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Ab129069

Manufactured by Abcam
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Sourced in United States, United Kingdom, Germany

Ab129069 is a laboratory product manufactured by Abcam. It is a primary antibody that can be used for research purposes. The core function of this product is to detect the specified target in samples.

Automatically generated - may contain errors

Lab products found in correlation

Ab654, by Abcam (2 mentions) Ab110410, by Abcam (2 mentions) Ab84622, by Abcam (2 mentions) Ab60128, by Abcam (2 mentions) Nbp1 54154, by Novus Biologicals (2 mentions) Nbp1 40422, by Novus Biologicals (2 mentions) Ab10694, by Abcam (2 mentions) Ab131263, by Abcam (2 mentions) Ab178423, by Abcam (2 mentions) Ab2302, by Merck Group (2 mentions) Ab179463, by Abcam (1 mentions) Ab32028, by Abcam (1 mentions) Ab155996, by Abcam (1 mentions) Ab134187, by Abcam (1 mentions) Pierce protein g magnetic beads, by Thermo Fisher Scientific (1 mentions) Turbofect transfection reagent, by Thermo Fisher Scientific (1 mentions) Cholesterol, by Merck Group (1 mentions) Ab81283, by Abcam (1 mentions) Nitrocellulose membrane, by Cytiva (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions)

7 protocols using ab129069

1

Multiparameter Immunoblotting Analysis

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Donkey anti Goat IR Dye 680RD (1: 10,000), 926-68074, Donkey anti Mouse IRDye 680RD (1: 10,000), Goat anti Rabbit IRDye 800CW (1: 10,000), 926-32211 (All Licor Biosciences Ltd), Goat anti Aconitase 2, Ab99467 (1: 10000), Rabbit anti Aconitase 2, Ab129069 (1: 50,000), Goat anti Aconitase 1, Ab10694 (1: 500), Complex II Ab Cocktail, Ab110410 (1: 250), Rabbit anti Glut4, Ab654 (1: 2,000), Rabbit anti SDHB, Ab84622 (1: 1,000), Rabbit anti TST, Ab60128, (1: 2,000)(All Abcam), Rabbit anti SDHB (1: 7,000), NBP1-54154, Rabbit anti SOD2 (1; 2,000), NBP1-40422, Goat anti SOD1 (1: 10,000), all Novus Biologicals, NB100-60944, Rabbit polyclonal anti-TST antibody, GTX 114858 (1: 1,000), Source Biochemicals.
Morton N.M., Beltram J., Carter R.N., Michailidou Z., Gorjanc G., Fadden C.M., Barrios-Llerena M.E., Rodriguez-Cuenca S., Gibbins M.T., Aird R.E., Moreno-Navarrete J.M., Munger S.C., Svenson K.L., Gastaldello A., Ramage L., Naredo G., Zeyda M., Wang Z.V., Howie A.F., Saari A., Sipilä P., Stulnig T.M., Gudnason V., Kenyon C.J., Seckl J.R., Walker B.R., Webster S.P., Dunbar D.R., Churchill G.A., Vidal-Puig A., Fernandez-Real J.M., Emilsson V, & Horvat S. (2016). Genetic identification of thiosulfate sulfurtransferase as an adipocyte-expressed anti-diabetic target in mice selected for leanness. Nature medicine, 22(7), 771-779.
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2

Mitochondrial Enzyme Profiling in Cardiomyocytes

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Cardiomyocytes were prepared and treated as above. Cells were lysed with RIPA buffer and 20 μg of protein was loaded onto 10% gels. Membranes were probed for succinate dehydrogenase (ab178423; Abcam), pyruvate dehydrogenase (ab131263; Abcam), aconitase (ab129069; Abcam), and GAPDH (AB2302, 1:5000; Millipore). Densitometric readings of band intensities were obtained using ImageJ software.
Dukinfield M., Maniati E., Reynolds L.E., Aubdool A., Baliga R.S., D'Amico G., Maiques O., Wang J., Bedi KC J.r., Margulies K.B., Sanz‐Moreno V., Hobbs A, & Hodivala‐Dilke K. (2019). Repurposing an anti‐cancer agent for the treatment of hypertrophic heart disease. The Journal of Pathology, 249(4), 523-535.
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3

Protein Analysis of Cellular Pathways

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High-glucose Dulbecco's modified Eagle's medium (H-DMEM) was obtained from Basalmedia Technologies (Shanghai, China) and fetal bovine serum (FBS) was obtained from Bailing Biotechnology (Lanzhou, China). TurboFect™ transfection reagent (R0531) and Pierce™ Protein G Magnetic Beads were from Thermo Fisher Scientific (Waltham, NY, USA). Cholesterol was obtained from Sigma-Aldrich (St. Louis, MO, USA). The primary antibodies, rabbit monoclonal anti-AKT1 (ab81283), anti-AKT-1/2/3 (ab179463), anti-mTOR/p-mTOR (ab32028, ab109268), anti-HO-1 (ab52947), anti-Aconitase2 (Aco2) (ab129069), anti-lactate dehydrogenase (LDH) (ab134187), anti-pyruvate dehydrogenase E-1β (PDHE-1β) (ab155996), anti-nuclear factor erythroid 2-like 2 (Nrf2) (ab62352) and anti-EBP (ab135745) were purchased from Abcam (Cambridge, MA, USA). Anti-histone H3.1 was obtained from Abmart. Anti-malic dehydrogenase (MDH2) (#8610) was obtained from Cell Signaling Technology. The anti-rabbit HRP-labeling and the anti-rabbit DyLight™ 488- and anti-mouse DyLight™ 633-labeled secondary antibodies were from KPL.
Jin X., Xu Z., Cao J., Yan R., Xu R., Ran R., Ma Y., Cai W., Fan R., Zhang Y., Zhou X, & Li Y. (2017). HO-1/EBP interaction alleviates cholesterol-induced hypoxia through the activation of the AKT and Nrf2/mTOR pathways and inhibition of carbohydrate metabolism in cardiomyocytes. International Journal of Molecular Medicine, 39(6), 1409-1420.
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4

Western Blot Analysis of Mitochondrial Proteins

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Less than 100 mg of snap‐frozen tissue was lysed in RIPA buffer supplemented with protease inhibitor cocktail (Merck‐Sigma, Gillingham, UK). Tissue was homogenised using a Polytron tissue homogeniser for 30 s followed by centrifugation, for 15 min at 4 °C, to pellet tissue debris. Lysates were subjected to SDS‐PAGE and transferred to nitrocellulose membranes (Amersham Biosciences, Amersham, UK) for western blotting. Blots were probed for β3 integrin (antibody a kind gift from Barry Coller, Rockefeller University; 1:500), succinate dehydrogenase (ab178423, 1:1000; Abcam, Cambridge, UK), pyruvate dehydrogenase (ab131263, 1:1000; Abcam), aconitase (ab129069, 1:10 000; Abcam), and GAPDH (AB2302; Merck, Hoddeston, UK). Densitometric readings of band intensities were obtained using ImageJ software (NIH, USA).
Dukinfield M., Maniati E., Reynolds L.E., Aubdool A., Baliga R.S., D'Amico G., Maiques O., Wang J., Bedi KC J.r., Margulies K.B., Sanz‐Moreno V., Hobbs A, & Hodivala‐Dilke K. (2019). Repurposing an anti‐cancer agent for the treatment of hypertrophic heart disease. The Journal of Pathology, 249(4), 523-535.
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5

Western Blot Antibody Panel for Iron Regulation

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The following antibodies were used for Western blot analysis: anti-EIF3B (Bethyl A301-761A) at 1:1000; anti-FTL (Abcam, ab69090) at 1:800; anti-FTH (Santa Cruz, sc-25617) at 1:400; anti-IRP1 (Abbexa, abx004618) at 1:400; anti-IRP2 (Abcam, ab129069) at 1:800; and anti-b-Actin (Abcam, ab8227) at 1:1000, anti-Ferroportin (Novus, NBP 1–2150255) at 1:300.
Pulos-Holmes M.C., Srole D.N., Juarez M.G., Lee A.S., McSwiggen D.T., Ingolia N.T, & Cate J.H. (2019). Repression of ferritin light chain translation by human eIF3. eLife, 8, e48193.
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6

Multiparameter Immunoblotting Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Donkey anti Goat IR Dye 680RD (1: 10,000), 926-68074, Donkey anti Mouse IRDye 680RD (1: 10,000), Goat anti Rabbit IRDye 800CW (1: 10,000), 926-32211 (All Licor Biosciences Ltd), Goat anti Aconitase 2, Ab99467 (1: 10000), Rabbit anti Aconitase 2, Ab129069 (1: 50,000), Goat anti Aconitase 1, Ab10694 (1: 500), Complex II Ab Cocktail, Ab110410 (1: 250), Rabbit anti Glut4, Ab654 (1: 2,000), Rabbit anti SDHB, Ab84622 (1: 1,000), Rabbit anti TST, Ab60128, (1: 2,000)(All Abcam), Rabbit anti SDHB (1: 7,000), NBP1-54154, Rabbit anti SOD2 (1; 2,000), NBP1-40422, Goat anti SOD1 (1: 10,000), all Novus Biologicals, NB100-60944, Rabbit polyclonal anti-TST antibody, GTX 114858 (1: 1,000), Source Biochemicals.
Morton N.M., Beltram J., Carter R.N., Michailidou Z., Gorjanc G., Fadden C.M., Barrios-Llerena M.E., Rodriguez-Cuenca S., Gibbins M.T., Aird R.E., Moreno-Navarrete J.M., Munger S.C., Svenson K.L., Gastaldello A., Ramage L., Naredo G., Zeyda M., Wang Z.V., Howie A.F., Saari A., Sipilä P., Stulnig T.M., Gudnason V., Kenyon C.J., Seckl J.R., Walker B.R., Webster S.P., Dunbar D.R., Churchill G.A., Vidal-Puig A., Fernandez-Real J.M., Emilsson V, & Horvat S. (2016). Genetic identification of thiosulfate sulfurtransferase as an adipocyte-expressed anti-diabetic target in mice selected for leanness. Nature medicine, 22(7), 771-779.
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7

Western Blot Analysis of Mitochondrial Proteins

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Fibroblasts were grown under standard conditions and lysed with RIPA buffer containing Complete Protease Inhibitor (Roche, Germany). For each sample, we harvested ~ 500,000 cells and loaded 30 µg of total protein on 10% polyacrylamide gels and resolved by one-dimensional discontinuous sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins were blotted on nitrocellulose membrane (Invitrogen GmbH, Karlsruhe, Germany) by using the iBlot 2 device (Invitrogen GmbH, Karlsruhe, Germany) for 7 min at 20 V. Proteins of interest were labeled with primary antibodies against ACO2 (anti-rabbit; Abcam: ab129069; dilution: 1:1000), β-Actin (anti-mouse; Cell signal: 37005; dilution: 1:5000), TOM20 (anti-rabbit; Santa Cruz: Sc-11415; dilution: 1:1000) and secondary antibodies goat anti-mouse IgG (Novex: A24524; dilution: 1:10,000) or goat anti-rabbit IgG (Novex: A24537; 1:5000), respectively. Protein bands were visualized with Amersham ECL Western Blotting Detection Reagent (GE healthcare, Freiburg, Germany) on the ODYSSEY chemiluminescence 2800 Fc (Li-COR, Lincoln, USA). Three independent samples per cell line (n = 3) were assessed. Image J software was used for relative signal quantification.
Neumann M.A., Grossmann D., Schimpf-Linzenbold S., Dayan D., Stingl K., Ben-Menachem R., Pines O., Massart F., Delcambre S., Ghelfi J., Bohler J., Strom T., Kessel A., Azem A., Schöls L., Grünewald A., Wissinger B, & Krüger R. (2020). Haploinsufficiency due to a novel ACO2 deletion causes mitochondrial dysfunction in fibroblasts from a patient with dominant optic nerve atrophy. Scientific Reports, 10, 16736.
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