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Anti phb2

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Anti-PHB2 is a primary antibody that specifically detects prohibitin 2 (PHB2), a protein involved in mitochondrial function and cellular stress response. This antibody can be used for various applications, including Western blotting and immunohistochemistry.

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Lab products found in correlation

Anti bcl 2, by Cell Signaling Technology (1 mentions) Anti p akt, by Cell Signaling Technology (1 mentions) Anti lc3a b, by Cell Signaling Technology (1 mentions) Anti p62, by Cell Signaling Technology (1 mentions) Pcdna3.1 cloning vector, by Genewiz (1 mentions) Anti mtor, by Cell Signaling Technology (1 mentions) Anti p mtor, by Cell Signaling Technology (1 mentions) Anti pink1, by Cell Signaling Technology (1 mentions) Anti caspase 7, by Cell Signaling Technology (1 mentions) Anti β actin, by Merck Group (1 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Agilent Technologies (1 mentions) Genesnap, by Syngene (1 mentions) Anti eif4e, by Cell Signaling Technology (1 mentions) Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions) Anti phospho eif4e, by Cell Signaling Technology (1 mentions) Mouse anti parp, by Santa Cruz Biotechnology (1 mentions) Complete mini, by Roche (1 mentions) Digital imaging system, by LI COR (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions) Anti ndufa9, by Abcam (1 mentions)

4 protocols using anti phb2

1

Plasmid Constructs and Antibodies

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PDI-Flag, PHB2-Flag plasmids constructed with pcDNA3.1(+) cloning vector were purchased from GENEWIZ company (Suzhou, China). LC3-His, PDI-Myc, plasmids constructed with pET-28a (+) cloning vector was purchased from Genomeditech Company (Shanghai, China). The antibodies were purchased from Proteintech (IL, USA) as follows: anti-PDI (11245-1-AP), anti-PEPK (20582-1-AP), anti-ATF6 (24169-1-AP), anti-TOM20 (11802-1-AP), anti-TOM40 (18409-1-AP), anti-Calnexin (10427-1-AP), anti-IRE1 (27528-1-AP), anti-PARP (13371-1-AP), anti-AKT (60203-2-Ig), anti-Actin (66009-1-Ig), anti-LC3 (14600-1-AP). The antibodies were purchased from Cell Signaling (MA, USA) as follows: anti-P62 (#39749), anti-PHB2 (#14085), anti-PINK1 (#6946), anti-mTOR (#2972), anti-p-mTOR (#5536), anti-Bcl-2 (#4223), anti-Caspase-7 (#9492), anti-p-AKT (#4060), anti-LC3A/B (#4108). The antibodies were purchased from Santa (CA, USA) as follows: anti-GRP78 (sc-13539), and anti-PHB2 (sc-133094). The antibodies were purchased from Beyotime (Shanghai, China) as follows: anti-FLAG (AF0036), anti-HIS (AF5060), and anti-MYC (AF0033).
Wang R., Shang Y., Chen B., Xu F., Zhang J., Zhang Z., Zhao X., Wan X., Xu A., Wu L, & Zhao G. (2022). Protein disulfide isomerase blocks the interaction of LC3II-PHB2 and promotes mTOR signaling to regulate autophagy and radio/chemo-sensitivity. Cell Death & Disease, 13(10), 851.
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2

Western Blot Analysis of Protein Signaling

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Proteins were obtained from whole cell lysates using lysis buffer (20 mM Tris–HCl [pH 7.5], 1% NP-40, 10% glycerol, and 150 mM NaCl supplemented with 1% protease and phosphatase inhibitor mixtures) (Sigma) as previously described [28 (link)]. The primary Abs used were: mouse anti-Akt1/2, rabbit anti-phospho-Ser473Akt, rabbit anti-Erk1/2 or anti-phospho-Erk1/2, anti-Mnk1, anti-4E-BP1, anti-AIF, anti-PHB1, anti-PHB2, anti-phospho(Ser/Thr)-Akt substrate, anti-eIF4E, anti-Phospho-eIF4E, anti-C-Raf and anti-phospho-C-Raf (Ser289/296/301) (Cell signaling Technology), mouse anti-PARP, rabbit anti-Bcl-2 and anti-c-Myc (Santa Cruz Biotechnology), and anti-βactin (Sigma). Horseradish peroxidase (HRP)–conjugated secondary antibodies (1:1000) were from DakoCytomation. Blotted proteins were detected and quantified using the Immobilon Western Chemiluminescent HRP Substrate (Millipore) and a bioimaging system (Genesnap; Syngene).
Bentayeb H., Aitamer M., Petit B., Dubanet L., Elderwish S., Désaubry L., de Gramont A., Raymond E., Olivrie A., Abraham J., Jauberteau M.O, & Troutaud D. (2019). Prohibitin (PHB) expression is associated with aggressiveness in DLBCL and flavagline-mediated inhibition of cytoplasmic PHB functions induces anti-tumor effects. Journal of Experimental & Clinical Cancer Research : CR, 38, 450.
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3

Immunoblotting in Whole Cells and Mitochondria

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For immunoblotting in whole cell lysates, cells were homogenized in RIPA (50mM Tris HCl, pH of 7.4, 150mM NaCl, 1.0% (v/v) NP-40, 0.5% (w/v) sodium deoxycholate, and 1.0mM EDTA, 0.1% (w/v) SDS) buffer plus 1% protease inhibitors (05892791001, Complete Mini; Roche, Mannheim, Germany). Equal amounts of proteins (25 – 50 μg) were separated by SDS-PAGE and transferred to PVDF membranes. For immunoblotting in mitochondria, equal amounts of proteins from crude mitochondrial preps were lysed in RIPA with 1% protease inhibitor for 30 minutes on ice before SDS-PAGE. Antibodies used were anti-PHB antibody (1:2000 dilution, clone # II-14-10, Thermo Scientific), anti-PHB2 (1:1000, Cell Signaling Technology), anti-HSP60 (1:1000, source), anti-SLP-2 (1:1000, ProteinTech), anti-β-actin (1:3000, Cell Signaling Technology). Representative subunits of CI were detected with anti-NDUFA9 (1:1000, Abcam) and anti-NDUFVI (1:500, Santa Cruz) antibodies. Complex III was detected with an anti-Core 2 subunit antibody (1:1000, Abcam). Protein bands were detected using a LI-COR Digital Imaging system and band intensities were quantified with Image Studio software.
Anderson C.J., Kahl A., Qian L., Stepanova A., Starkov A., Manfredi G., Iadecola C, & Zhou P. (2018). Prohibitin is a positive modulator of mitochondrial function in PC12 cells under oxidative stress. Journal of neurochemistry, 146(3), 235-250.
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Western Blot Validation of Proteomics

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To validate proteomics results, western blot analysis was performed. After the transfer of target proteins from SDS-PAGE, blocked membranes were incubated with Millipore anti-ROCK2 (Rho-associated protein kinase 2) (#07-1458), anti- IQGAP1 (Ras GTPase-activating-like protein 1) (#abt186), anti-PHB1 (#abn293), anti-PHB2 (#mabc953); Cell Signaling anti-GAPDH (#2118), anti-ILK1 (Integrin-linked protein kinase 1) (#3862), and Anti-Histone H3 (#4499) overnight at 4 °C. The membrane was rinsed with 1x TBS buffer prior to incubation with secondary antibody for 1 h. Blots were developed using ECL buffer (GE Healthcare) and the Geliance 600 imaging system (PerkinElmer).
Choi S., Bhagwat A.M., Al Mismar R., Goswami N., Ben Hamidane H., Sun L, & Graumann J. (2018). Proteomic profiling of human cancer pseudopodia for the identification of anti-metastatic drug candidates. Scientific Reports, 8, 5858.
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