Cell lysates were incubated with primary antibodies against BVRA (sc-393385; Santa Cruz Biotechnology, Inc., CA, USA), ERK2 (ab32081; Abcam, Shanghai, China) or normal mouse IgG (sc-2025; Santa Cruz Biotechnology, Inc., CA, USA) at 4°C overnight, followed by incubating with protein A/G Plus-Agarose beads (20423; Thermo Fisher Scientific, MA, USA) at 4°C for 2 h. The beads were then boiled for western blot analysis.
Sc 393385
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Sc-393385 is a laboratory product offered by Santa Cruz Biotechnology. It is a piece of equipment designed for use in scientific research and experiments. The core function of this product is to facilitate specific laboratory processes, though a detailed description cannot be provided while maintaining an unbiased and strictly factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab150113, by Abcam (3 mentions)
Ab32081, by Abcam (2 mentions)
Ab1791, by Abcam (2 mentions)
Ab92946, by Abcam (2 mentions)
Ab8245, by Abcam (2 mentions)
Ab150077, by Abcam (2 mentions)
Protein a g plus agarose beads, by Thermo Fisher Scientific (1 mentions)
Sc 2025, by Santa Cruz Biotechnology (1 mentions)
Ab109199, by Abcam (1 mentions)
Ab199728, by Abcam (1 mentions)
Ab68477, by Abcam (1 mentions)
Ab16663, by Abcam (1 mentions)
Ab184699, by Abcam (1 mentions)
Ab201015, by Abcam (1 mentions)
Ab211542, by Abcam (1 mentions)
Immunofluorescence microscope, by Zeiss (1 mentions)
Beyoecl plus kit, by Beyotime (1 mentions)
Ab182858, by Abcam (1 mentions)
Ab32503, by Abcam (1 mentions)
Ab32518, by Abcam (1 mentions)
4 protocols using sc 393385
1
Immunoprecipitation and Western Blot
2
Western Blot Analysis of Cell Signaling Proteins
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Protein samples from LECs of each group were added to SDS-PAGE and blotted onto PVDF membrane. Primary antibodies against p21 (ab109199; Abcam, 1 : 1000), CDKN2A/p16INK4α (ab211542; Abcam, 1 : 1000), Cyclin D1 (ab16663; Abcam, 1 : 1000), CDK4 (ab199728; Abcam, 1 : 1000), phosphorylated-ERK1/2 (ab201015; Abcam, 1 : 1000), ERK1/2 (ab184699; Abcam, 1 : 1000), BVRA (sc-393385; Santa Cruz, 1 : 1000), ERK2 (ab32081; Abcam, 1 : 1000), Nrf2 (ab92946; Abcam, 1 : 1000), HO-1 (ab68477; Abcam, 1 : 1000), GAPDH (ab8245; Abcam, 1 : 5000), and Histone H3 (ab1791; Abcam, 1 : 5000) were incubated with blots at 4°C overnight, followed by incubation with goat anti-mouse IgG H&L (ab150113; Abcam, 1 : 5000) or goat anti-rabbit IgG H&L (ab150077; Abcam, 1 : 5000) for 1 h at room temperature. GAPDH and Histone H3 were used as the quantitative loading control.
3
Hydrogen Peroxide-Induced LEC Oxidative Stress
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5 × 104/well LECs were seeded in 24-well plates and treated with 200 μM H2O2 for 4 h. Then, samples were fixated and incubated with primary antibody against BVRA (sc-393385; Santa Cruz, 1 : 200) and goat anti-mouse IgG H&L (ab150113; Abcam, 1 : 5000). The reaction was detected using immunofluorescence microscope (Zeiss, German).
4
Evaluation of NF-κB and Antioxidant Pathways
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LECs were lysed, and protein concentration was evaluated using BCA protein assay kit (P0012, Beyotime Biotechnology, Shanghai, China). Then, protein samples were added to SDS-PAGE and blotted onto PVDF membrane. Primary antibodies against NF-κB p65 (ab16502; Abcam, 1:1000), IκB (ab32518; Abcam, 1:1000), iNOS (ab178945; Abcam, 1:1000), Bax (ab32503; Abcam, 1:1000), Bcl-2 (ab182858; Abcam, 1:1000), Caspase-3 (ab184787; Abcam, 1:1000), BVRA (sc-393385; Santa Cruz, 1:1000), Nrf2 (ab92946; Abcam, 1:1000), HO-1 (ab52947; Abcam, 1:1000), GAPDH (ab8245; Abcam, 1:5000), and histone H3 (ab1791; Abcam, 1:5000) were incubated with blots at 4°C overnight. Then, blots were incubated with goat anti-mouse IgG H&L (ab150113; Abcam, 1:5000) or goat anti-rabbit IgG H&L (ab150077; Abcam, 1:5000) for 1 h at room temperature. The blots were detected by BeyoECL Plus kit (P0018S, Beyotime Biotechnology, Shanghai, China) and analyzed using Image J software. GAPDH was used as a cytosolic marker and H3 a nuclear marker.
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