Myricetin (#HY-15097) was purchased from MedChemExpress (New Jersey, United States), dissolved in dimethyl sulfoxide (DMSO, #D8370, Solarbio, Beijing, China) at a concentration of 100 mM, and stored at −20 °C. MG132 (#HY-13259) and BafA1 (#HY-100558) were also bought from MedChemExpress and dissolved in DMSO. All antibodies were as follows: anti-Bcl-2 (#12789-1-AP), Ki-67 (#27309-1-AP), Stat3 (#10253-2-AP), LC3 (#14600-1-AP), P62 (#18420-1-AP), CyclinB1 (#55004-1-AP), CyclinD1 (#26939-1-AP), GAPDH (#10494-1-AP), and peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H + L) (#SA00001-2) (Proteintech Group, Chicago, IL, United States); MARCH1 (#bs-9335R, Bioss, Beijing, China); p-Stat3 (#ab32143), p38 MAPK (#ab170099) (abcam, Cambridge, United Kingdom); p-p38 MAPK (#11581, Singalway Antibody, Maryland, United States); and MARCH1 (#YT2642, Immunoway, Newark, United States).
Anti bcl 2
Manufactured by Proteintech
Sourced in United States, China, United Kingdom
Anti-Bcl-2 is a laboratory equipment product designed for research purposes. It is an antibody that specifically binds to the Bcl-2 protein, which is involved in the regulation of apoptosis (programmed cell death). This product can be used in various research applications, such as Western blotting, immunohistochemistry, and flow cytometry, to detect and quantify the expression of Bcl-2 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti bax, by Proteintech (89 mentions)
Anti caspase 3, by Proteintech (22 mentions)
Anti gapdh, by Proteintech (21 mentions)
Anti β actin, by Proteintech (20 mentions)
Pvdf membrane, by Merck Group (15 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (12 mentions)
Anti cleaved caspase 3, by Proteintech (10 mentions)
Bca protein assay kit, by Beyotime (10 mentions)
Anti lc3, by Proteintech (9 mentions)
Anti caspase 9, by Proteintech (8 mentions)
Anti akt, by Cell Signaling Technology (8 mentions)
Anti beclin 1, by Proteintech (8 mentions)
Anti e cadherin, by Proteintech (8 mentions)
Anti p53, by Proteintech (7 mentions)
Anti caspase 3, by Cell Signaling Technology (7 mentions)
Anti akt, by Proteintech (7 mentions)
Anti cytochrome c, by Proteintech (7 mentions)
Anti p62, by Proteintech (5 mentions)
Anti vimentin, by Proteintech (5 mentions)
Ripa lysis buffer, by Beyotime (5 mentions)
126 protocols using anti bcl 2
1
Myricetin modulates autophagy pathways
2
Comprehensive Western Blot Protocol
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Western blot was performed as previously described 44 (link). The antibodies included anti-Oct4 (Bioss, Beijing, China, bs-0830R, 1:1000), anti-Nanog (Proteintech, Wuhan, China, 14295-1-AP, 1:1000), anti-CD44 (Proteintech, 15675-1-AP, 1:1000), anti-Sox2 (Proteintech, 11064-1-AP, 1:1000), anti-γ-H2AX (Abcam, Cambridge, UK, ab26350, 1:1000), anti-DLG2 (Affinity Biosciences, Suzhou, China, DF3995, 1:1000), anti-p-Yap1 (CST, Boston MA, USA, 4911S, 1:1000), anti-Yap1 (Proteintech, 13584-1-AP, 1:1000), anti-Bax (CST, B8429, 1:1000), anti-Bcl2 (Proteintech, 60178-1-Ig, 1:1000), anti-β-tubulin (Sigma-Aldrich, T5201, 1:1000), anti-Lamin B1 (Santa, Dallas TX, USA, sc-365962, 1:1000), anti-TEAD1 (ABclonal, Wuhan, China, A13366, 1:1000), anti-β-actin (Santa, sc-8432, 1:4000), CD63 (Proteintech, 25682-1-AP, 1:1000), and TSG101 (Proteintech, 28283-1-AP, 1:1000).
3
Evaluating Inflammatory Signaling and Renal Function
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TNF-α, IL-1β, and IL-6 ELISA Kits were purchased from Biolegend (San Diego, CA,
USA). rmMFG-E8 was purchased from R&D Systems (Minneapolis, MN, USA).
Antibodies used for western blot were as follows: anti-Bcl-2 (Proteintech,
Rosemont, IL, USA), anti-Bax (Proteintech, Rosemont, IL, USA), anti-ß-actin
(Proteintech, Rosemont, IL, USA), and anti-histone H3 (Proteintech, Rosemont,
IL, USA). A NF-κB signal pathway kit was used to determine NF-κB pathway
activation (Cell Signaling Technology, Beverly, MA, USA). Blood urea nitrogen
(BUN) and serum creatinine (Cre) detection kits were purchased from the
Institute of Jiancheng Bioengineering (Nanjing, China). All other reagents were
of analytical grade.
USA). rmMFG-E8 was purchased from R&D Systems (Minneapolis, MN, USA).
Antibodies used for western blot were as follows: anti-Bcl-2 (Proteintech,
Rosemont, IL, USA), anti-Bax (Proteintech, Rosemont, IL, USA), anti-ß-actin
(Proteintech, Rosemont, IL, USA), and anti-histone H3 (Proteintech, Rosemont,
IL, USA). A NF-κB signal pathway kit was used to determine NF-κB pathway
activation (Cell Signaling Technology, Beverly, MA, USA). Blood urea nitrogen
(BUN) and serum creatinine (Cre) detection kits were purchased from the
Institute of Jiancheng Bioengineering (Nanjing, China). All other reagents were
of analytical grade.
4
Protein Expression Analysis by Western Blot
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Cells were harvested after the indicated treatments and dissolve RIPA buffer; protein concentrations quantification by bicinchoninic acid detection kit (Pierce, USA). Protein samples were Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) separation under denaturing conditions and transferred to a nitrocellulose filter (NC) membrane. The membrane was then incubated with the following primary antibodies: anti-β-tubulin (CST), anti-GAPDH (CST), anti-CDC20 (Proteintech), anti-BCL-2 (CST), anti-BAX (CST), anti-p21 (Proteintech), anti-P53 (Proteintech), anti-CCNB1 (CST), anti-CCND1 (CST).
5
Western Blot Analysis of Renal Proteins
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Renal tissues were homogenized in RIPA lysis buffer (Sigma, St. Louis, MO, USA). Protein concentrations of the homogenates were measured by the BCA protein assay kit (Beyotime Institute of Biotechnology, Shanghai, China). The extracted proteins (50 μg) were separated on SDS-PAGE gel and transferred to a PVDF-nitrocellulose membrane. After blocking, the membranes were incubated with primary antibodies to detect the target proteins. Anti-extracellular signal-regulated protein kinase 1/2 (ERK1/2), anti-phospho (p)-ERK1/2 (Thr202/Tyr204), anti-c-Jun N-terminal kinase (JNK), anti-p-JNK (Thr183/Tyr185), anti-p38, anti-p-p38 (Thr180/Tyr182), anti-p50, anti-p65, and anti-p-p65 (Ser536) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-Bax, anti-Bcl-2, anti-Caspase-3, anti-Cleaved Caspase-3, and anti-β-actin antibodies were purchased from ProteinTech (Chicago, IL, USA). The horseradish peroxidase-conjugated secondary antibody was purchased from Cell Signaling Technology. The reaction was visualized using an enhanced chemiluminescence system (Thermo Fisher Scientific, Rockford, IL, USA). The bands were quantified by densitometry using ImageJ software.
6
Western Blot Analysis of Apoptosis Markers
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Cells were washed twice in PBS and lysed in RIPA buffer (Solarbio, China) with 1 mM phenylmethanesulfonyl fluoride (PMSF). A BCA protein kit was used to quantify protein concentrations (Beyotime, China). Total proteins (30 μg) were separated on 15% SDS-PAGE gels and transferred to polyvinylidene difluoride (PVDF) membranes (Beyotime, China). The membranes were then blocked in TBST with 3% non-fat milk (Sangon Biotech, China) at 25°C for 2 h and incubated with anti-survivin (1:1,000), anti-caspase-3 (1:1,000), anti-Bcl-2 (1:3,000), anti-P-gp (1:3,000), and anti-β-actin (1:5,000) primary antibodies (Proteintech, USA) overnight at 4°C. Next, the membranes were incubated with the appropriate HRP-conjugated secondary antibodies (1:10,000) (Proteintech, USA) at 25°C for 1 h. Protein bands were visualized by ECL chemiluminescence kit (Sangon Biotech, China), and the gray values of the protein bands were analyzed by Image J.
7
Western Blot Analysis of Apoptosis Markers
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Cells were lysed with 1 mL ice-cold lysis buffer (1% triton x-100, 20 mm Tris-Hcl, ph 7.5, 150 mm NaCl, 10 mm naf, 1 mm Na3vo4, 10 mm PMSF, 1 mm benzaminidine, 5 mg/mL aprotinin, 3 mg/mL pepstatin, 5 mg/mL leupeptin). The cell lysate was centrifuged at 15,000 rpm, 4 °C for 20 min. Next, the protein concentration was measured using BCA protein assay (Beyotime, Beijing, China). The proteins were heated at 100 °C for 10 min, and then equal amount of protein per group was resolved on 10% SDS-PAGE gel. Next, the proteins were transferred to nitrocellulose (NC) membranes. Thereafter, the membranes were blocked with 5% skim milk for 2 hrs at room temperature and incubated with the following primary antibodies: rabbit anti-Caspase-3 (1:500 dilution; Proteintech, America), anti-Caspase-9 (1:300 dilution; Proteintech, Wuhan, China), anti-Bcl-2 (1:1000 dilution; Proteintech, America), anti-Bax (1:6000 dilution; Proteintech, Wuhan, China), anti-Cytc (1:5000 dilution; ABCAM, Britain), and mouse anti-β-actin (1:2000 dilution; Cell Signaling Technology, Shanghai, China). Subsequently, the NC membranes were washed and incubated with the corresponding goat anti-mouse or anti-rabbit IgG-HRP (Bioworld Technology, China) at room temperature for 2 h. Finally, the immunoreactive signals were detected with SuperSignal ECL (Pierce, Rockford, IL, USA).
8
Immunohistochemical Analysis of Apoptosis Markers
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Tumors excised from mice were fixed in 10% formalin, embedded in paraffin, and cut into 4-mm sections. Deparaffinized tumor sections were treated with 3% H2O2 for 10 min to block endogenous peroxidases and incubated with 5% blocking serum (goat serum) at room temperature for 30 min. After blocking, the slides were incubated with polyclonal rabbit anti-Caspase-3 (1:200 dilution; Proteintech, America), anti-Caspase-9 (1:100 dilution; Proteintech, Wuhan, China), anti-Bcl-2 (1:200 dilution; Proteintech, America), anti-Bax (1:200 dilution; Proteintech, Wuhan, China), and anti-Cytc (1:250 dilution; ABCAM, Britain) overnight at 4°C. The sections were then incubated for 1 h with a biotin labeled secondary antibody, followed by avidin-peroxidase reagent and 3,3ʹ-diaminobenzidine (DAB; Fuzhou, China) substrate. After hematoxylin counterstaining and dehydration, the sections were sealed with cover slips. Finally, the stained setions were observed under a microscope.
9
Western Blotting of Protein Levels
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Protein levels in mouse tissues or cells were determined by western blotting using the following antibodies: anti‐TPPP2 (Abcam, 121215; 1:1000), anti‐green fluorescent protein (GFP) (EnoGene, E12‐026‐4; 1:3000), anti‐β‐ACTIN (Merck Millipore, MAB1501; 1:7500), anti‐Caspase3 (Proteintech, 19677‐1‐AP; 1:500), anti‐Bax (Proteintech, 50599‐2‐Ig; 1:500), anti‐Bcl‐2 (Proteintech, 12789‐1‐AP; 1:500) and anti‐Eef1b (Proteintech, 10483‐1‐P; 1:1000).
10
Western Blot Analysis of Apoptosis Regulators
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Total protein was extracted from the cells using ice-cold RIPA lysis buffer and the protein concentration was measured using the BCA Protein Assay Kit. 30 µg total proteins were applied on 10% SDS-PAGE and transferred onto 0.45 mm PVDF membranes (Millipore, United States). The membranes were blocked with 5% non-fat milk for 2 h at room temperature, and then incubated with primary antibodies overnight at 4°C. After washing with TBST buffer three times, the membranes were incubated with secondary HRP-conjugated antibodies for 1 h at room temperature. Capture specific bands using the ECL detection system. The primary antibodies were anti-B4GALT1 (Abcam, ab121326, 1:1000), anti-Bcl-2(Proteintech, 12789-1-AP, 1:1000), anti-Bax (Proteintech, 50,599-2-lg, 1:1000), anti-C-caspase (Proteintech, 19677-1-AP, 1:1000) and anti-β-actin (Proteintech, 20536-1-AP, 1:1000).
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