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Ab71498

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Ab71498 is a laboratory equipment product. It is designed to perform a specific function in a laboratory setting. The core function of this product is to facilitate a particular experimental or analytical process. No further details about the intended use or specific applications of this product can be provided in an unbiased and factual manner.

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Lab products found in correlation

Ab56788, by Abcam (4 mentions) Ab57602, by Abcam (3 mentions) Ab184247, by Abcam (2 mentions) Ab131607, by Abcam (2 mentions) Ab104274, by Abcam (2 mentions) Ab56889, by Abcam (2 mentions) Ab90857, by Abcam (2 mentions) β actin, by Proteintech (2 mentions) Bcl 2 antibody, by Abcam (1 mentions) Hrp 60008, by Proteintech (1 mentions) Ab192306, by Abcam (1 mentions) Bax antibody, by Cell Signaling Technology (1 mentions) Kindlin 2 antibody, by Cell Signaling Technology (1 mentions) Chop antibody, by Cell Signaling Technology (1 mentions) Pdi antibody, by Cell Signaling Technology (1 mentions) Goat anti rabbit igg antibody, by Proteintech (1 mentions) Goat anti mouse igg antibody, by Proteintech (1 mentions) Sa00001 1, by Proteintech (1 mentions) Sa00001 2, by Proteintech (1 mentions) Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)

12 protocols using ab71498

1

Protein Extraction and Western Blot Analysis

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The protein extraction and Western blot analysis were performed as previous described.22 The antibody information was as following: kindlin‐2 antibody (13562s, Cell Signaling Technology), Bax antibody (14796, Cell Signaling Technology), Bcl‐2 antibody (ab194583, Abcam), GRP78 antibody (GRP78, 3183, Cell Signaling Technology), CHOP antibody (2895, Cell Signaling Technology), PDI antibody (3501, Cell Signaling Technology), Ero1‐Lα antibody (ab81959, Abcam), Drp‐1 antibody (ab184247, Abcam), Tfam antibody (ab131607, Abcam), ND3 (ab192306, Abcam), Mfn‐2 (9482, Cell Signaling Technology), Fis‐1 (ab71498, Abcam), ATPB (ab170947, Abcam), β actin Antibody (HRP‐60008, Proteintech), Goat anti‐rabbit IgG antibody (SA00001‐2, Proteintech) and Goat anti‐mouse IgG antibody (SA00001‐1, Proteintech). The Image J software was used for quantitative analysing, and relative protein levels were expressed as the intensity ratio of target protein and β actin.
Zhang J., Ren Y., Bi J., Wang M., Zhang L., Wang T., Wei S., Mou X., Lv Y, & Wu R. (2020). Involvement of kindlin‐2 in irisin’s protection against ischaemia reperfusion‐induced liver injury in high‐fat diet‐fed mice. Journal of Cellular and Molecular Medicine, 24(22), 13081-13092.
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2

Mitochondrial Dynamics Protein Analysis

Cited 2 times
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The primary antibodies used in the present study were as follows [30 (link)]: Drp1 (1:1000, Abcam, #ab56788), Fis1 (1:1000, Abcam, #ab71498), Opa1 (1:1000, Abcam, #ab42364), Mfn1 (1:1000, Abcam, #ab57602), Mfn2 (1:1000, Abcam, #ab56889), Mff (1:1000, Cell Signaling Technology, #86668), Bcl2 (1:1000, Cell Signaling Technology, #3498), Bax (1:1000, Cell Signaling Technology, #2772), caspase9 (1:1000, Cell Signaling Technology, #9504), survivin (1:1000, Cell Signaling Technology, #2808), p53 (1:1000, Cell Signaling Technology, #9282), complex III subunit core (CIII-core2, 1:1000, Invitrogen, #459220), complex II (CII-30, 1:1000, Abcam, #ab110410), complex IV subunit II (CIV-II, 1:1000, Abcam, #ab110268), Tom20 (1:1,000, Abcam, #ab186735) [31 (link)].
Zhao S., Li P., Wang P., Yang J., Song P., Zhang D, & Zhou G. (2019). Nurr1 Promotes Lung Cancer Apoptosis Via Enhancing Mitochondrial Stress and p53-Drp1 Pathway. Open Life Sciences, 14, 262-274.
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3

Mitochondrial Dynamics and Apoptosis Analysis

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Cell pellets were lysed in RIPA buffer containing 1× HaltTM protease inhibitor cocktail (78437, Thermo) and supernatants collected after 10 min centrifugation at 12000 × g for SDS-PAGE. Total proteins were transferred to PVDF membranes and blotted with the antibodies against FIS1 (ab71498, Abcam), MFN1 (ab104274, Abcam), β-Actin (ab8227, Abcam), Caspase-3 (9665, Cell Signaling), Caspase-8 (sc-5263, Santa Cruz), Caspase-9 (sc-8355, Santa Cruz), HSP70 (TA309356, Origene) and Total OXPHOS Human WB Antibody Cocktail (ab110411, Abcam).
Gonzalez-Ibanez A.M., Ruiz L.M., Jensen E., Echeverria C.A., Romero V., Stiles L., Shirihai O.S, & Elorza A.A. (2020). Erythroid Differentiation and Heme Biosynthesis Are Dependent on a Shift in the Balance of Mitochondrial Fusion and Fission Dynamics. Frontiers in Cell and Developmental Biology, 8, 592035.
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4

Western Blot Analysis of Mitochondrial Dynamics Proteins

Cited 2 times
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Proteins (60–80 µg) were loaded on a 10–15% sodium dodecyl sulfate-polyacrylamide gel. After electrophoresis, proteins were transferred to a PVDF membrane. Membrane were blocked with 5% nonfat dried milk (in TBST) for 2 h at room temperature and then incubated overnight at 4 °C with primary antibodies. The membrane was subsequently washed with TBST (5 min × 3) and incubated with horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology) for 1 h at room temperature [33] (link). After washing with TBST (5 min × 3), bands were detected by enhanced chemiluminescence substrate (Applygen). The antibodies used in the present were as follows: Drp1 (1:1000, Abcam, #ab56788), p-Drp1 (Ser616) (1:1000, Cell Signaling Technology, #3455), p-Drp1 (Ser637) (1:1000, Abcam, #ab193216), Fis1 (1:1000, Abcam, #ab71498), Mfn2 (1:1000, Abcam, #ab56889), Mfn1 (1:1000, Abcam, #ab57602), Mff (1:1000, Cell Signaling Technology, #86668), F-actin (1:1000, Abcam, #ab205), G-actin (1:1000, Abcam, #ab200046), p-eNOS (Ser1117) (1:1000, Abcam, #ab184154), ICAM1 (1:1000, Abcam, #ab119871), VCAM1 (1:1000, Abcam, #ab134047), AMPK (1:1000, Abcam, #ab131512), p-AMPK (1:1000, Abcam, #ab23875) [34] (link).
Zhou H., Wang S., Zhu P., Hu S., Chen Y, & Ren J. (2017). Empagliflozin rescues diabetic myocardial microvascular injury via AMPK-mediated inhibition of mitochondrial fission. Redox Biology, 15, 335-346.
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5

Comprehensive Protein Extraction and Western Blot Analysis

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Total protein was extracted using a cell lysis buffer containing 50 mM Tris (pH 7.4), 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS (Beyotime, Shanghai, China) supplemented with each protease and phosphatase inhibitor cocktail tablet (Roche Diagnostics GmbH, Mannheim, Germany) per 10 ml solution. Western blot was performed as previously described 57 (link). Primary antibodies against IRF1 (sc-514544x) and β-actin (sc-47778) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The antibodies against P62 (5114), p-mTOR (5536), t-mTOR (2983), p-p70S6K (9234), p-4EBP1 (2855), PINK1 (6946) and Parkin (4211) were obtained from Cell Signaling Technology (CST, Danvers, MA, USA). The antibodies against DRP1 (ab56788), MFN1 (ab57602), MFN2 (ab50838), OPA1 (ab90857), FIS1 (ab71498), AhR (ab2769), COXIV (ab14744), PGC1α (ab54481) and TFAM (ab131607) were purchased from Abcam (Cambridge, MA, USA). LC3 antibody (L7543) was purchased from Sigma-Aldrich.
Huang Y., Zhou J., Wang S., Xiong J., Chen Y., Liu Y., Xiao T., Li Y., He T., Li Y., Bi X., Yang K., Han W., Qiao Y., Yu Y, & Zhao J. (2020). Indoxyl sulfate induces intestinal barrier injury through IRF1-DRP1 axis-mediated mitophagy impairment. Theranostics, 10(16), 7384-7400.
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6

Mitochondrial Dysfunction Evaluation Protocol

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The antibodies against p62 (1 : 1000, 5114S), DRP1 (1 : 1000, 8570), phospho‐DRP1 (1 : 1000, 3455), p38 (1 : 1000, 8690), phospho‐p38 (1 : 1000, 4511), PINK1 (1 : 1000, 6949), Parkin (1 : 1000, 4211), OPA1 (1 : 1000, 67589), MFF (1 : 1000, 84580), Mfn1 (1 : 1000, 14739), Mfn2 (1 : 1000, 9482), and cleaved caspase‐3 (1 : 1000, 9664) was purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against HSF1‐S326 (1 : 1000, ab76760), FIS1 (1 : 500, ab71498), FTL (1 : 1000, ab47369), FTH (1 : 1000, ab47369), and FPN1 (1 : 1000, ab85370) were purchased from Abcam (Cambridge, MA, USA). The antibody against TOMM20 was purchased from Santa Cruz Biotechnology (Delaware, CA, USA). The antibody against HSF1 (1 : 1000, ABE1044) was obtained from Millipore (Bedford, MA, USA). Antibodies against TFR1 (1 : 1000, 10084‐2‐AP), Lamin A/C (1 : 500, 10298‐1‐AP), Bcl‐2 (1 : 1000, 12789‐1‐AP), Bax (1 : 1000, 50599‐2‐Ig), Nrf2 (1 : 1000, 16396‐1‐AP), NQO1 (1 : 1000, 11451‐1‐AP), GCLC (1 : 1000, 12601‐1‐AP), and β‐actin (1 : 1000, 66009‐1‐Ig) were purchased from ProteinTech (Wuhan, China). Erastin (S7242), celastrol (S1290), PD98059 (S1177), Z‐VAD‐FMK (S7023), Ferrostatin‐1 (S1177), KRIBB11 (S8402), and NAC (S1623) were purchased from Selleck Chemicals (Houston, TX, USA). DFO (D9533) and FAC (RES20400‐A7) were purchased from Sigma‐Aldrich (St. Louis, MO, USA).
Liu M., Fan Y., Li D., Han B., Meng Y., Chen F., Liu T., Song Z., Han Y., Huang L., Chang Y., Cao P., Nakai A, & Tan K. (2021). Ferroptosis inducer erastin sensitizes NSCLC cells to celastrol through activation of the ROS–mitochondrial fission–mitophagy axis. Molecular Oncology, 15(8), 2084-2105.
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7

Mitochondrial Dynamics Regulation Assay

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Oligomycin B (ab143424), antimycin A (ab141904), and antibodies against total OXPHOS (ab110413), Mfn1 (ab104274), Mfn2 (ab56889), DRP1 (ab56788), ubiquitin (Abcam, ab140601), FIS1 (ab71498), and OPA1 (ab90857) were obtained from Abcam (Cambridge, MA, USA). Antibodies to TOM20 (#42406), LC3A/B (#4108), P-DRP1 (Ser637, #4867), and β-Actin (#3700) were purchased from Cell Signaling Technology (Beverly, MA, USA). Mul1 (16133-1-AP) and Parkin (14060-1-AP) antibodies were purchased from Proteintech (Wuhan, Hubei, China). Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP, C2920), rotenone (R8875), and Mdivi-1 (M0199) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ginsenoside CK was purchased from Yuanye Bio-Technology (HPLC ≥ 98%, Shanghai, China).
Huang Q., Li J., Chen J., Zhang Z., Xu P., Qi H., Chen Z., Liu J., Lu J., Shi M., Zhang Y., Ma Y., Zhao D, & Li X. (2022). Ginsenoside compound K protects against cerebral ischemia/reperfusion injury via Mul1/Mfn2-mediated mitochondrial dynamics and bioenergy. Journal of Ginseng Research, 47(3), 408-419.
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8

Mitochondrial Dynamics in Cell Death

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Glucose (G8270), Mannitol (78513), CQ (C6628), and STZ (S0130) were purchased from Sigma. The siRNAs for Pink1 and Parkin, along with control siRNA and siRNA transfection reagent (human, sc-29528), were purchased from Ribobio Co., Ltd. Antibodies used for immunoblotting including rabbit anti-Parkin (ab15954); rabbit anti-Pink1 (ab23707) and rabbit anti-Fis1 (ab71498) were purchased from Abcam. Mouse anti-OPA1 (612606) was obtained from BD Biosciences. Mouse anti-Drp1 (14647); rabbit anti-LC3 (2775); rabbit anti-Tom20 (42406); rabbit anti-Bax (2772); rabbit anti-MFN2 (9482); rabbit anti-Cleaved Caspase-3 (9664); mouse anti-COXIV (11967) and rabbit anti-Cyt C (4272) were purchased from Cell Signaling Technology. Rabbit anti-β-actin (AC006) and rabbit anti-Bcl-2 (A2212) were obtained from Abclonal. Goat anti-CD31 (AF3628) was purchased from R&D Systems. Donkey Anti-Rabbit DyLight® 550 and Donkey anti-Goat Alexa Fluor 647 secondary antibodies for immunofluorescence were obtained from Abcam and Life Technologies, respectively.
Zhu W., Yuan Y., Liao G., Li L., Liu J., Chen Y., Zhang J., Cheng J, & Lu Y. (2018). Mesenchymal stem cells ameliorate hyperglycemia-induced endothelial injury through modulation of mitophagy. Cell Death & Disease, 9(8), 837.
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9

Immunoblotting of Mitochondrial Proteins

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Immunoblotting was performed as previously described [37 (link)]. Briefly, cells were lysed for 0.5 h at 4°C in a RIPA Buffer (R0278, Sigma) containing a protease inhibitor cocktail. Protein extracts were resolved through 8% SDS-polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membranes (BioRad, Berkeley, CA, USA), probed with antibody against human MFF (ab81127), DRP1 (ab56788), FIS1 (ab71498), TOM20(ab78547), BRCA1 (ab16780; Abcam), MFN1 (sc-50330), MFN2 (sc-50331), OPA1 (sc-393296), COXIV(sc-376731), cytochrome c (sc-13560) (Santa Cruz), β-actin (Proteintech, Chicago, IL, USA) or Tubulin (sc-53646) and then with a peroxidase-conjugated secondary antibody (Proteintech); they were visualized via chemiluminescence (GE, Fairfield, CT, USA).
Fan S., Liu B., Sun L., Lv X.B., Lin Z., Chen W., Chen W., Tang Q., Wang Y., Su Y., Jin S., Zhang D., Zhong J., Li Y., Wen B., Zhang Z., Yang P., Zhou B., Liang Q., Yu X., Zhu Y., Hu P., Chu J., Huang W., Feng Y., Peng H., Huang Q., Song E, & Li J. (2015). Mitochondrial fission determines cisplatin sensitivity in tongue squamous cell carcinoma through the BRCA1–miR-593-5p–MFF axis. Oncotarget, 6(17), 14885-14904.
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10

Molecular Profiling of Mitochondrial Dynamics

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Western blotting was performed as described previously using specific antibodies against α-SMA (ab7817, Abcam, 1:5000), TGFβ1 (sc-130348, Santa Cruz Biotechnology, 1:1500), Bnip3 (ab109362, Abcam, 1:7000), phospho-DRP1 (Ser616) (#4494, Cell Signaling Technology, 1:2000), Drp1 (#8570, Cell Signaling Technology, 1:2000), Fis1 (ab71498, Abcam, 1:2000), RORα (sc-6062, Santa Cruz Biotechnology, 1:2000), and actin (sc-1616, Santa Cruz Biotechnology, 1:2000). The ChIP assay was performed using anti-RORα (sc-6062, Santa Cruz Biotechnology), anti-p300 (sc-585, Santa Cruz Biotechnology), and anti-histone 3 (acetyl K27) (ab4729, Abcam) antibodies or a control IgG antibody (Santa Cruz Biotechnology), and specific primers (Supplementary Table)19 (link). Relative mRNA expression was determined by qRT–PCR using the ABI StepOnePlusTM Real-Time PCR system (Applied Biosystems, Foster City, CA) using specific primers (Supplementary Table). The mRNA expression of genes was calculated relative to controls using the 2−ΔΔCT method19 (link).
Kim H.J., Han Y.H., Na H., Kim J.Y., Kim T., Kim H.J., Shin C., Lee J.W, & Lee M.O. (2017). Liver-specific deletion of RORα aggravates diet-induced nonalcoholic steatohepatitis by inducing mitochondrial dysfunction. Scientific Reports, 7, 16041.
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