Laemmli buffer containing 4% SDS, 10% β-mercaptoethanol (M6250; Sigma-Aldrich), 20% glycerol (GI345; Melford), 0.004% blue bromophenol (A2331,0025; AppliChem), and 0.125 M Tris-HCl was added to protein lysates, followed by boiling at 95°C for 10 min. Samples were separated on SDS-PAGE and transferred to a nitrocellulose (Macherey-Nagel) or a polyvinylidene difluoride (Merck) membrane. The membranes were blocked with 5% milk (A0830; PanReac AppliChem) and probed with the following antibodies: anti–β-Actin (ab8227; Abcam), anti–β-Tubulin (ab15568; Abcam), anti-CBS (14787-1-AP; Proteintech), anti-CSE (12217-1-AP; Proteintech), anti-MPST (HPA001240; Atlas Antibodies), anti-MTCO1 (ab14705; Abcam), anti-Citrate Synthase (ab96600; Abcam), anti-SOD2 (13141; Cell Signaling), anti-TOM40L (ab236421; Abcam), anti-TIM50 (ab109436; Abcam), anti-HIF1α (sc-13515; Santa Cruz Biotechnology), anti-H3 (ab176842; Abcam), anti-V5 (Merck), anti-HA (H6908; Sigma-Aldrich), and anti-Biotin (HRP conjugate; 5571; Cell Signaling). Immunoblots were next incubated with a secondary antibody (anti-rabbit [AP132P; Merck] or anti-mouse [7076; Cell Signaling]) and visualized using the Western HRP substrate (Merck). ImageJ software was used to quantify the expression levels of the proteins.
Ab109436
Manufactured by Abcam
Ab109436 is an assay kit designed to quantify the level of protein expression in biological samples. The kit includes reagents and protocols necessary to perform the assay. Specific details about the target protein or intended applications are not provided.
Automatically generated - may contain errors
Lab products found in correlation
Ab14705, by Abcam (1 mentions)
Ab96600, by Abcam (1 mentions)
Polyvinylidene difluoride, by Merck Group (1 mentions)
Ab176842, by Abcam (1 mentions)
M6250, by Merck Group (1 mentions)
Sc 13515, by Santa Cruz Biotechnology (1 mentions)
Ab15568, by Abcam (1 mentions)
H6908, by Merck Group (1 mentions)
A0830, by ITW Reagents (1 mentions)
Ap132p, by Merck Group (1 mentions)
Western hrp substrate, by Merck Group (1 mentions)
Ab8227, by Abcam (1 mentions)
Ab104274, by Abcam (1 mentions)
Ab125096, by Abcam (1 mentions)
Trypsin inhibitor, by Merck Group (1 mentions)
Trypsin, by Promega (1 mentions)
2 protocols using ab109436
1
Western Blot Analysis of Mitochondrial Proteins
2
Mitochondrial Membrane Proteins Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Isolated mitochondria were incubated with 25 μg/ml trypsin (Promega) on ice in homogenization buffer for the indicated durations. A trypsin inhibitor (Sigma) was added to stop the digestion. Mitochondria were washed with homogenization buffer and subjected to SDS-PAGE. Mitofusin1 and Timm50 were selected as representative outer and inner mitochondrial membrane proteins, respectively. Mitofusin1, Timm50, and mCherry were detected using specific antibodies (ab104274, ab109436, and ab125096, respectively) purchased from Abcam.
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