β actin 66009 1 ig

Manufactured by Proteintech

Sourced in United States, China

β-actin (66009-1-Ig) is a primary antibody targeting the β-actin protein. β-actin is a widely expressed cytoskeletal protein that plays a fundamental role in various cellular processes.

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Lab products found in correlation

Gapdh 60004 1 ig, by Proteintech (4 mentions) Ripa lysis buffer, by Beyotime (2 mentions) Gsk3β, by Cell Signaling Technology (2 mentions) Gapdh 10494 1 ap, by Proteintech (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Hrp conjugated goat anti mouse igg, by Proteintech (2 mentions) Ab181873, by Abcam (2 mentions) Tom40 18409 1 ap, by Proteintech (2 mentions) Ab170108, by Abcam (2 mentions) Bca kit, by Sangon (2 mentions) Nitrocellulose membranes, by BBI Lifesciences (2 mentions) Protease and phosphatase inhibitor, by CWBIO (1 mentions) Immobilon ecl substrate, by Merck Group (1 mentions) Ripa lysis buffer, by CWBIO (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Image studio software, by LI COR (1 mentions) Chemidoc xr, by Bio-Rad (1 mentions) Beyoecl plus kit, by Beyotime (1 mentions) Ab41684, by Abcam (1 mentions) Ab69506, by Abcam (1 mentions)

Close spelling variants of this product

β-actin (1408 citations) Anti-β-actin (66009-1-Ig) (24 citations) Anti-β-actin antibody (66009-1-Ig) (6 citations) Mouse anti-β-actin (66009-1-Ig, (5 citations) -actin (66009-1-Ig, (4 citations) 66009-I-Ig (2 citations) β-Actin (cat no. 66009-1-Ig) (2 citations) Mouse anti-β-actin monoclonal antibody (66009-1-Ig; (2 citations)

45 protocols using β actin 66009 1 ig

Protein Expression Analysis by Western Blot

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Proteins from cells were extracted using radioimmunoprecipitation assay (RIPA) lysis buffer (CWBIO, China) with protease and phosphatase inhibitors (CWBIO, China). Identical quantities of proteins were electrophoresed by SDS-PAGE, transferred onto PVDF membranes, blocked with 5% non-fat milk for 1 h, and incubated with primary antibodies specific for Bax, Bcl-2, Caspase-3 (#2772, #2872, and #9662) (1 : 1000, Cell Signaling Technology, USA), and β-actin (66009-1-Ig, 1 : 5000, Proteintech, USA) at 4°C overnight, followed by incubation with anti-rabbit/mouse HRP-conjugated secondary antibodies (cw0103s, cw0102s) (1 : 2000, CWBIO, China) at room temperature for 1 h. Signals were detected by Immobilon ECL substrate (Millipore, Germany).

Ye Y., Jiang S., Zhang C., Cheng Y., Zhong H., Du T., Xu W., Azziz R., Zhang H, & Zhao X. (2020). Environmental Pollutant Benzo[a]pyrene Induces Recurrent Pregnancy Loss through Promoting Apoptosis and Suppressing Migration of Extravillous Trophoblast. BioMed Research International, 2020, 8983494.

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Breast Cancer Cell Line Maintenance and Reagents

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MCF-7 and MDA-MB-231 originated from ATCC and were maintained in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Life Technologies, Carlsbad, CA, USA) at 37 °C in 5% CO₂. Reagents used in this study were as follows: MTT (M2128), DAPI (D9542), Flubendazole (HY-B0294), Paclitaxel (HY-B0015), MMP-2 (87809, CST), E-cadherin (14472, CST), β-actin (66009-1-Ig, Proteintech, Rosemont, IL, USA), N-Cadherin (13116, CST), PI3K (4255, CST), p-PI3K (17366, CST), AKT (9272, CST), p-AKT (4060S, CST), mTOR (2972, CST), p-mTOR (5536, CST), HIF1α (36169, CST) p62 (88588, CST), LC3 (4108, CST), ATG5 (12994, CST).

Zhou Y., Liao M., Li Z., Ye J., Wu L., Mou Y., Fu L, & Zhen Y. (2023). Flubendazole Enhances the Inhibitory Effect of Paclitaxel via HIF1α/PI3K/AKT Signaling Pathways in Breast Cancer. International Journal of Molecular Sciences, 24(20), 15121.

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Western Blot Protein Analysis

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Western blotting analysis was performed as described previously.²³ Primary antibodies were used as follows: p‐mTOR(Ser2448) (D9C2; CST), p‐mTOR(Ser2448) (D9C2; CST), β‐actin (66009–1‐Ig, Proteintech), mTOR (D9C2; CST), p‐p70S6K(T389) (108D2;CST), p70S6K (49D7; CST), p‐AKT(S473) (D9E; CST), AKT(pan) (C67E7; CST). The quantification of the band intensities was done using ImageStudio software (LI‐COR Inc.).

Funasaki S., Mehanna S., Ma W., Nishizawa H., Kamikubo Y., Sugiyama H., Ikeda S., Motoshima T., Hasumi H., Linehan W.M., Schmidt L.S., Ricketts C., Suda T., Oike Y., Kamba T, & Baba M. (2022). Targeting chemoresistance in Xp11.2 translocation renal cell carcinoma using a novel polyamide–chlorambucil conjugate. Cancer Science, 113(7), 2352-2367.

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Quantitative Western Blot Analysis of Neuroinflammatory Markers

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Protein from each right hemisphere sample was extracted, quantified, and denatured at 95°C in 5X loading buffer for 10 min. Western blot was performed as previously described (17 (link)). The following primary antibodies were used: Fibrin (ogen) (ab34269, Abcam), P-selectin (60322-1-Ig, Proteintech), PDGFRβ (ab69506, Abcam), CypA (ab41684, Abcam), N-cadherin (22018-1-AP, Proteintech), phospho-NF-κB p65 subunit (p-p65, ab86299, Abcam), MMP-9 (10375-2-AP, Proteintech), and β-actin (66009-1-Ig, Proteintech). The bands were visualized using a BeyoECL Plus kit (P0018; Beyotime) and photographed by a chemiluminescence imaging system (ChemiDoc XRS+; Bio-Rad, Hercules, CA, USA). Band densities were quantified by a blinded observer using ImageJ software.

Pang J., Wu Y., Peng J., Yang P., Chen L, & Jiang Y. (2021). Association of Pericyte Loss With Microthrombosis After Subarachnoid Hemorrhage in ApoE-Deficient Mice. Frontiers in Neurology, 12, 726520.

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Protein Expression Analysis by Western Blot

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Cells were lysed on ice using RIPA lysis buffer (Beyotime Biotechnology, Shanghai, China), and then the protein extracts were separated by SDS-PAGE. The primary antibodies used in this study included β-actin (66009-1-Ig, Proteintech, Chicago, IL, USA), COPS5 (6895, CST), CTR1 (20868-1-AP, Proteintech, Chicago, IL, USA), GSTP1 (15902-1-AP, Proteintech, Chicago, IL, USA), GSTT1 (15838-1-AP, Proteintech, Chicago, IL, USA), TOPO1 (sc-271285, Santa Cruz, CA, USA). The secondary antibodies contained HRP-conjugated Goat Anti-Rabbit IgG (SA00001-2, Proteintech, Chicago, IL, USA) and HRP-conjugated Goat Anti-Mouse IgG (SA00001-1, Proteintech, Chicago, IL, USA).

Zhang H., Yan T., Zhong A., Guo L, & Lu R. (2022). COPS5 Conferred the Platinum Resistance in Epithelial Ovarian Cancer. Current Issues in Molecular Biology, 44(9), 3948-3958.

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Alzheimer's Disease Protein Profiling

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The following antibodies were used: PARP16 (ARP33751) was obtained from Aviva (Aviva Systems Biology, USA); Aβ 6E10 (800304) was obtained from BioLegend (BioLegend, Inc. USA); sAPPβ (10321) and sAPPα (11088) were obtained from IBL (Immuno-Biological Laboratories Co., Ltd., Japan); p-IRE1α (PA1-16927) was obtained from ThermoFisher (Thermo Fisher Scientific Inc., USA); XBP-1 (ab37152), and Bcl-2 (ab196495) were obtained Abcam (Abcam); APP (2450), p-PERK (3179), p-eIF2α (3597), Bax (2772), Caspase 3 (9662), and p53 (2524) were purchased from CST (Cell Signaling Technology, Inc., USA); COX-2 (A3560), IL-1β (A1112) and IL-6 (A0286) were obtained from ABclonal (ABclonal Technology Co.,Ltd. China); iNOS (18985-1-AP), BIP (11587-1-AP), Calnexin (10427-2-AP), β-tubulin (10094-1-AP), β-Actin (66009-1-Ig), and GAPDH (60004-1-Ig) were purchased from Proteintech (Proteintech Group, Inc., USA). Actinomycin D was purchased from Linkchem (Shanghai LinkChem Technology Co., Ltd. China). MG132 was purchased from Meilunbio (Dalian, China). 3-Methyladenine (3-MA) and tauroursodeoxycholic acid (TUDCA) were obtained from MCE (MedChemExpress, USA). Aβ_1-42 (A9810) was purchased from Sigma-Aldrich (Sigma-Aldrich, USA).

Wang J., Cheng Q., Zhang Y., Hong C., Liu J., Liu X, & Chang J. (2023). PARP16-Mediated Stabilization of Amyloid Precursor Protein mRNA Exacerbates Alzheimer’s Disease Pathogenesis. Aging and Disease, 14(4), 1458-1471.

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Immunohistochemical Analysis of Signaling Pathways

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Antibodies against β-actin (#66009-1-Ig), IL-1β (#16806-1-AP), IL-6 (#66146-1-Ig), and Fibronectin (#15613-1-AP), as well as goat anti-rabbit (#SA00001-1) and goat anti-mouse (#SA00001-15) secondary antibodies were purchased from Proteintech (Rosemont, IL, USA). Antibodies against TSC1 (#6935), p-AMPK Thr-172 (#2535), AMPK (#5832), p-Akt Ser-473 (#4060), p-Akt Thr-308 (#13038), Akt (#2920), PCNA (#2586), Caspase-3 and Cleaved caspase-3 (#9664), BAD (#9239), BAX (#5023), F4/80 (#30325), CD3 (#99940), CD4 (#25229), and α-SMA (#19245) were purchased from Cell Signaling Technology (Danvers, MA, USA). Rabbit anti-Ki-67 (#15580) was purchased from ABCAM. Fluorescein-labeled LTL (#FL-1321) and Dylight 594 anti-rabbit IgG (#DI-1594) were purchased from Vector Labs. Peroxidase marker and DAB + Substrate (#5007) were purchased from Dako.

Fang Y., Li F., Qi C., Mao X., Wang F., Zhao Z., Chen J.K., Zhang Z, & Wu H. (2020). Metformin effectively treats Tsc1 deletion-caused kidney pathology by upregulating AMPK phosphorylation. Cell Death Discovery, 6, 52.

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Immunoblotting Antibody Detection Protocol

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Immunoblotting was performed as described previously 30 (link), 31 (link). Anti-HLA-F (14670-1-AP), -HK1 (19662-1-AP), -HK2 (22029-1-AP), -PKM1 (15821-1-AP), -PFKM (55028-1-AP), -Myc (16286-1-AP), and -β-actin (66009-1-Ig) antibodies were purchased from Proteintech.

Chen X., Sun N., Li R., Sang X., Li X., Zhao J., Han J., Yang J, & Ikezoe T. (2021). Targeting HLA-F suppresses the proliferation of glioma cells via a reduction in hexokinase 2-dependent glycolysis. International Journal of Biological Sciences, 17(5), 1263-1276.

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Comprehensive Antibody Validation Protocol

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Western blot was performed according to the previous method [18 (link)]. Primary antibodies for c-Fos (sc-166940), p-c-Fos (#sc-81485), GFP-tag (sc-9996), V5-tag (sc-81594), and p62 (sc-28359) were purchased from Santa Cruz Biotechnology (Santa Cruz; CA, USA). Primary antibodies for p-ERK1/2 (#4370), LC3 (#4180), CD44 (#37259), K63-linkage ubiquitin (#5621), and β-tubulin (#2148) were obtained from Cell Signaling Technology (CST; Danvers, MA, USA). Primary antibodies for NANOG (A3232), OCT4 (#A7920), SOX2 (#A0561), KLF4 (A13673), Caspase 3 (#A2156), Flag-tag (AE092) were sourced from Abclonal (Wuhan, China). Primary antibodies for TMPO (#14651-1-AP), Lamin A/C (#10298-1-AP), ERK1/2 (#67170-1-Ig), and β-actin (#66009-1-Ig) were acquired from Proteintech (Wuhan, China). Primary antibody for K48-linkage ubiquitin (#ab140601) was purchased from Abcam (Cambridge, UK).

Gui Y., Qian X., Ding Y., Chen Q., Fangyu Y.e., Ye Y., Hou Y., Yu J, & Zhao L. (2024). c-Fos regulated by TMPO/ERK axis promotes 5-FU resistance via inducing NANOG transcription in colon cancer. Cell Death & Disease, 15(1), 61.

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Antibody Protocols for Protein Detection

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The specific antibody against cGAS (26416–1-AP), NMHC-IIA (11128–1-AP), NMHC-IIA (60233–1-Ig), Flag (DYKDDDDK) tag (66008–4-Ig), HA tag (66006–2-Ig), glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 60004–1-Ig), PARP1 (13371–1-AP), or β-actin (66009–1-Ig) was purchased from Proteintech (Wuhan, China). The specific antibody against phospho-TBK1/NAK (Ser172) or TBK1/NAK (D1B4) was from Cell Signaling Technology (Danvers, USA). The antibody against PRV US9 was purchased from Developmental Studies Hybridoma Bank (Antibody Registry ID: AB_1553789, Bethesda, USA). Alexa Fluor 488-labeled goat anti-rabbit immunoglobulin (IgG, H + L; A0423) and Alexa Fluor 647-labeled goat anti-mouse IgG (H + L; A0473) were purchased from Beyotime (Shanghai, China).

Liu Y., Qin Y., Yang B., Zheng H., Qiao S., Luo Z, & Li R. (2024). Pseudorabies virus usurps non-muscle myosin heavy chain IIA to dampen viral DNA recognition by cGAS for antagonism of host antiviral innate immunity. Journal of Virology, 98(5), e00483-24.

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