A27036

Manufactured by Thermo Fisher Scientific

The A27036 is a laboratory equipment product from Thermo Fisher Scientific. It is designed for general laboratory use. Further details on the core function and specifications of this product are not available.

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Lab products found in correlation

Calnexin, by Santa Cruz Biotechnology (1 mentions) Gm130, by Cell Signaling Technology (1 mentions) Mini protean tgx precast gel, by Bio-Rad (1 mentions) Supersignal west femto substrate, by Thermo Fisher Scientific (1 mentions) T6074, by Merck Group (1 mentions) Wet method, by Bio-Rad (1 mentions) Sc 13029, by Santa Cruz Biotechnology (1 mentions) Enhanced chemiluminescence ecl reagent, by Thermo Fisher Scientific (1 mentions) Bca kit, by Thermo Fisher Scientific (1 mentions) Pierce ip lysis buffer, by Thermo Fisher Scientific (1 mentions) Ab32503, by Abcam (1 mentions) Ab209847, by Abcam (1 mentions) Ab92552, by Abcam (1 mentions) Ab32124, by Abcam (1 mentions)

4 protocols using a27036

Immunofluorescence and Western Blot Protocols for JCPyV VP1 Antibody Characterization

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PAB597 is a hybridoma supernatant that produces a monoclonal antibody against JCPyV VP1 [61 (link)]. AF488 labeled goat anti-mouse 488 or anti-rabbit antibodies (Thermo/Invitrogen) were used as the secondary antibodies for immunofluorescence experiments. Transthyretin primary antibody (Dako, A0002) was used for initial characterization of CPE cells (1:250, overnight at 4C) followed by AF488 labeled goat anti-rabbit antibody (1:1,000). Primary antibodies and the respective dilutions used for Western blotting included annexin V (Abcam, ab117439, 1:1,000), CD9 (Cell Signaling Technologies, CST 13174S, 1:1,000), flotillin-1 (CST 18634S, 1:1,000), GM130 (Cell Signaling Technologies, CST 12480, 1:1,000), calnexin (Santa Cruz Biotechnology, TX; sc-11397, 1:200), cytochrome c (BD Pharmingen 556433, 1:500), TSG101 (Thermo PA5-31260, 1:500), and PAB597 (1:2,000). Secondary antibodies used for Western blotting anti-mouse horseradish peroxidase (HRP) (Thermo A28177) and anti-rabbit HRP (Thermo A27036), both used at 1:10,000.

O’Hara B.A., Morris-Love J., Gee G.V., Haley S.A, & Atwood W.J. (2020). JC Virus infected choroid plexus epithelial cells produce extracellular vesicles that infect glial cells independently of the virus attachment receptor. PLoS Pathogens, 16(3), e1008371.

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Dual Antibody Immunoblotting Procedure

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A27036, Thermo Fisher, dilution 1 : 2000 (WB), superclonal, goat/IgG, anti‐rabbit, HRP NXA931, Cytiva, dilution 1 : 2000 (WB), polyclonal, sheep/IgG, anti‐mouse, HRP.

Jänsch M., Lubomirov L.T., Trum M., Williams T., Schmitt J., Schuh K., Qadri F., Maier L.S., Bader M, & Ritter O. (2022). Inducible over‐expression of cardiac Nos1ap causes short QT syndrome in transgenic mice. FEBS Open Bio, 13(1), 118-132.

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Western Blot Analysis of Protein Expression

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Cells were lysed directly in 1× Laemmli buffer, and proteins were separated through denaturing SDS-PAGE electrophoresis using Mini-Protean TGX precast gels (Bio-Rad) and transferred to PVDF membrane using the wet method (Bio-Rad). Membranes were blocked with 5% skim milk in TBS-1% Tween (TBST) for 1 h at RT and probed overnight at 4°C with primary antibodies in TBST and 5% BSA. The following antibodies were used: mouse anti-Srf (1/1,000; sc 13029; Santa Cruz Biotechnology), rabbit anti–pan actin (1/750; AAN01-A; Cytoskeleton), and mouse anti–α tubulin (1/4,000; T 6074; Sigma-Aldrich). After washing in TBST, membranes were hybridized with goat anti-mouse and goat anti-rabbit secondary antibodies coupled to HRP (1/10,000; 62-6520 and A27036; Thermo Fisher Scientific). Proteins were revealed using SuperSignal West Femto substrate (Thermo Fisher Scientific).

Randrianarison-Huetz V., Papaefthymiou A., Herledan G., Noviello C., Faradova U., Collard L., Pincini A., Schol E., Decaux J.F., Maire P., Vassilopoulos S, & Sotiropoulos A. (2018). Srf controls satellite cell fusion through the maintenance of actin architecture. The Journal of Cell Biology, 217(2), 685-700.

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Western Blot Analysis of Cellular Proteins

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After lysis using Pierce IP lysis buffer (Thermo Fisher), total protein concentration was measured with a BCA kit (Thermo Fisher). Then, the protein samples were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to PVDF membranes. After sealing in 5% skim milk, probing with the primary antibodies against PCNA (ab92552, 1:1000, Abcam), Bcl-2 (ab32124, 1:1000, Abcam), Bax (ab32503, 1:1000, Abcam), HK2 (ab209847, 1:1000, Abcam), FOXN3 (#711585, 1:100, Thermo Fisher), GAPDH (#5174, 1:1000, CST, USA) at 4°C overnight. Subsequently, a secondary antibody (#A27036, 1:4000, Thermo Fisher) was applied. The bands were acquired using the enhanced chemiluminescence (ECL) reagent (Thermo Fisher).

Chen X., Dou B., An J., Feng S., Liu N., & Yao S. (2021). MAGI2-AS3 modulates proliferation, apoptosis and glycolysis in acute lymphoblastic leukemia via miR-452-5p/FOXN3 axis.

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