Ab13847

Manufactured by Merck Group

Ab13847 is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use. The core function of this product is to facilitate standard laboratory procedures. No further details are available without making interpretations or extrapolations.

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Lab products found in correlation

Gapdh, by Merck Group (1 mentions) Ab38238, by Abcam (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Epac2, by Cell Signaling Technology (1 mentions) Rpn303f, by GE Healthcare (1 mentions) Goat anti mouse fab fragment, by Jackson ImmunoResearch (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Triton x 100, by Avantor (1 mentions) Prb 145p, by Fortrea (1 mentions)

2 protocols using ab13847

Western Blot Analysis of Protein Expression

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The cortical regions of the brains were collected and homogenized in the lysis buffer containing protease inhibitor. The proteins were extracted and the protein concentration was measured using a BCA protein assay kit (Thermo, 23227). Equal quantities of protein (40 μg) from each samples were loaded for SDS-PAGE. After electrophoresis, the protein was transferred onto polyvinylidene difluoride membranes (GE Healthcare, RPN303F). The membranes were blocked with 5% nonfat milk for 1 h at room temperature and subsequently incubated overnight at 4°C with the following primary antibodies: Epac2 (1:1,000, Cell Signaling, 43239), P38 (1:1,000, Abcam, ab17009), P-P38 (1:500, Abcam, ab38238), caspase-3 (1:1,000, Abcam, ab13847), GAPDH (1:10,000, Sigma, G9545). The membranes were incubated with appropriate secondary antibodies for 2 h at room temperature. The target band signals were detected by Enhanced chemiluminescence (ECL) method. The signals were quantified using Image J software.

Zhang L., Zhang L., Liu H., Jiang F., Wang H., Li D, & Gao R. (2018). Inhibition of Epac2 Attenuates Neural Cell Apoptosis and Improves Neurological Deficits in a Rat Model of Traumatic Brain Injury. Frontiers in Neuroscience, 12, 263.

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Immunohistochemical Staining of Skin Tissue

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Similar to hematoxylin and eosin staining, we performed a series of tissue incubations in xylene and ethanol to remove paraffin and hydrate the tissue. Following this step, we performed antigen retrieval in sodium citrate (pH6.0) and permeabilization in Triton X-100 (VWR). We than performed a series of washing steps to remove the detergent. We then performed two blocking steps, each lasting one hour. The first blocking step was 10% BSA in PBS. After two brief washing steps in PBS, we than incubated the slides in 40 μg/ml of goat anti-mouse Fab fragment in PBS (Jackson ImmunoResearch Laboratories, 115-007-003). Primary antibodies were incubated for 18–24 hours at 4°C. Primary antibodies included TP63 (Santa Cruz, 4A4, sc-8431), Keratin 6 (Covance, PRB-169P), Keratin 14 (Novocastra, NCL-L-LL002), IRF6 (Sigma-Aldrich, SAB2102995), Keratin 1 (NCL-CK1, Novocastra), Loricrin (PRB-145P, Covance), Ki-67 (ab15580, Abcam), GRHL3 (a kind gift from Dr. B. Andersen, UCI, Irvine, CA), Activated Caspase 3 (Abcam, Ab13847), KRT17 (Sigma-Aldrich, HPA000453) JAG2 (ThermoFisher, PI701287) and MMP13 (Sigma-Aldrich, HPA030636). Secondary antibodies (goat anti-mouse and goat anti-rabbit (Molecular Probes)) were incubated for 1–2 hours. Nuclei were labeled with DAPI (Invitrogen).

Kousa Y.A., Moussa D, & Schutte B.C. (2017). IRF6 expression in basal epithelium partially rescues Irf6 knockout mice. Developmental dynamics : an official publication of the American Association of Anatomists, 246(9), 670-681.

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