Odyssey blocker pbs

Manufactured by LI COR

The Odyssey Blocker PBS is a ready-to-use blocking buffer designed for use with the Odyssey Infrared Imaging System. It is formulated to minimize non-specific binding in Western blot and other immunoassay applications. The Odyssey Blocker PBS provides consistent and reliable results.

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Lab products found in correlation

Irdye 800cw goat anti rabbit, by LI COR (2 mentions) Anti lamin a c e1, by Santa Cruz Biotechnology (2 mentions) Halt phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Nupage 3 8 tris acetate gel, by Thermo Fisher Scientific (1 mentions) Odyssey imager, by LI COR (1 mentions) Iblot apparatus, by Thermo Fisher Scientific (1 mentions) Halt protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions) Odyssey clx imaging system, by LI COR (1 mentions) Image studio software version 5, by LI COR (1 mentions) Rabbit anti ha c29f4, by Cell Signaling Technology (1 mentions) Bradford protein assay, by Bio-Rad (1 mentions) Irdye 680lt goat anti mouse, by LI COR (1 mentions) Anti arid1a psg3, by Santa Cruz Biotechnology (1 mentions) Anti arid2 d8d8u, by Cell Signaling Technology (1 mentions) Odyssey clx, by LI COR (1 mentions) Anti pias1 d33a7, by Cell Signaling Technology (1 mentions) Anti ha c29f4, by Cell Signaling Technology (1 mentions)

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Odyssey blocker (3 citations)

4 protocols using odyssey blocker pbs

Western Blot Analysis of FANCD2

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HT1080 cells treated as indicated were washed twice with ice-cold PBS, collected in a centrifugation tube, and lysed on ice for 0.5 h with RIPA buffer supplemented with Halt Protease Inhibitor Cocktail and Halt Phosphatase Inhibitor Cocktail (Thermo Fisher Scientific, Waltham, MA). The protein concentration was determined by performing a BCA assay (Thermo Fisher Scientific) according to the manufacturer’s recommendation. Normalized amounts of samples were separated in NuPAGE 3–8% Tris-Acetate Gel (Thermo Fisher Scientific) and electrotransferred to a nitrocellulose membrane by using an iBlot apparatus (Thermo Fisher Scientific). The membrane was blocked with Odyssey Blocker (PBS) (Li-Cor, Lincoln, NE) and incubated with anti-FANCD2 antibody (Fl-17, sc-20022, Santa Cruz Biotechnology, Dallas, TX) at a dilution of 1:200 in the same blocking buffer at 4°C overnight, rinsed with TBS-0.05% (v/v) Tween 20 three times for 10 min each, incubated with goat anti-rabbit IRDye 800CW (Li-Cor, 10:000) at room temperature for 1 h, and then rinsed with TBS-0.05% Tween 20 three times for 10 min each. Protein bands were detected by fluorescence using an Odyssey Imager (Li-Cor).

Fujii N., Evison B.J., Actis M.L, & Inoue A. (2015). A novel assay revealed that ribonucleotide reductase is functionally important for interstrand DNA crosslink repair. Bioorganic & medicinal chemistry, 23(21), 6912-6921.

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Western Blotting Immunodetection Procedure

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In all, 15 µg of protein was loaded into each well of an SDS-PAGE gel and transferred onto a PVDF membrane. Blots were blocked at room temperature for 1 hour in Odyssey Blocker PBS (Li-COR) + 2.5% BSA. Blots were incubated with primary antibody overnight at 4 °C and with secondary antibody (goat anti-mouse, Li-COR, 926-68020, 1:20,000 and/or goat anti-rabbit, Li-COR, 926-32211, 1:20,000) for 1 hour at room temperature. Li-COR Odyssey Clx imaging system was used to image blots. Image Studio Software version 5.2 (Li-COR) was used for blot analysis. Antibodies used for western blotting included mouse monoclonal anti-Lamin A/C (E1) (Santa Cruz Biotechnology, sc-376248, 1:1000), rabbit polyclonal anti-AFF1 (Bethyl, A-3020344A-T, 1:500), rabbit polyclonal anti-AFF4 (Abclonal, A4644, 1:1000), rabbit polyclonal anti-HEXIM1 (Bethyl, A303-112A-T, 1:1000), rabbit monoclonal anti-ATF-3 (E9J4N) (Cell Signaling, 18665, 1:1000), rabbit monoclonal anti-DUSP1 (E8L7D) (Cell Signaling, 48625, 1:1000), mouse monoclonal anti-RhoE (RND3) (Cell Signaling, 3664, 1:300), Rabbit anti-PhosphoThr186 CDK9 (Cell Signaling, 2549, 1:1000), Rabbit anti-CDK9 (C12F7) (Cell Signaling, 2316, 1:1000), Rabbit anti-HA (C29F4) (Cell Signaling, 3724, 1:1000), Rabbit anti-DNMT1 (D63A6) (Cell Signaling, 5032, 1:1000), and Rabbit anti-MYC (D84C12) (Cell Signaling, 5605, 1:1000).

Lloyd S.M., Leon D.B., Brady M.O., Rodriguez D., McReynolds M.P., Kweon J., Neely A.E., Blumensaadt L.A., Ho P.J, & Bao X. (2022). CDK9 activity switch associated with AFF1 and HEXIM1 controls differentiation initiation from epidermal progenitors. Nature Communications, 13, 4408.

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Quantification of Nuclear Pore Proteins

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The protein concentration of keratinocyte lysate was measured by BioRad Bradford Protein Assay, and 10–15 ug µg of protein per sample was loaded for SDS-PAGE. The separated proteins were transferred onto a (polyvinylidene difluoride) PVDF membrane. After blocking using the Odyssey Blocker PBS (LI-COR) + 2.5% BSA for 1 h at the room temperature, primary antibodies were added to the PVDF membrane for overnight incubation at 4 °C. The primary antibodies used in this study include NUP205 (H1) (Santa Cruz sc-377047, 1:200), NUP93 (E-8) (Santa Cruz sc-374399, 1:200), NUP133 (Santa Cruz sc-376699, 1:200), lamin A/C (ThermoFisher MA5-35284, 1:1000; or Santa Cruz sc-376248, 1:1000). Secondary antibody (Goat anti-mouse LI-COR 96-68020, Goat anti-mouse LI-COR 96-32211) incubation was performed at room temperature for 1 h at 1:15000 dilution.

Neely A.E., Zhang Y., Blumensaadt L.A., Mao H., Brenner B., Sun C., Zhang H.F, & Bao X. (2023). Nucleoporin downregulation modulates progenitor differentiation independent of nuclear pore numbers. Communications Biology, 6, 1033.

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Immunoblot Analysis of Chromatin Remodeling Proteins

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For immunoblot analysis, 10–30 μg of cell lysate was loaded per lane for SDS-page and then transferred to PVDF membranes. Blots were blocked with Odyssey Blocker PBS (Li-COR) + 2.5% BSA at room temperature for 1 h. Blots were then incubated with primary antibody at 4°C overnight and then with secondary antibodies (IRDye 680LT goat anti-mouse, LI-COR, 926–68020, 1:15,000, and/or IRDye 800CW goat anti-rabbit, LI-COR, 926–32211, 1:15,000) at room temperature for 1 h. The blots were imaged using Li-COR Odyssey CLx (LI-COR). Primary antibodies used for western blotting include anti-PBRM1 (Bethyl, A700-019), anti-PIAS1 (D33A7) (Cell Signaling, 3550), anti-ARID2 (D8D8U) (Cell Signaling, 82342), anti-BRG1 (G-7) (Santa Cruz Biotechnology, sc-17796), anti-ARID1A (PSG3) (Santa Cruz Biotechnology, sc-32761), anti-BRD7 (Bethyl, A302-304A-T), anti-HA (C29F4) (Cell Signaling, 3724), and anti-Lamin A/C (E−1) (Santa Cruz Biotechnology, sc-376248).

Ho P.J., Kweon J., Blumensaadt L.A., Neely A.E., Kalika E., Leon D.B., Oh S., Stringer C.W., Lloyd S.M., Ren Z, & Bao X. (2024). Multi-omics integration identifies cell-state-specific repression by PBRM1-PIAS1 cooperation. Cell Genomics, 4(1), 100471.

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