β actin ta 09

Manufactured by ZSGB-BIO

Sourced in China, United States

β-actin (TA-09) is a primary antibody that recognizes the beta-actin protein. β-actin is a widely expressed cytoskeletal protein that is involved in various cellular processes.

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β-actin (159 citations) TA-09 (62 citations) Anti-β-actin (TA-09) (4 citations) Anti-β-actin antibody (TA-09, (2 citations) Anti-β actin mAb (TA-09, (2 citations) Mouse anti-β-actin monoclonal antibody (TA-09) (2 citations)

9 protocols using β actin ta 09

Redox-Sensitive Protein Regulation Protocol

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The chemical (2S, 3S)-1,4-Bis-sulfanylbutane-2,3-diol (DTT, CAS No. 3483-12-3) was obtained from Sigma; it is a strong reductant with the chemical formula C₄H₁₀O₂S₂ and MW of 154.25 g/mol. Its reducibility is largely due to the conformational stability after oxidation (containing disulfide bond). In this experiment, 3.09 g DTT powder was completely dissolved in 20 mL 0.01 M sodium acetate to obtain 1 M DTT stock solution, which was packed and stored at −20 °C before use.
Specific antibody against Nrf1 was made in our own laboratory [25 (link)]. All nine distinct antibodies against Nrf2 (ab62352), GCLC (ab207777), GCLM (ab126704), HO-1 (ab52947), GPX1 (ab108427), XBP1 (AB109221), ATF4 (ab184909), ATF6 (ab227830) and P4HB (ab137180) were obtained from Abcam (Cambridge, UK). The first three antibodies against TALDO (D623398), GSR (D220726) and NQO1 (D26104) were purchased from Sangon Biotech (Shanghai, China). The second three antibodies against BIP (bs-1219R), Chop (bs-20669R) and pIRE1 (bs-16698R) were from Bioss (Beijing, China). The third three antibodies against PSMB5 (A1975), PSMB6 (A4054), PSMB7 (A14771) were from ABclonal (Wuhan, China). Lastly, three antibodies to p-eIF2α (#5199) were from Cell Signaling Technology (Boston, MA, USA), p-PERK (sc-32577) from Santa CruZ (Santacruz, CA, USA), and β-actin (TA-09) from ZSGB-BIO (Beijing, China), respectively.

Wufuer R., Fan Z., Yuan J., Zheng Z., Hu S., Sun G, & Zhang Y. (2022). Distinct Roles of Nrf1 and Nrf2 in Monitoring the Reductive Stress Response to Dithiothreitol (DTT). Antioxidants, 11(8), 1535.

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Deciphering Autophagy Pathways: A Comprehensive Protocol

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3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), bafilomycin A1 (Baf), wortmannin, monodansylcadaverine (MDC), 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), triphenyltetrazolium chloride (TTC) were purchased from Sigma. Antibodies against beclin-1 (3495), LC3B (#2775), p62(5114S) and Bax were obtained from Cell Signaling Technology, Danvers, Massachusetts, USA, anti-Bcl-2(SC-7382) was from Santa Cruz Biotechnology, USA. β-actin (TA-09) was from ZSGB-BIO, Beijing, China. Antimyoactin antibody, was obtained from Wuhan Boster Biological Technology, LTD., China. Lipofectamine RNAiMAX kit was purchased from Invitrogen Company.

Li X., Zeng Z., Li Q., Xu Q., Xie J., Hao H., Luo G., Liao W., Bin J., Huang X, & Liao Y. (2015). Inhibition of microRNA-497 ameliorates anoxia/reoxygenation injury in cardiomyocytes by suppressing cell apoptosis and enhancing autophagy. Oncotarget, 6(22), 18829-18844.

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Antibody Analysis of Cell Signaling

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The primary antibodies included AnxA6 (sc-166807) and N-cadherin (sc-31031) were ordered from Santa Cruz Company, SUMO1 (ET1606-53), ubiquitin (ET1609-21), AKT1(ET1609-47), p-AKT1^ser473(ET1607-73), vimentin (ET1610-39), E-cadherin (EM0502), twist (RT1635) and RHOU (ER1918-73) were ordered from HuaBio Company in China. AnxA6 (A5390) was ordered from ABclonal. The antibody β-actin (TA-09, Zsbio) was used to quantify the expression of housekeeping gene β-actin for comparison normalization.

Yang Y., Huang L., Zhang N., Deng Y.N., Cao X., Liang Y., Hou H., Luo Y., Yang Y., Li Q, & Liang S. (2024). SUMOylation of annexin A6 retards cell migration and tumor growth by suppressing RHOU/AKT1–involved EMT in hepatocellular carcinoma. Cell Communication and Signaling : CCS, 22, 206.

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Autophagy Regulation Signaling Pathway

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Alexa Fluor 488 Phalloidin (8878), caspase-3 (9665 s), caspase-9 (9502), light chain 3 (LC3)-I/II (2775), AKT (4685), phospho-AKT (4051), mTOR (2972), phospho-mTOR (5536), eukaryotic initiation factor 4E (eIF4E, 2067), phospho-eukaryotic initiation factor 4E (p-eIF4E, 9741), ribosomal protein S6 kinase (p70s6k) (9202), phospho-ribosomal protein S6 kinase, (p-p70s6k, 9206), serine/threonine kinase LKB1/STK11 (3047), phospho-LKB1 (3482), forkhead box O3a (FOXO3a, 2497), AMPK (2532), phospho-forkhead box O 3a (p-FOXO3a, 9465), UNC-51-like kinase-1 (ULK1, 4776), phospho-UNC-51-like kinase-1 (ULK1, 12,753), anti-rabbit secondary antibody (7054), and anti-mouse secondary antibody (7056) were purchased from Cell Signaling Technology, USA. Atg5 (ab78073) and phospho-AMPK (ab195946) were purchased from Abcam, UK. β-Actin (TA-09) was purchased from ZSGB-BIO, China. Other reagents were obtained from the following sources as required: delphinidin (Sigma, USA, 43725); 3-MA (Sigma, USA, M9281); BA1 (Cayman, USA, 88899–55-2); DMSO (Sigma, USA, D2650); and cell counting kit-8 (CCK-8) (Dojindo, Japan, CK04–20).

Chen J., Zhu Y., Zhang W., Peng X., Zhou J., Li F., Han B., Liu X., Ou Y, & Yu X. (2018). Delphinidin induced protective autophagy via mTOR pathway suppression and AMPK pathway activation in HER-2 positive breast cancer cells. BMC Cancer, 18, 342.

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Protein Expression Analysis in HUVECs

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The HUVECs were lyzed in 100 μL of RIPA buffer (50 mM Tris-cl pH 7.4, 150 mM NaCl, 1% NP40, and 0.25% Na-deoxycholate) containing 1x complete protease inhibitor cocktail (Roche Diagnostics, Indianapolis, IN) and 1x phosphatase inhibitor cocktail (Sigma, St. Louis, MO) on ice for 30 min. After centrifugation at 10000 ×g at 4°C for 15 min, the supernatant was collected and protein concentration was measured with Pierce Coomassie Plus (Bradford) Assay Kit (23236, ThermoFisher). 30 μg total protein was resolved in a 8% sodium dodecyl sulfate polyacrylamide gel and transferred onto a PVDF membrane, which was blocked in 5% nonfat milk in PBST (0.1% Tween 20 in PBS) at room temperature for 30 min followed by being incubated with specified first antibodies at 4°C overnight. Next day, the membranes were washed 3 times at 5 min each time with PBST and then incubated with proper horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) for 60 min at room temperature. The specific bands were detected with Pierce ECL Plus Substrate (ThermoFisher). The primary antibodies used were p-NF-κB p65 (ab86299), NF-κB p65 (ab16502), ICAM-1 (ab124759), and LOX-1 (ab60178) from Abcam (Cambridge, MA), p-p38MAPK (4511), and p38MAPK (8690) from Cell Signaling (Shanghai, China) and β-actin (TA-09) from ZSGB-Bio (Beijing, China).

Yao J., Zou Z., Wang X., Ji X, & Yang J. (2016). Pinoresinol Diglucoside Alleviates oxLDL-Induced Dysfunction in Human Umbilical Vein Endothelial Cells. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 3124519.

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TNF-α-induced necroptosis in MC3T3-E1 cells

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The MC3T3-E1 osteoblast cell line was provided by the Institute of Basic Medical Sciences (China). Cells were incubated in α-MEM medium (32571036; Gibco, USA) containing glutamine, 10% fetal bovine serum (10099-141; Gibco, Australia) and 1% penicillin/streptomycin (15140-122; Gibco, USA). Recombinant mouse TNF-α was purchased from Peprotech (AF-315-01A-20; USA). Nec-1 was purchased from MedChemExpress (4311-88-0; USA). Z-IETD-FMK, a specific inhibitor of caspase 8, was purchased from Selleck (S7314; USA). The primary antibodies used to detect p-MLKL (ab196436) and RIPK3 (ab56164) were purchased from Abcam (UK). The primary antibodies used to detect MLKL (37705S), cleaved caspase 3 (9661s), and caspase 3 (9662s) were obtained from Cell Signaling Technology (USA). β-actin (TA-09) and goat anti-mouse/rabbit IgG secondary antibodies were purchased from ZSGB-BIO (China).

Shi G., Jia P., Chen H., Bao L., Feng F, & Tang H. (2018). Necroptosis occurs in osteoblasts during tumor necrosis factor-α stimulation and caspase-8 inhibition. Brazilian Journal of Medical and Biological Research, 52(1), e7844.

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Prostate Cancer Cell Protein Analysis

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Prostate cancer cell lines stimulated in vitro as indicated above were lysed in the cell lysis buffer (Beyotime Biotechnology, China). The lysated were quantified with the bicinchoninic acid (BCA) kit (Beyotime Biotechnology, China). Then, the supernatants were subjected to 10% SDS-PAGE and then transferred to PVDF membranes (0.22 μm, Millipore) for detection of FOXO3a, catalase and β-actin. The blots were cut prior to hybridization with antibodies during blotting. Rabbit anti-FOXO3a (catalog no. 12829) was purchased from Cell Signaling Technology. Rabbit anti-catalase (catalog no. sc-50508) was from Santa Cruz Biotechnology. β-actin (TA-09) was from ZSGB-BIO (Beijing, China) and dye-labeled secondary antibody (IRDye800 or IRDye700) was from LI-COR. Band intensity was quantified using the Image Studio Software.

Tao Y., Liu S., Lu J., Fu S., Li L., Zhang J., Wang Z, & Hong M. (2022). FOXO3a-ROS pathway is involved in androgen-induced proliferation of prostate cancer cell. BMC Urology, 22, 70.

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Glioma Protein Expression Analysis

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Protein samples were extracted from glioma cell lines/glioma tissues. Protein samples were separated on 12.5% SDS-PAGE gels and transferred to PVDF membranes. The membranes were blocked with 5% skim milk and incubated with the primary antibody at 4 °C overnight. The next day, the membranes were incubated with fluorescent dye-conjugated secondary antibodies at room temperature for 2 h. The membranes were observed using a ChemiDoc XRS + Imaging System (Bio-Rad, USA). The following primary antibodies were utilized in the present study: P62 (18420-1-AP, Proteintech); MAP1LC3B (L7543, Sigma); CCND1 (AF0931, Affinity); CCNB1 (DF6786, Affinity); HAS3 (DF13055, Affinity); CD44 (A0340, Abclonal); and β-actin (TA-09, ZSGB-BIO).

Yan T., Chen X., Zhan H., Yao P., Wang N., Yang H., Zhang C., Wang K., Hu H., Li J., Sun J., Dong Y., Lu E., Zheng Z., Zhang R., Wang X., Ma J., Gao M., Ye J., Wang X., Teng L., Liu H, & Zhao S. (2021). Interfering with hyaluronic acid metabolism suppresses glioma cell proliferation by regulating autophagy. Cell Death & Disease, 12(5), 486.

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Necrostatin-1 Inhibits Necroptosis in Osteoblast and Breast Cancer Cells

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The MC3T3-E1 osteoblasts cell line and the 4T1 mouse breast cancer cell line were provided by the National Infrastructure of Cell Line Resource (China). Necrostatin-1 (Nec-1, 4311880, USA), the specific inhibitor of RIPK 1, was provided by MedChemExpress (USA). The specific inhibitor of caspase 8, named Z-IETD-FMK (S7314; USA), was purchased from Selleck (USA). These two reagents are soluble in DMSO (Methyl sulfoxide, 67-68-5, Sigma, USA), then were added medium to adjust concentration to 50 and 40 µM for subsequent experiments respectively. The primary antibodies of RIPK3 (ab56164) and p-MLKL (ab196436) were purchased from Abcam (UK), and the primary antibodies of cleaved caspase 3 (9661s), caspase 3 (9662s) and MLKL (37705S) were got from Cell Signaling Technology (USA). β-actin (TA-09) and goat anti-mouse/rabbit IgG secondary antibodies were obtained from ZSGB-BIO (China).
The MC3T3-E1 cells was incubated in α-MEM medium (32571036; Gibco, USA), which contained 1% penicillin/streptomycin (15140-122; Gibco, USA), 10% fetal bovine serum (10099-141; Gibco, Australia) and glutamine, in a constant temperature incubator with 5% CO₂ and 37 °C.

Ji X., Wang R., Tang H., Chen H., Bao L., Feng F, & Jia P. (2020). Necroptosis of osteoblasts was induced by breast cancer cells in vitro. Translational Cancer Research, 9(2), 500-507.

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