    Article Snippet: .. For flow cytometry experiments, treated cells were fixed in 4% formalin for 15 min and stained with conformation-specific primary antibodies (Bax 6A7 [Sigma, no. B8429] and Bak NT [Millipore, no. 06536]). .. Species-specific Alexa fluor 647 or Cy5-conjugated secondary antibodies were used to detect activated Bax and Bak by flow cytometry.

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  • 90
    Millipore mouse anti bax monoclonal
    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of <t>pro-caspase-8/9/3</t> (A) and <t>Bcl-2/Bax</t> (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P
    Mouse Anti Bax Monoclonal, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti bax monoclonal/product/Millipore
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    mouse anti bax monoclonal - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    90
    Millipore mouse monoclonal anti gapdh
    Effects of culture periods and culture temperatures on protein expressions for cytokeratins, <t>P63</t> and involucrin in ROE2 cells. The cells were cultured at different temperatures for 3, 6, and 9 days. Western blot analyses were performed with specific primary antibodies for Krt4, Krt10, Krt13, Krt14, Krt17/19, P63, involucrin, and <t>Gapdh.</t> Gapdh served as a loading control. Ctr, control (day 0).
    Mouse Monoclonal Anti Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 113 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti gapdh/product/Millipore
    Average 90 stars, based on 113 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal anti gapdh - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    90
    Millipore mouse anti gapdh
    Effects of culture periods and culture temperatures on protein expressions for cytokeratins, <t>P63</t> and involucrin in ROE2 cells. The cells were cultured at different temperatures for 3, 6, and 9 days. Western blot analyses were performed with specific primary antibodies for Krt4, Krt10, Krt13, Krt14, Krt17/19, P63, involucrin, and <t>Gapdh.</t> Gapdh served as a loading control. Ctr, control (day 0).
    Mouse Anti Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 428 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti gapdh/product/Millipore
    Average 90 stars, based on 428 article reviews
    Price from $9.99 to $1999.99
    mouse anti gapdh - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

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    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Cell Culture, Transfection, Western Blot, Positive Control

    Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Activation Assay, Expressing, Cell Culture, Transfection, Western Blot

    Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Western Blot

    Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of Gli-1 downregulation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two Gli-1-specific siRNAs (siGli-1-1 and siGli-1-2) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. Shh protein (5 μg/mL) was simultaneously added to the non-transfected LN229 cells as a positive control. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments ( n = 5). * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Cell Culture, Transfection, Western Blot, Positive Control

    Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Effects of mGluR4 activation on the expression of apoptosis-related proteins in LN229 cells. (A,B) Cultured LN229 cells were transfected with non-specific siRNA (siNC) and two mGluR4-specific siRNAs (simGluR4-1 and simGluR4-2) for 24 h, followed by treatment with the vehicle (Ctrl) or 30 μM of VU0155041 for 24 h. Then, the differential expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) , 9 (D) , 3 (E) , Bcl-2 (F) , Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) , each statistical value represents the mean ± SD of at least three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Activation Assay, Expressing, Cell Culture, Transfection, Western Blot

    Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Journal: Frontiers in Neuroscience

    Article Title: Activity of Metabotropic Glutamate Receptor 4 Suppresses Proliferation and Promotes Apoptosis With Inhibition of Gli-1 in Human Glioblastoma Cells

    doi: 10.3389/fnins.2018.00320

    Figure Lengend Snippet: Gli-1 downregulation is involved in mGluR4-mediated dynamic expression of apoptosis-related proteins in LN229 cells. (A,B) LN229 cells were treated with the vehicle (Ctrl), 5 μg/mL shh, or 30 μM VU0155041 plus 5 μg/mL shh (VU + shh) for 24 h. Then, the expression of pro-caspase-8/9/3 (A) and Bcl-2/Bax (B) was determined by western blot (WB) analysis. (C–H) WB bands were quantified to generate the ratios of pro-caspase-8 (C) ,−9 (D) , and−3 (E) , Bcl-2 (F) , and Bax (G) to β-actin and the ratio of Bcl-2 to Bax (H) ; each statistical value represents the mean ± SD of three independent experiments. * P

    Article Snippet: The primary antibodies and dilutions used in the experiments were as follows: rabbit anti-mGluR4 (1:1,000, Abcam); rabbit anti-Gli-1 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 3 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 8 polyclonal (1:1,000, Cell Signaling Technology); rabbit anti-caspase 9 polyclonal (1:1,000, Cell Signaling Technology); mouse anti-Bcl-2 monoclonal (1:1,000, Millipore); mouse anti-Bax monoclonal (1:1,000, Millipore); mouse anti-cyclin D1 monoclonal (1:1,000, Cell Signaling Technology); mouse anti-β-actin monoclonal (1:10,000, Sigma-Aldrich).

    Techniques: Expressing, Western Blot

    Effects of culture periods and culture temperatures on protein expressions for cytokeratins, P63 and involucrin in ROE2 cells. The cells were cultured at different temperatures for 3, 6, and 9 days. Western blot analyses were performed with specific primary antibodies for Krt4, Krt10, Krt13, Krt14, Krt17/19, P63, involucrin, and Gapdh. Gapdh served as a loading control. Ctr, control (day 0).

    Journal: Experimental Animals

    Article Title: Development of Oral Epithelial Cell Line ROE2 with Differentiation Potential from Transgenic Rats Harboring Temperature-Sensitive Simian Virus40 Large T-Antigen Gene

    doi: 10.1538/expanim.63.31

    Figure Lengend Snippet: Effects of culture periods and culture temperatures on protein expressions for cytokeratins, P63 and involucrin in ROE2 cells. The cells were cultured at different temperatures for 3, 6, and 9 days. Western blot analyses were performed with specific primary antibodies for Krt4, Krt10, Krt13, Krt14, Krt17/19, P63, involucrin, and Gapdh. Gapdh served as a loading control. Ctr, control (day 0).

    Article Snippet: Primary antibodies used were as follows: rabbit polyclonal anti-Krt4 antibody (ab51600, Abcam); mouse monoclonal anti-Krt10 antibody (ab9025, Abcam); mouse monoclonal anti-Krt13 antibody (1C7, MUbio Products BV, Oxfordlaan, Maastricht, The Netherlands); mouse monoclonal anti-Krt14 antibody (Millipore Co.); rabbit monoclonal anti-Krt17/19 antibody (#3984, Cell Signaling Technology, Inc.); mouse monoclonal anti-P63 antibody (sc-8431, Santa Cruz Biotechnology); mouse monoclonal anti-involucrin antibody (ab80530, Abcam); mouse monoclonal anti-Gapdh (glyceraldehyde 3-phosphate dehydrogenase) antibody (MAB374, Millipore Co.).

    Techniques: Cell Culture, Western Blot