Structured Review

Merck KGaA glyceraldehyde 3 phosphate dehydrogenase
Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related <t>glyceraldehyde-3-phosphate</t> dehydrogenase. The results are represented as means ± SEM ( n = 4). # P
Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/glyceraldehyde 3 phosphate dehydrogenase/product/Merck KGaA
Average 94 stars, based on 1 article reviews
Price from $9.99 to $1999.99
glyceraldehyde 3 phosphate dehydrogenase - by Bioz Stars, 2020-04
94/100 stars

Images

1) Product Images from "Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway"

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2019/1354345

Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related glyceraldehyde-3-phosphate dehydrogenase. The results are represented as means ± SEM ( n = 4). # P
Figure Legend Snippet: Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related glyceraldehyde-3-phosphate dehydrogenase. The results are represented as means ± SEM ( n = 4). # P

Techniques Used: Expressing, Mouse Assay

Related Articles

Synthesized:

Article Title: AMPK antagonizes hepatic glucagon-stimulated cyclic AMP signalling via phosphorylation-induced activation of cyclic nucleotide phosphodiesterase 4B
Article Snippet: Anti-total ACC (Merck Millipore, Catalogue No. 04-322), anti-P-Ser79-ACC (Merck-Millipore, Catalogue No. 07-303), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Catalogue No. MAB374), anti-total GP (Sigma, Catalogue No. HPA000962), anti-total AMPK β1(R & D Systems, Catalogue No. AF2854) and anti-total AMPK β2 (R & D Systems, Catalogue No. MAB3808), anti-PThr172-AMPK (T172) (Cell Signaling Technologies, Catalogue No. 2535, anti-P-AMPK-substrate (Cell Signaling Technologies, Catalogue No. 5759), anti-P-PKA-substrate (Cell Signaling Technologies, Catalogue No. 9624), anti-total Raptor (Cell Signaling Technologies, Catalogue No. 2280) and anti-P-Ser792-Raptor (Cell Signaling Technologies, Catalogue No. 2083), anti-total CREB (Cell Signaling Technologies, Catalogue No. 9197), anti-phospho-Ser133-CREB (Cell Signaling Technologies, Catalogue No. 9198) and anti-total PDE4B (Origene, Catalogue No. TA503471) antibodies were from the sources cited. .. A peptide surrounding Ser304 of mouse PDE4B (CKLMHSS S LNNTSI) was synthesized with or without Ser304 phosphorylated and with a Cys (N-term) for coupling to keyhole limpet haemocyanin (KLH) or bovine serum albumin (BSA) (Imject maleimide-activated KLH/BSA kit, Thermo Fisher Scientific).

Affinity Purification:

Article Title: AMPK antagonizes hepatic glucagon-stimulated cyclic AMP signalling via phosphorylation-induced activation of cyclic nucleotide phosphodiesterase 4B
Article Snippet: Anti-total ACC (Merck Millipore, Catalogue No. 04-322), anti-P-Ser79-ACC (Merck-Millipore, Catalogue No. 07-303), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Catalogue No. MAB374), anti-total GP (Sigma, Catalogue No. HPA000962), anti-total AMPK β1(R & D Systems, Catalogue No. AF2854) and anti-total AMPK β2 (R & D Systems, Catalogue No. MAB3808), anti-PThr172-AMPK (T172) (Cell Signaling Technologies, Catalogue No. 2535, anti-P-AMPK-substrate (Cell Signaling Technologies, Catalogue No. 5759), anti-P-PKA-substrate (Cell Signaling Technologies, Catalogue No. 9624), anti-total Raptor (Cell Signaling Technologies, Catalogue No. 2280) and anti-P-Ser792-Raptor (Cell Signaling Technologies, Catalogue No. 2083), anti-total CREB (Cell Signaling Technologies, Catalogue No. 9197), anti-phospho-Ser133-CREB (Cell Signaling Technologies, Catalogue No. 9198) and anti-total PDE4B (Origene, Catalogue No. TA503471) antibodies were from the sources cited. .. The KLH-coupled phosphopeptide was injected in rabbits (Thermo Fisher Scientific), and the serum was affinity purified on both BSA-coupled phosphopeptide and non-phosphopeptide linked to CH-activated Sepharose 4B (GE Healthcare).

Blocking Assay:

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Membranes were blocked for 1 hour in Odyssey blocking buffer (Li-Cor Biosciences UK Ltd., Cambridge, UK), incubated with titrated primary antibody overnight at 4°C. .. Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution).

Article Title: The Antidiabetic and Antinephritic Activities of Tuber melanosporum via Modulation of Nrf2-Mediated Oxidative Stress in the db/db Mouse
Article Snippet: .. After blocking with 5% BSA for 4 h, the transferred membranes were incubated with the following primary antibodies overnight at 4°C at a dilution of 1 : 2000: phosphor-janus kinase 2 (p-JAK2, ab68268), total-janus kinase 2 (t-JAK2, ab39636), phosphor-nuclear factor-κ B (p-NF-κ B, ab86299), total-NF-κ B (t-NF-κ B, ab32536), phosphor-signal transducer and activator of transcription 3 (p-STAT3, ab76315), total-STAT3 (t-STAT3, M06-596), catalase (CAT, ab16731), heme oxygenase 1 (HO-1, ab68477), nuclear respiratory factor 2 (Nrf2, ab137550), manganese superoxide dismutase 2 (SOD2, ab13533), heme oxygenase 2 (HO-2, ab90492), protein kinase C alpha (PKC-α , ab23513) (Abcam, Cambridge, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16) (Merck Millipore, Darmstadt, Germany). .. After 5 washes with TBST buffer, the transferred membranes were incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (sc-3836) (Santa Cruz Biotechnology, Santa Cruz, USA) for 4 h at 4°C.

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: .. After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C. .. The membrane was then washed with PBS containing 0.05% Tween-20 and incubated with anti-mouse (IH-0031) and anti-rabbit (IH-0011) (Dingguo Biotechnology Co., Ltd., Beijing, China) horseradish peroxidase-conjugated secondary antibodies for 4 h at 4°C.

Electrophoresis:

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Protein samples (20 to 40 μg) were resolved by electrophoresis of NuPAGE gels using the Novex system (Invitrogen) and electrotransferred to polyvinylidene difluoride membranes (Merck Millipore). .. Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution).

Incubation:

Article Title: BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo
Article Snippet: .. To confirm the presence of equal amounts of loaded proteins, membranes were incubated with anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, clone 6C5) (Merck Millipore, Guyancourt, France). .. Signals were visualized by chemiluminescence and processed by the Image LabTM software (BioRad, Marnes-la Coquette, France).

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: .. Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C. .. Specific signals were visualized with ECL detection on a gel imaging system (UVP, California, USA).

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Membranes were blocked for 1 hour in Odyssey blocking buffer (Li-Cor Biosciences UK Ltd., Cambridge, UK), incubated with titrated primary antibody overnight at 4°C. .. Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution).

Article Title: The Antidiabetic and Antinephritic Activities of Tuber melanosporum via Modulation of Nrf2-Mediated Oxidative Stress in the db/db Mouse
Article Snippet: .. After blocking with 5% BSA for 4 h, the transferred membranes were incubated with the following primary antibodies overnight at 4°C at a dilution of 1 : 2000: phosphor-janus kinase 2 (p-JAK2, ab68268), total-janus kinase 2 (t-JAK2, ab39636), phosphor-nuclear factor-κ B (p-NF-κ B, ab86299), total-NF-κ B (t-NF-κ B, ab32536), phosphor-signal transducer and activator of transcription 3 (p-STAT3, ab76315), total-STAT3 (t-STAT3, M06-596), catalase (CAT, ab16731), heme oxygenase 1 (HO-1, ab68477), nuclear respiratory factor 2 (Nrf2, ab137550), manganese superoxide dismutase 2 (SOD2, ab13533), heme oxygenase 2 (HO-2, ab90492), protein kinase C alpha (PKC-α , ab23513) (Abcam, Cambridge, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16) (Merck Millipore, Darmstadt, Germany). .. After 5 washes with TBST buffer, the transferred membranes were incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (sc-3836) (Santa Cruz Biotechnology, Santa Cruz, USA) for 4 h at 4°C.

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: .. After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C. .. The membrane was then washed with PBS containing 0.05% Tween-20 and incubated with anti-mouse (IH-0031) and anti-rabbit (IH-0011) (Dingguo Biotechnology Co., Ltd., Beijing, China) horseradish peroxidase-conjugated secondary antibodies for 4 h at 4°C.

Article Title: Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c
Article Snippet: .. For immunoblot detection, a nitrocellulose membrane was incubated with the following primary antibodies: anti-smad family member 2 (smad2; Cell Signaling Technology Inc., MA, USA), anti-phospho smad2 (p-smad2; Cell Signaling Technology Inc.) anti-zinc finger E-box binding homeobox 1 (zeb1; Santa Cruz Biotechnology, TX,USA), anti-epithelial cadherin (E-cadherin; Becton Dickinson Co., NJ, USA), anti-vimentin (Cell Signaling Technology Inc.), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck Millipore Co., MA, USA) and anti-fibrillarin (Abcam, Cambridge, UK). .. The membrane was then incubated with secondary antibody and detected by enhanced chemiluminescence (Pierce, IL, USA) with visualization by exposure to Amersham Hyperfilm (Amersham Life Sciences, IL, USA).

Expressing:

Article Title: Modulation of renal calcium and phosphate transporting proteins by dietary nitrogen and/or calcium in young goats
Article Snippet: .. For CaBPD28K , glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) was used as internal standard with constant expression level (anti-GAPDH; Merck Millipore), and for Na+ /K+ -ATPase and PTHR, signal intensities per millimeter were measured for determining relative amounts of protein expression. ..

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: For detecting the expression levels of Nrf2 in the nucleus of liver tissue, the liver tissues were homogenized using the Cytoplasmic Extraction Reagent I (Thermo, Rockford, USA) following with the incubation of the Cytoplasmic Extraction Reagent II (Thermo, Rockford, USA) for 10 min to obtain the cytoplasmic protein extraction, and then the collected precipitate was incubated with a Nuclear Extraction Reagent (Thermo, Rockford, USA) for another 40 minutes to obtain nuclear protein. .. After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C.

BIA-KA:

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: The protein concentrations of the homogenates were measured using the BCA Protein Assay Kit (Merck Millipore, USA); 50 μ g of protein was electrophoresed on 12% SDS-PAGE, transferred onto polyvinylidene difluoride (PVDF) membrane (Merck Millipore, USA), and blocked in 5% bovine serum albumin (BSA) in Tris-buffered saline. .. Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C.

Western Blot:

Article Title: BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo
Article Snippet: Paragraph title: Western blotting ... To confirm the presence of equal amounts of loaded proteins, membranes were incubated with anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, clone 6C5) (Merck Millipore, Guyancourt, France).

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: Paragraph title: 2.8. Western Blot Analysis ... Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C.

Article Title: Calpain cleavage within dysferlin exon 40a releases a synaptotagmin-like module for membrane repair
Article Snippet: Paragraph title: Western blot. ... We used Hamlet 1 (1:500; Leica Microsystems, Wetzlar, Germany; ), Romeo (1:1000; Abcam; ), mitsugumin-53/TRIM72 (1:5000; a generous gift from Jianjie Ma, University of Medicine and Dentistry of New Jersey, Piscataway, NJ; ), c-Myc (1:500; Santa Cruz Biotechnology, Dallas, TX), β-tubulin (1:1000; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH: 1:5000; Merck, Millipore), OTOF-C12 (Santa Cruz Biotechnology), and anti-myoferlin 7D2 (Abcam; ).

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Paragraph title: Western Blot Analysis ... Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution).

Article Title: The Antidiabetic and Antinephritic Activities of Tuber melanosporum via Modulation of Nrf2-Mediated Oxidative Stress in the db/db Mouse
Article Snippet: Paragraph title: 2.6. Western Blot ... After blocking with 5% BSA for 4 h, the transferred membranes were incubated with the following primary antibodies overnight at 4°C at a dilution of 1 : 2000: phosphor-janus kinase 2 (p-JAK2, ab68268), total-janus kinase 2 (t-JAK2, ab39636), phosphor-nuclear factor-κ B (p-NF-κ B, ab86299), total-NF-κ B (t-NF-κ B, ab32536), phosphor-signal transducer and activator of transcription 3 (p-STAT3, ab76315), total-STAT3 (t-STAT3, M06-596), catalase (CAT, ab16731), heme oxygenase 1 (HO-1, ab68477), nuclear respiratory factor 2 (Nrf2, ab137550), manganese superoxide dismutase 2 (SOD2, ab13533), heme oxygenase 2 (HO-2, ab90492), protein kinase C alpha (PKC-α , ab23513) (Abcam, Cambridge, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16) (Merck Millipore, Darmstadt, Germany).

Article Title: Hyperosmotic stress activates the expression of members of the miR-15/107 family and induces downregulation of anti-apoptotic genes in rat liver
Article Snippet: Material The Bcl2 recognizing antibody (#ab7973) was purchased from abcam (Cambridge, UK) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; #MAB374) from Merck-Millipore (Darmstadt, Germany). .. Antibodies recognizing CD95 (#sc-715; IP) were from Santa Cruz Biotechnology (Heidelberg, Germany), phospho-tyrosine (#05-321) from Merck-Millipore (Darmstadt, Germany), CD95 (#MA1-7622; WB) from Life Technologies GmbH (Darmstadt, Germany), cleaved caspase 3 (#9664), γ-tubulin (#T5326) from Sigma Aldrich (Munich, Germany).

Article Title: Cancer-associated fibroblast-derived Gremlin 1 promotes breast cancer progression
Article Snippet: Paragraph title: Western blotting ... The antibodies used for immunoblotting were raised against the following proteins: phospho-SMAD1/5/8 (pSMAD1/5/8, home-made) [ ], αSMA (A2547, Sigma-Aldrich), fibronectin (F7387, Sigma-Aldrich), FAP (WH0002191M1, Sigma-Aldrich), collagen I (ab34710, Abcam), vimentin (5741, Cell signaling), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, MAB374, Merck Millipore).

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: Paragraph title: 2.6. Western Blotting ... After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C.

Article Title: Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c
Article Snippet: Paragraph title: Sample preparation and western blotting ... For immunoblot detection, a nitrocellulose membrane was incubated with the following primary antibodies: anti-smad family member 2 (smad2; Cell Signaling Technology Inc., MA, USA), anti-phospho smad2 (p-smad2; Cell Signaling Technology Inc.) anti-zinc finger E-box binding homeobox 1 (zeb1; Santa Cruz Biotechnology, TX,USA), anti-epithelial cadherin (E-cadherin; Becton Dickinson Co., NJ, USA), anti-vimentin (Cell Signaling Technology Inc.), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck Millipore Co., MA, USA) and anti-fibrillarin (Abcam, Cambridge, UK).

Colorimetric Assay:

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Lysates were sonicated and microcentrifuged, and the protein concentration was determined by Bradford colorimetric assay. .. Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution).

Protease Inhibitor:

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: Western Blot Analysis Partial kidney tissues were thoroughly homogenized in a lysis buffer (0.97% protease inhibitor cocktail, 0.94% 50 mM phenylmethylsulfonyl fluoride, and 97.09% 1x RIPA) on ice. .. Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C.

Article Title: The Antidiabetic and Antinephritic Activities of Tuber melanosporum via Modulation of Nrf2-Mediated Oxidative Stress in the db/db Mouse
Article Snippet: Western Blot One part of the collected kidney tissue was homogenized in RIPA containing 1% protease inhibitor cocktail and 2% phenylmethanesulfonyl fluoride. .. After blocking with 5% BSA for 4 h, the transferred membranes were incubated with the following primary antibodies overnight at 4°C at a dilution of 1 : 2000: phosphor-janus kinase 2 (p-JAK2, ab68268), total-janus kinase 2 (t-JAK2, ab39636), phosphor-nuclear factor-κ B (p-NF-κ B, ab86299), total-NF-κ B (t-NF-κ B, ab32536), phosphor-signal transducer and activator of transcription 3 (p-STAT3, ab76315), total-STAT3 (t-STAT3, M06-596), catalase (CAT, ab16731), heme oxygenase 1 (HO-1, ab68477), nuclear respiratory factor 2 (Nrf2, ab137550), manganese superoxide dismutase 2 (SOD2, ab13533), heme oxygenase 2 (HO-2, ab90492), protein kinase C alpha (PKC-α , ab23513) (Abcam, Cambridge, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16) (Merck Millipore, Darmstadt, Germany).

Article Title: Cancer-associated fibroblast-derived Gremlin 1 promotes breast cancer progression
Article Snippet: Western blotting Cells were lysed with RIPA buffer containing 1 × complete protease inhibitor cocktail (11836153001, Roche). .. The antibodies used for immunoblotting were raised against the following proteins: phospho-SMAD1/5/8 (pSMAD1/5/8, home-made) [ ], αSMA (A2547, Sigma-Aldrich), fibronectin (F7387, Sigma-Aldrich), FAP (WH0002191M1, Sigma-Aldrich), collagen I (ab34710, Abcam), vimentin (5741, Cell signaling), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, MAB374, Merck Millipore).

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: Western Blotting A portion of the liver from each mouse was homogenized in cell lysis buffer containing 1% protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA), 2% phenylmethanesulfonyl fluoride (Sigma-Aldrich, St. Louis, MO, USA), and 97% RIPA (Sigma-Aldrich, St. Louis, MO, USA). .. After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C.

Article Title: Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c
Article Snippet: Sample preparation and western blotting HCC827 or HCC827GR cells were solubilized with buffer containing 20 mM Tris-HCl (pH7.4), 0.3%(w/v) Triton, 3 mM MgCl2 , 1 M sucrose, 5 mM α-ME, and 1/1,000 protease inhibitor or M-PER Mammalian Protein Extraction Reagent (Thermo Fisher Scientific Inc.), and cell lysates were prepared as described previously [ ]. .. For immunoblot detection, a nitrocellulose membrane was incubated with the following primary antibodies: anti-smad family member 2 (smad2; Cell Signaling Technology Inc., MA, USA), anti-phospho smad2 (p-smad2; Cell Signaling Technology Inc.) anti-zinc finger E-box binding homeobox 1 (zeb1; Santa Cruz Biotechnology, TX,USA), anti-epithelial cadherin (E-cadherin; Becton Dickinson Co., NJ, USA), anti-vimentin (Cell Signaling Technology Inc.), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck Millipore Co., MA, USA) and anti-fibrillarin (Abcam, Cambridge, UK).

Imaging:

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C. .. Specific signals were visualized with ECL detection on a gel imaging system (UVP, California, USA).

Article Title: The Antidiabetic and Antinephritic Activities of Tuber melanosporum via Modulation of Nrf2-Mediated Oxidative Stress in the db/db Mouse
Article Snippet: After blocking with 5% BSA for 4 h, the transferred membranes were incubated with the following primary antibodies overnight at 4°C at a dilution of 1 : 2000: phosphor-janus kinase 2 (p-JAK2, ab68268), total-janus kinase 2 (t-JAK2, ab39636), phosphor-nuclear factor-κ B (p-NF-κ B, ab86299), total-NF-κ B (t-NF-κ B, ab32536), phosphor-signal transducer and activator of transcription 3 (p-STAT3, ab76315), total-STAT3 (t-STAT3, M06-596), catalase (CAT, ab16731), heme oxygenase 1 (HO-1, ab68477), nuclear respiratory factor 2 (Nrf2, ab137550), manganese superoxide dismutase 2 (SOD2, ab13533), heme oxygenase 2 (HO-2, ab90492), protein kinase C alpha (PKC-α , ab23513) (Abcam, Cambridge, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16) (Merck Millipore, Darmstadt, Germany). .. The intensity of the bands was quantified by scanning densitometry using an imaging system (Ultra-Violet Products Ltd., Cambridge, UK).

Article Title: Cancer-associated fibroblast-derived Gremlin 1 promotes breast cancer progression
Article Snippet: The signal was detected using Clarity™ Western ECL Substrate (1705060, Bio-Rad) and ChemiDoc Imaging System (17001402, Bio-Rad). .. The antibodies used for immunoblotting were raised against the following proteins: phospho-SMAD1/5/8 (pSMAD1/5/8, home-made) [ ], αSMA (A2547, Sigma-Aldrich), fibronectin (F7387, Sigma-Aldrich), FAP (WH0002191M1, Sigma-Aldrich), collagen I (ab34710, Abcam), vimentin (5741, Cell signaling), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, MAB374, Merck Millipore).

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C. .. After three washes, the target proteins were visualized using an Immobilon Western Chemiluminescent HRP substrate (WBKLS0500; Merck Millipore, Burlington, Massachusetts, USA) and a gel imaging system (Biodoc IT2 315, Analytik Jena, Jena, Germany).

Protein Concentration:

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Lysates were sonicated and microcentrifuged, and the protein concentration was determined by Bradford colorimetric assay. .. Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution).

Sonication:

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Lysates were sonicated and microcentrifuged, and the protein concentration was determined by Bradford colorimetric assay. .. Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution).

Injection:

Article Title: AMPK antagonizes hepatic glucagon-stimulated cyclic AMP signalling via phosphorylation-induced activation of cyclic nucleotide phosphodiesterase 4B
Article Snippet: Anti-total ACC (Merck Millipore, Catalogue No. 04-322), anti-P-Ser79-ACC (Merck-Millipore, Catalogue No. 07-303), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck-Millipore, Catalogue No. MAB374), anti-total GP (Sigma, Catalogue No. HPA000962), anti-total AMPK β1(R & D Systems, Catalogue No. AF2854) and anti-total AMPK β2 (R & D Systems, Catalogue No. MAB3808), anti-PThr172-AMPK (T172) (Cell Signaling Technologies, Catalogue No. 2535, anti-P-AMPK-substrate (Cell Signaling Technologies, Catalogue No. 5759), anti-P-PKA-substrate (Cell Signaling Technologies, Catalogue No. 9624), anti-total Raptor (Cell Signaling Technologies, Catalogue No. 2280) and anti-P-Ser792-Raptor (Cell Signaling Technologies, Catalogue No. 2083), anti-total CREB (Cell Signaling Technologies, Catalogue No. 9197), anti-phospho-Ser133-CREB (Cell Signaling Technologies, Catalogue No. 9198) and anti-total PDE4B (Origene, Catalogue No. TA503471) antibodies were from the sources cited. .. The KLH-coupled phosphopeptide was injected in rabbits (Thermo Fisher Scientific), and the serum was affinity purified on both BSA-coupled phosphopeptide and non-phosphopeptide linked to CH-activated Sepharose 4B (GE Healthcare).

Binding Assay:

Article Title: Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c
Article Snippet: .. For immunoblot detection, a nitrocellulose membrane was incubated with the following primary antibodies: anti-smad family member 2 (smad2; Cell Signaling Technology Inc., MA, USA), anti-phospho smad2 (p-smad2; Cell Signaling Technology Inc.) anti-zinc finger E-box binding homeobox 1 (zeb1; Santa Cruz Biotechnology, TX,USA), anti-epithelial cadherin (E-cadherin; Becton Dickinson Co., NJ, USA), anti-vimentin (Cell Signaling Technology Inc.), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck Millipore Co., MA, USA) and anti-fibrillarin (Abcam, Cambridge, UK). .. The membrane was then incubated with secondary antibody and detected by enhanced chemiluminescence (Pierce, IL, USA) with visualization by exposure to Amersham Hyperfilm (Amersham Life Sciences, IL, USA).

Nucleic Acid Electrophoresis:

Article Title: BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo
Article Snippet: Lysates were centrifuged for 10 minutes at 300 g at 4 °C, and supernatants subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred electrophoretically to polyvinylidene difluoride (PVDF) membranes. .. To confirm the presence of equal amounts of loaded proteins, membranes were incubated with anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, clone 6C5) (Merck Millipore, Guyancourt, France).

Bicinchoninic Acid Protein Assay:

Article Title: Cancer-associated fibroblast-derived Gremlin 1 promotes breast cancer progression
Article Snippet: Protein concentrations were determined using a bicinchoninic acid protein assay kit (5000111, Bio-Rad). .. The antibodies used for immunoblotting were raised against the following proteins: phospho-SMAD1/5/8 (pSMAD1/5/8, home-made) [ ], αSMA (A2547, Sigma-Aldrich), fibronectin (F7387, Sigma-Aldrich), FAP (WH0002191M1, Sigma-Aldrich), collagen I (ab34710, Abcam), vimentin (5741, Cell signaling), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, MAB374, Merck Millipore).

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: The total protein concentrations were measured using a bicinchoninic acid (BCA) protein assay kit (BCA; Merck Millipore, Burlington, Massachusetts, USA). .. After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C.

Labeling:

Article Title: Ketamine-Induced Apoptosis in Normal Human Urothelial Cells
Article Snippet: Densitometry was normalized using the intensity of housekeeping proteins β-actin (Sigma-Aldrich; clone AC15, mouse; 1:10,000 dilution) or glyceraldehyde-3-phosphate dehydrogenase (Merck Millipore; clone 6C5, mouse; 1:2000 dilution). .. Membranes were labeled with the appropriate IRDye-conjugated secondary antibody and visualized using an Odyssey Infrared imager (Li-Cor).

Protein Extraction:

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: For detecting the expression levels of Nrf2 in the nucleus of liver tissue, the liver tissues were homogenized using the Cytoplasmic Extraction Reagent I (Thermo, Rockford, USA) following with the incubation of the Cytoplasmic Extraction Reagent II (Thermo, Rockford, USA) for 10 min to obtain the cytoplasmic protein extraction, and then the collected precipitate was incubated with a Nuclear Extraction Reagent (Thermo, Rockford, USA) for another 40 minutes to obtain nuclear protein. .. After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C.

Article Title: Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c
Article Snippet: Sample preparation and western blotting HCC827 or HCC827GR cells were solubilized with buffer containing 20 mM Tris-HCl (pH7.4), 0.3%(w/v) Triton, 3 mM MgCl2 , 1 M sucrose, 5 mM α-ME, and 1/1,000 protease inhibitor or M-PER Mammalian Protein Extraction Reagent (Thermo Fisher Scientific Inc.), and cell lysates were prepared as described previously [ ]. .. For immunoblot detection, a nitrocellulose membrane was incubated with the following primary antibodies: anti-smad family member 2 (smad2; Cell Signaling Technology Inc., MA, USA), anti-phospho smad2 (p-smad2; Cell Signaling Technology Inc.) anti-zinc finger E-box binding homeobox 1 (zeb1; Santa Cruz Biotechnology, TX,USA), anti-epithelial cadherin (E-cadherin; Becton Dickinson Co., NJ, USA), anti-vimentin (Cell Signaling Technology Inc.), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck Millipore Co., MA, USA) and anti-fibrillarin (Abcam, Cambridge, UK).

Polyacrylamide Gel Electrophoresis:

Article Title: Cancer-associated fibroblast-derived Gremlin 1 promotes breast cancer progression
Article Snippet: Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto 45-μm polyvinylidene difluoride (PVDF) membrane (IPVH00010, Merck Millipore). .. The antibodies used for immunoblotting were raised against the following proteins: phospho-SMAD1/5/8 (pSMAD1/5/8, home-made) [ ], αSMA (A2547, Sigma-Aldrich), fibronectin (F7387, Sigma-Aldrich), FAP (WH0002191M1, Sigma-Aldrich), collagen I (ab34710, Abcam), vimentin (5741, Cell signaling), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, MAB374, Merck Millipore).

IA:

Article Title: Calpain cleavage within dysferlin exon 40a releases a synaptotagmin-like module for membrane repair
Article Snippet: .. We used Hamlet 1 (1:500; Leica Microsystems, Wetzlar, Germany; ), Romeo (1:1000; Abcam; ), mitsugumin-53/TRIM72 (1:5000; a generous gift from Jianjie Ma, University of Medicine and Dentistry of New Jersey, Piscataway, NJ; ), c-Myc (1:500; Santa Cruz Biotechnology, Dallas, TX), β-tubulin (1:1000; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH: 1:5000; Merck, Millipore), OTOF-C12 (Santa Cruz Biotechnology), and anti-myoferlin 7D2 (Abcam; ). ..

Chloramphenicol Acetyltransferase Assay:

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: .. Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C. .. Specific signals were visualized with ECL detection on a gel imaging system (UVP, California, USA).

Article Title: The Antidiabetic and Antinephritic Activities of Tuber melanosporum via Modulation of Nrf2-Mediated Oxidative Stress in the db/db Mouse
Article Snippet: .. After blocking with 5% BSA for 4 h, the transferred membranes were incubated with the following primary antibodies overnight at 4°C at a dilution of 1 : 2000: phosphor-janus kinase 2 (p-JAK2, ab68268), total-janus kinase 2 (t-JAK2, ab39636), phosphor-nuclear factor-κ B (p-NF-κ B, ab86299), total-NF-κ B (t-NF-κ B, ab32536), phosphor-signal transducer and activator of transcription 3 (p-STAT3, ab76315), total-STAT3 (t-STAT3, M06-596), catalase (CAT, ab16731), heme oxygenase 1 (HO-1, ab68477), nuclear respiratory factor 2 (Nrf2, ab137550), manganese superoxide dismutase 2 (SOD2, ab13533), heme oxygenase 2 (HO-2, ab90492), protein kinase C alpha (PKC-α , ab23513) (Abcam, Cambridge, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16) (Merck Millipore, Darmstadt, Germany). .. After 5 washes with TBST buffer, the transferred membranes were incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (sc-3836) (Santa Cruz Biotechnology, Santa Cruz, USA) for 4 h at 4°C.

SDS Page:

Article Title: BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo
Article Snippet: Lysates were centrifuged for 10 minutes at 300 g at 4 °C, and supernatants subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred electrophoretically to polyvinylidene difluoride (PVDF) membranes. .. To confirm the presence of equal amounts of loaded proteins, membranes were incubated with anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, clone 6C5) (Merck Millipore, Guyancourt, France).

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: The protein concentrations of the homogenates were measured using the BCA Protein Assay Kit (Merck Millipore, USA); 50 μ g of protein was electrophoresed on 12% SDS-PAGE, transferred onto polyvinylidene difluoride (PVDF) membrane (Merck Millipore, USA), and blocked in 5% bovine serum albumin (BSA) in Tris-buffered saline. .. Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C.

Article Title: The Antidiabetic and Antinephritic Activities of Tuber melanosporum via Modulation of Nrf2-Mediated Oxidative Stress in the db/db Mouse
Article Snippet: Protein concentrations were determined by the Bradford method, and 40 μ g of protein was separated using 12% SDS-PAGE gel and electroblotted onto a nitrocellulose membrane (0.45 μ m; Bio Basic Inc., USA). .. After blocking with 5% BSA for 4 h, the transferred membranes were incubated with the following primary antibodies overnight at 4°C at a dilution of 1 : 2000: phosphor-janus kinase 2 (p-JAK2, ab68268), total-janus kinase 2 (t-JAK2, ab39636), phosphor-nuclear factor-κ B (p-NF-κ B, ab86299), total-NF-κ B (t-NF-κ B, ab32536), phosphor-signal transducer and activator of transcription 3 (p-STAT3, ab76315), total-STAT3 (t-STAT3, M06-596), catalase (CAT, ab16731), heme oxygenase 1 (HO-1, ab68477), nuclear respiratory factor 2 (Nrf2, ab137550), manganese superoxide dismutase 2 (SOD2, ab13533), heme oxygenase 2 (HO-2, ab90492), protein kinase C alpha (PKC-α , ab23513) (Abcam, Cambridge, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16) (Merck Millipore, Darmstadt, Germany).

Article Title: Cancer-associated fibroblast-derived Gremlin 1 promotes breast cancer progression
Article Snippet: Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto 45-μm polyvinylidene difluoride (PVDF) membrane (IPVH00010, Merck Millipore). .. The antibodies used for immunoblotting were raised against the following proteins: phospho-SMAD1/5/8 (pSMAD1/5/8, home-made) [ ], αSMA (A2547, Sigma-Aldrich), fibronectin (F7387, Sigma-Aldrich), FAP (WH0002191M1, Sigma-Aldrich), collagen I (ab34710, Abcam), vimentin (5741, Cell signaling), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, MAB374, Merck Millipore).

Software:

Article Title: BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo
Article Snippet: To confirm the presence of equal amounts of loaded proteins, membranes were incubated with anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, clone 6C5) (Merck Millipore, Guyancourt, France). .. Signals were visualized by chemiluminescence and processed by the Image LabTM software (BioRad, Marnes-la Coquette, France).

Article Title: Modulation of renal calcium and phosphate transporting proteins by dietary nitrogen and/or calcium in young goats
Article Snippet: Densitometric measurements of proteins were performed using Image Lab 5.2.1 software (Bio-Rad). .. For CaBPD28K , glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) was used as internal standard with constant expression level (anti-GAPDH; Merck Millipore), and for Na+ /K+ -ATPase and PTHR, signal intensities per millimeter were measured for determining relative amounts of protein expression.

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C. .. The bands were quantitated using Image J analysis software, version 1.46 (National Institutes of Health, Bethesda, MD, USA).

Sample Prep:

Article Title: Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c
Article Snippet: Paragraph title: Sample preparation and western blotting ... For immunoblot detection, a nitrocellulose membrane was incubated with the following primary antibodies: anti-smad family member 2 (smad2; Cell Signaling Technology Inc., MA, USA), anti-phospho smad2 (p-smad2; Cell Signaling Technology Inc.) anti-zinc finger E-box binding homeobox 1 (zeb1; Santa Cruz Biotechnology, TX,USA), anti-epithelial cadherin (E-cadherin; Becton Dickinson Co., NJ, USA), anti-vimentin (Cell Signaling Technology Inc.), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Merck Millipore Co., MA, USA) and anti-fibrillarin (Abcam, Cambridge, UK).

Lysis:

Article Title: BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo
Article Snippet: Western blotting After stimulation, B cells were centrifuged and cell pellets re-suspended in lysis buffer (1% Triton X-100, 20 mM Tris-HCl [pH 8], 130 mM NaCl, 10% glycerol, 2 mM EDTA, 1 mM PMSF, and protease inhibitors) for 20 minutes on ice. .. To confirm the presence of equal amounts of loaded proteins, membranes were incubated with anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, clone 6C5) (Merck Millipore, Guyancourt, France).

Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway
Article Snippet: Western Blot Analysis Partial kidney tissues were thoroughly homogenized in a lysis buffer (0.97% protease inhibitor cocktail, 0.94% 50 mM phenylmethylsulfonyl fluoride, and 97.09% 1x RIPA) on ice. .. Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C.

Article Title: Aronia melanocarpa Prevents Alcohol-Induced Chronic Liver Injury via Regulation of Nrf2 Signaling in C57BL/6 Mice
Article Snippet: Western Blotting A portion of the liver from each mouse was homogenized in cell lysis buffer containing 1% protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA), 2% phenylmethanesulfonyl fluoride (Sigma-Aldrich, St. Louis, MO, USA), and 97% RIPA (Sigma-Aldrich, St. Louis, MO, USA). .. After blocking with 5% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 4 h, the membrane was incubated with primary antibody (1 : 1000 dilution) against glyceraldehyde-3-phosphate dehydrogenase (GAPDH; ABS16), total-signal transducer and activator of transcription 3 (T-Stat3; 06-596) (Merck Millipore, Burlington, Massachusetts, USA), beta-actin (β -actin; sc-47778) (Santa Cruz, 2145 Delaware Ave, USA), phosphor- (P-) Stat3 (cstD3A7), P-phosphatidylinositol-3-hydroxykinase (PI3K; cst4228s), T-PI3K (cst4292s; Cell Signaling Technology, Boston, MA, USA), P-protein kinase B (P-Akt) (ab108266), T-Akt (ab200195), Kelch-like ECH-associated protein 1 (Keap1; ab150654), Nrf2 (ab89443), heme oxygenase-1 (HO-1; ab137749), SOD-2 (ab13533) (Abcam, Cambridge Science Park, UK), HO-2 (bs-1238R), SOD-1 (bs-10216R), and lamin B (bs-1840R) (Bioss Antibodies Biotechnology Co., Ltd., Beijing, China) overnight at 4°C.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 80
    Merck KGaA anti bax
    Inhibition of endothelial cell conditioned medium (EC-CM) induced pro-apoptotic molecule expression by isoflurane. To examine the effect of pretreated isoflurane on neuronal cell survival and death under culture with EC-CM, we performed western blot analysis using <t>Bcl-2</t> and <t>Bax.</t> (B) The Bcl-2 protein was significantly upregulated in neuronal cells incubated with EC-CM of isoflurane, RAP, or isoflurane RAP treated groups in the tPA and OGD/R condition. (C) In contrast to Bcl-2, the protein levels of Bax was significantly downregulated in neuronal cells incubated with EC-CM of isoflurane, RAP or isoflurane + RAP-treated groups in the tPA and OGD/R condition. (E) Bcl-2 protein levels were significantly increased in neuronal cells incubated with EC-CM of isoflurane, MG-132, or isoflurane + MG-132-treated groups in the tPA and OGD/R condition. (F) Bax was significantly decreased in neuronal cells incubated with EC-CM of isoflurane, MG-132 and isoflurane + MG-132-treated groups in the tPA and OGD/R condition. * p
    Anti Bax, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 80/100, based on 0 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti bax/product/Merck KGaA
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti bax - by Bioz Stars, 2020-04
    80/100 stars
      Buy from Supplier

    91
    Merck KGaA mouse anti bax
    Reduction of apoptotic molecules following the inhibition of ASK1. (A) The levels of anti-apoptotic <t>(Bcl-2)</t> and pro-apoptotic <t>(Bax)</t> proteins were detected with Western blot analyses. (B) The relative optical density indicated that neuronal Bcl-2 expression was reduced after incubating the cells with EC-CM from the H/R group. After incubation with NQDI-1-treated EC-CM, the Bcl-2 level was increased ( n = 3). (C) The increased Bax level that was observed after incubating the cells with H/R-injured EC-CM was reduced after NQDI-1 treatment ( n = 3-4). (D) The level of apoptotic (caspase-3) protein was measured by Western blot analyses. (E) The graph showed the cleaved caspase-3 was increased in H/R-injured EC-CM, whereas ASK1-inhibited group exhibited lower cleaved caspase-3 level in neuronal cells ( n = 3). (F) Cell viability was detected with viability assays. The quantitative analyses show that neuronal cells were significantly dead after incubation with H/R-injured EC-CM; however, the cell viability was increased by treatment with ASK1 inhibitor ( n = 7). β-actin was used as an internal control. [Bars represent mean ± SEM, n = 3–4. Relative optical density (OD) of Bcl-2: H/R, 0. 79 ± 0.04; H/R+NQDI-1, 1.10 ± 0.05. OD of Bax: H/R, 1.27 ± 0.08; H/R+NQDI-1, 1.01 ± 0.04. OD of cleaved caspase-3: H/R, 1.17 ± 0.04; H/R+NQDI-1, 0.92 ± 0.05. * p
    Mouse Anti Bax, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti bax/product/Merck KGaA
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    mouse anti bax - by Bioz Stars, 2020-04
    91/100 stars
      Buy from Supplier

    98
    Merck KGaA gapdh
    The <t>p53</t> synthesis efficiency under different stress conditions in NIH3T3 cells. The cells were exposed for 16 hours to doxorubicin (0.1 μM), tunicamycin (1.2 μM) and thapsigargin (0.1 μM), or to an equivalent volume of DMSO, and then harvested. Endogenous p53 protein level was determined by western blot (a) or flow cytometry (b) using monoclonal 1C12 antibody. <t>GAPDH</t> was used as a loading control. The RT-PCR analysis of p53 mRNA and β-actin mRNA (as a control) was also conducted. For RT-PCR analysis, the samples were prepared using total RNA extracted from mouse fibroblasts treated with DMSO as a control, tunicamycin, thapsigargin and doxorubicin.
    Gapdh, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 98/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gapdh/product/Merck KGaA
    Average 98 stars, based on 54 article reviews
    Price from $9.99 to $1999.99
    gapdh - by Bioz Stars, 2020-04
    98/100 stars
      Buy from Supplier

    Image Search Results


    Inhibition of endothelial cell conditioned medium (EC-CM) induced pro-apoptotic molecule expression by isoflurane. To examine the effect of pretreated isoflurane on neuronal cell survival and death under culture with EC-CM, we performed western blot analysis using Bcl-2 and Bax. (B) The Bcl-2 protein was significantly upregulated in neuronal cells incubated with EC-CM of isoflurane, RAP, or isoflurane RAP treated groups in the tPA and OGD/R condition. (C) In contrast to Bcl-2, the protein levels of Bax was significantly downregulated in neuronal cells incubated with EC-CM of isoflurane, RAP or isoflurane + RAP-treated groups in the tPA and OGD/R condition. (E) Bcl-2 protein levels were significantly increased in neuronal cells incubated with EC-CM of isoflurane, MG-132, or isoflurane + MG-132-treated groups in the tPA and OGD/R condition. (F) Bax was significantly decreased in neuronal cells incubated with EC-CM of isoflurane, MG-132 and isoflurane + MG-132-treated groups in the tPA and OGD/R condition. * p

    Journal: International Journal of Medical Sciences

    Article Title: Isoflurane preconditioning inhibits the effects of tissue-type plasminogen activator on brain endothelial cell in an in vitro model of ischemic stroke

    doi: 10.7150/ijms.18037

    Figure Lengend Snippet: Inhibition of endothelial cell conditioned medium (EC-CM) induced pro-apoptotic molecule expression by isoflurane. To examine the effect of pretreated isoflurane on neuronal cell survival and death under culture with EC-CM, we performed western blot analysis using Bcl-2 and Bax. (B) The Bcl-2 protein was significantly upregulated in neuronal cells incubated with EC-CM of isoflurane, RAP, or isoflurane RAP treated groups in the tPA and OGD/R condition. (C) In contrast to Bcl-2, the protein levels of Bax was significantly downregulated in neuronal cells incubated with EC-CM of isoflurane, RAP or isoflurane + RAP-treated groups in the tPA and OGD/R condition. (E) Bcl-2 protein levels were significantly increased in neuronal cells incubated with EC-CM of isoflurane, MG-132, or isoflurane + MG-132-treated groups in the tPA and OGD/R condition. (F) Bax was significantly decreased in neuronal cells incubated with EC-CM of isoflurane, MG-132 and isoflurane + MG-132-treated groups in the tPA and OGD/R condition. * p

    Article Snippet: After blocking with 5% bovine serum albumin (BSA), membranes were incubated with the anti-LRP (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-NF-κB p65 (1:500, Santa Cruz Biotechnology), anti-Cox-2 (1:500, Santa Cruz Biotechnology), anti-Bax (1:1000, Merck Millipore, Bedford, MA, USA), anti-Bcl-2 (1:1000, Abcam, Cambridge, UK) primary antibodies respectively.

    Techniques: Inhibition, Expressing, Western Blot, Incubation

    Bufalin alters the apoptosis-related protein levels in A375.S2 cells. A total of 1 × 10 6 A375.S2 cells in 6-well plates were treated with 450 nM bufalin for 0, 6, 12, 24, and 48 h. Cells were harvested from each sample and associated proteins were measured by using SDS-PAGE and Western blotting as described in Section 2 . The protein levels of caspase-3, caspase-8, and caspase-9 (a), cytochrome c , AIF, Endo G, Bcl-X, and Bax (b), and Fas, FasL, and GRP78 (c) expressions were examined. β -Actin was used as an internal control to ensure equal loading.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Triggering Apoptotic Death of Human Malignant Melanoma A375.S2 Cells by Bufalin: Involvement of Caspase Cascade-Dependent and Independent Mitochondrial Signaling Pathways

    doi: 10.1155/2012/591241

    Figure Lengend Snippet: Bufalin alters the apoptosis-related protein levels in A375.S2 cells. A total of 1 × 10 6 A375.S2 cells in 6-well plates were treated with 450 nM bufalin for 0, 6, 12, 24, and 48 h. Cells were harvested from each sample and associated proteins were measured by using SDS-PAGE and Western blotting as described in Section 2 . The protein levels of caspase-3, caspase-8, and caspase-9 (a), cytochrome c , AIF, Endo G, Bcl-X, and Bax (b), and Fas, FasL, and GRP78 (c) expressions were examined. β -Actin was used as an internal control to ensure equal loading.

    Article Snippet: Anti-endonuclease G (Endo G) (Cat. AB3639) and anti-Bax (Cat. 04-434) were bought from Merck Millipore (Billerica,MA,USA).

    Techniques: SDS Page, Western Blot

    Reduction of apoptotic molecules following the inhibition of ASK1. (A) The levels of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) proteins were detected with Western blot analyses. (B) The relative optical density indicated that neuronal Bcl-2 expression was reduced after incubating the cells with EC-CM from the H/R group. After incubation with NQDI-1-treated EC-CM, the Bcl-2 level was increased ( n = 3). (C) The increased Bax level that was observed after incubating the cells with H/R-injured EC-CM was reduced after NQDI-1 treatment ( n = 3-4). (D) The level of apoptotic (caspase-3) protein was measured by Western blot analyses. (E) The graph showed the cleaved caspase-3 was increased in H/R-injured EC-CM, whereas ASK1-inhibited group exhibited lower cleaved caspase-3 level in neuronal cells ( n = 3). (F) Cell viability was detected with viability assays. The quantitative analyses show that neuronal cells were significantly dead after incubation with H/R-injured EC-CM; however, the cell viability was increased by treatment with ASK1 inhibitor ( n = 7). β-actin was used as an internal control. [Bars represent mean ± SEM, n = 3–4. Relative optical density (OD) of Bcl-2: H/R, 0. 79 ± 0.04; H/R+NQDI-1, 1.10 ± 0.05. OD of Bax: H/R, 1.27 ± 0.08; H/R+NQDI-1, 1.01 ± 0.04. OD of cleaved caspase-3: H/R, 1.17 ± 0.04; H/R+NQDI-1, 0.92 ± 0.05. * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Blockade of Apoptosis Signal-Regulating Kinase 1 Attenuates Matrix Metalloproteinase 9 Activity in Brain Endothelial Cells and the Subsequent Apoptosis in Neurons after Ischemic Injury

    doi: 10.3389/fncel.2016.00213

    Figure Lengend Snippet: Reduction of apoptotic molecules following the inhibition of ASK1. (A) The levels of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) proteins were detected with Western blot analyses. (B) The relative optical density indicated that neuronal Bcl-2 expression was reduced after incubating the cells with EC-CM from the H/R group. After incubation with NQDI-1-treated EC-CM, the Bcl-2 level was increased ( n = 3). (C) The increased Bax level that was observed after incubating the cells with H/R-injured EC-CM was reduced after NQDI-1 treatment ( n = 3-4). (D) The level of apoptotic (caspase-3) protein was measured by Western blot analyses. (E) The graph showed the cleaved caspase-3 was increased in H/R-injured EC-CM, whereas ASK1-inhibited group exhibited lower cleaved caspase-3 level in neuronal cells ( n = 3). (F) Cell viability was detected with viability assays. The quantitative analyses show that neuronal cells were significantly dead after incubation with H/R-injured EC-CM; however, the cell viability was increased by treatment with ASK1 inhibitor ( n = 7). β-actin was used as an internal control. [Bars represent mean ± SEM, n = 3–4. Relative optical density (OD) of Bcl-2: H/R, 0. 79 ± 0.04; H/R+NQDI-1, 1.10 ± 0.05. OD of Bax: H/R, 1.27 ± 0.08; H/R+NQDI-1, 1.01 ± 0.04. OD of cleaved caspase-3: H/R, 1.17 ± 0.04; H/R+NQDI-1, 0.92 ± 0.05. * p

    Article Snippet: Membranes were blocked with 5% BSA and incubated in the rabbit anti-phosphatidylinositol 3-kinase (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-pAkt (1:1000, Cell signaling, Danvers, MA, USA), rabbit anti-Akt (1:1000, Cell signaling), rabbit anti-nuclear factor erythroid 2 [NF-E2]-related factor 2 (1:1000, Santa Cruz Biotechnology), rabbit anti- HO-1 (1:1000, Abcam, Cambridge, UK), rabbit anti-Bcl-2 (1:1000, Abcam), mouse anti-Bax (1:1000, Merck Millipore), goat anti-cyclooxygenase-2 (1:500, Santa Cruz Biotechnology), rabbit anti-cleaved caspase-3 (1:1000, Santa Cruz Biotechnology), and rabbit anti-caspase-3 (1:1000, Merck Millipore) which is diluted in 2% BSA.

    Techniques: Inhibition, Western Blot, Expressing, Incubation

    The p53 synthesis efficiency under different stress conditions in NIH3T3 cells. The cells were exposed for 16 hours to doxorubicin (0.1 μM), tunicamycin (1.2 μM) and thapsigargin (0.1 μM), or to an equivalent volume of DMSO, and then harvested. Endogenous p53 protein level was determined by western blot (a) or flow cytometry (b) using monoclonal 1C12 antibody. GAPDH was used as a loading control. The RT-PCR analysis of p53 mRNA and β-actin mRNA (as a control) was also conducted. For RT-PCR analysis, the samples were prepared using total RNA extracted from mouse fibroblasts treated with DMSO as a control, tunicamycin, thapsigargin and doxorubicin.

    Journal: RNA Biology

    Article Title: Length and secondary structure of the 5′ non-coding regions of mouse p53 mRNA transcripts - mouse as a model organism for p53 gene expression studies

    doi: 10.1080/15476286.2018.1556084

    Figure Lengend Snippet: The p53 synthesis efficiency under different stress conditions in NIH3T3 cells. The cells were exposed for 16 hours to doxorubicin (0.1 μM), tunicamycin (1.2 μM) and thapsigargin (0.1 μM), or to an equivalent volume of DMSO, and then harvested. Endogenous p53 protein level was determined by western blot (a) or flow cytometry (b) using monoclonal 1C12 antibody. GAPDH was used as a loading control. The RT-PCR analysis of p53 mRNA and β-actin mRNA (as a control) was also conducted. For RT-PCR analysis, the samples were prepared using total RNA extracted from mouse fibroblasts treated with DMSO as a control, tunicamycin, thapsigargin and doxorubicin.

    Article Snippet: The blot was probed with mouse monoclonal antibodies against p53 (1C12; Cell Signaling), and GAPDH (Anti-Glyceraldehyde-3-Phosphate Dehydrogenase, clone 6C5ʹ, Merck Millipore).

    Techniques: Western Blot, Flow Cytometry, Cytometry, Reverse Transcription Polymerase Chain Reaction