HRP-conjugated anti-β-ACTIN antibody, Filipin and 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich/ Merck Millipore. Alexa488-conjugated anti-rabbit IgG, HRP-conjugated anti-rabbit total IgG and light chain specific IgG antibodies were purchased from Jackson ImmunoResearch, USA; PE-conjugated F4/80 was procured from Tonbo Biosciences, USA. Alexa Fluor 660 conjugated CD68 was purchased from ThermoFischer Scientific. Anti-G9a, anti-SIRT6, anti-H3K9me1, anti-H3K9me2, anti-H3K9Ac, anti-Ser33/37/Thr41 phospho-β-CATENIN, anti-Ser9 phospho-GSK-3β, anti-β-CATENIN, anti-NRF2, anti-HO1 and anti-TRXR1 antibodies were obtained from Cell Signaling Technology, USA. Anti-LRP2 antibody was purchased from Santa Cruz Biotechnology, USA; anti-SREBP2 antibody was procured from Abcam, USA; and anti-NQO1 antibody was purchased from Calbiochem, USA.
Anti srebp2 antibody
Manufactured by Abcam
Sourced in United States, United Kingdom
The Anti-SREBP2 antibody is a reagent used in research applications. It is designed to detect the SREBP2 protein, which is a transcription factor involved in the regulation of cholesterol and lipid homeostasis. The antibody can be used in various experimental techniques, such as Western blotting, immunohistochemistry, and immunoprecipitation, to study the expression and localization of SREBP2 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti h3k9me2, by Cell Signaling Technology (2 mentions)
Anti sirt6, by Cell Signaling Technology (2 mentions)
Alexa488 conjugated anti rabbit igg, by Jackson ImmunoResearch (2 mentions)
Hrp conjugated anti β actin antibody, by Merck Group (2 mentions)
Anti β catenin, by Cell Signaling Technology (2 mentions)
Anti nrf2, by Cell Signaling Technology (2 mentions)
Filipin, by Merck Group (2 mentions)
Hrp conjugated anti rabbit total igg, by Jackson ImmunoResearch (2 mentions)
Pe conjugated f4 80, by Cytek Biosciences (2 mentions)
Anti h3k9me1, by Cell Signaling Technology (2 mentions)
Anti phospho β catenin ser33 37 thr41, by Cell Signaling Technology (2 mentions)
Anti ser9 phospho gsk 3β, by Cell Signaling Technology (2 mentions)
Anti ho 1, by Cell Signaling Technology (2 mentions)
Anti trxr1, by Cell Signaling Technology (2 mentions)
Anti lrp2 antibody, by Santa Cruz Biotechnology (2 mentions)
Anti g9a, by Cell Signaling Technology (2 mentions)
Anti h3k9ac, by Cell Signaling Technology (2 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Merck Group (2 mentions)
Anti il 1β antibody, by Proteintech (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
7 protocols using anti srebp2 antibody
1
Immunofluorescence Analysis of Cellular Markers
2
SREBP2 and IL-1β Immunohistochemistry
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Tissue samples were prepared and preserved through paraffin embedding and then dewaxed and blocked in a hydrogen peroxide/methanol solution. Antigen retrieval was performed using sodium citrate (pH 6.0) for 2×20 min at 80°C. After that, the sections were incubated with anti-SREBP2 antibody (Abcam, Cambridge, UK) and anti-IL-1β antibody (Proteintech, Chicago, USA) overnight at 4°C, followed by incubation with HRP-conjugated anti-rabbit IgG secondary antibody (Abcam) at 37°C for 30 min. The sections were stained with DAB and counterstained with hematoxylin, dehydrated in ethanol, mounted in dimethylbenzene, and placed under a coverslip. Analysis of IHC images was performed using ImageJ (NIH, Bethesda, USA).
3
Immunofluorescent Imaging of SREBP2
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Cell climbing sheets were gathered, washed with PBS for three times, fixed in 4% paraformaldehyde for 30 min at room temperature, and permeabilized with 0.1% Triton X-100 and blocked with 5% BSA plus Tween 20. Then, the sections were incubated with anti-SREBP2 antibody (diluted at 1:100, Abcam) overnight at 4° C, followed by 1 h of incubation with secondary antibodies at room temperature. Next, the nuclei were counterstained with DAPI (diluted at 1:1,000, Thermo Fisher Scientific) for 10 minutes at room temperature. Finally, fluorescence intensity on the stained sections of aortic roots was measured using ImageJ software.
4
Perforin Expression Regulation by SREBP2
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The antibodies and chemicals used during the current study were as follows: Anti‐SREBP2 antibody (Abcam, Cambridge, UK, ab30682), Perforin (Cell Signaling Technology, Danvers, MA, USA, #3693), GAPDH (Cell Signaling Technology, #5174), Anti‐mouse IgG, HRP‐linked Antibody (Cell Signaling Technology, #7076), Anti‐rabbit IgG, HRP‐linked Antibody (Cell Signaling Technology, #7074), and simvastatin obtained from MCE (HY‐17502). Recombinant human Perforin protein (ab114201) was obtained from Abcam.
5
Western Blot Analysis of Protein Signaling
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Cells were lysed in RIPA buffer, and then lysates were centrifuged at 13,400
g for 10 min and quantified using a BCA protein assay kit (Beyotime, Shanghai, China). Samples were separated on 8%‒12% SDS-PAGE gels and transferred to PVDF membranes (IPVH00010; Millipore, Billerica, USA). The membrane was blocked for 1 h at room temperature with 5% milk solution in TBS-Tween [50 mM Tris (pH 8.0), containing 150 mM NaCl and 0.1% Tween 20] before incubation with primary antibodies, including anti-SREBP2 antibody (Abcam), anti-IL-1β antibody (Proteintech), anti-mTOR antibody, and anti-pmTOR antibody (Cell Signaling Technology, Danvers, USA) at 4°C overnight. Subsequently, HRP-conjugated secondary antibody anti-rabbit IgG was added, followed by incubation at room temperature for 1 h. Western blots were visualized using chemiluminescence reagents (Sigma, St Louis, USA).
g for 10 min and quantified using a BCA protein assay kit (Beyotime, Shanghai, China). Samples were separated on 8%‒12% SDS-PAGE gels and transferred to PVDF membranes (IPVH00010; Millipore, Billerica, USA). The membrane was blocked for 1 h at room temperature with 5% milk solution in TBS-Tween [50 mM Tris (pH 8.0), containing 150 mM NaCl and 0.1% Tween 20] before incubation with primary antibodies, including anti-SREBP2 antibody (Abcam), anti-IL-1β antibody (Proteintech), anti-mTOR antibody, and anti-pmTOR antibody (Cell Signaling Technology, Danvers, USA) at 4°C overnight. Subsequently, HRP-conjugated secondary antibody anti-rabbit IgG was added, followed by incubation at room temperature for 1 h. Western blots were visualized using chemiluminescence reagents (Sigma, St Louis, USA).
6
Immunostaining for Cell Signaling
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HRP-conjugated anti-β-ACTIN antibody, Filipin and 4′,6-Diamidino-2phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich/ Merck Millipore. Alexa488-conjugated anti-rabbit IgG, HRP-conjugated anti-rabbit total IgG and light chain specific IgG antibodies were purchased from Jackson ImmunoResearch, USA; PE-conjugated F4/80 was procured from Tonbo Biosciences, USA. Anti-G9a, anti-SIRT6, anti-H3K9me1, anti-H3K9me2, anti-H3K9Ac, anti-Ser33/37/Thr41 phospho-β-CATENIN, anti-Ser9 phospho-GSK-3β, anti-β-CATENIN, anti-NRF2, anti-HO1 and anti-TRXR1 antibodies were obtained from Cell Signaling Technology, USA. Anti-LRP2 antibody was purchased from Santa Cruz Biotechnology, USA; anti-SREBP2 antibody was procured from Abcam, USA; and anti-NQO1 antibody was purchased from Calbiochem, USA.
7
Lipid Metabolism Protein Analysis
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Nuclear extracts were prepared using a previously reported method 34) . Western blot analysis was performed as previously described 35) . The following antibodies were used: anti-UFM1 antibody, anti-SREBP2 antibody, anti-SR-A antibody, anti-CD36 antibody, anti-SR-BI antibody, anti-ABCA1 antibody, and anti-ABCG1 antibody were purchased from Abcam (1:1000; Cambridge, MA, USA); anti-LXRα antibody, anti-LDL antibody, and anti-HMGCR antibody were obtained from Santa Cr uz Biotechnology Inc. (1:1000; Santa Cruz, CA,USA); anti-β-actin antibody, anti-αtubulin antibody, and anti-histone H1 antibody (1:2000; Sigma) were used as the loading control.
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