Total protein was extracted using cell lysis buffer (20 mM Tris, pH 7.5, 150 mM NaCl, 1 % Triton X-100, 0.1 mM PMSF) and quantified using a BCA Protein Assay Reagent Kit (Pierce, Rockford, IL, USA). Then, 20 µg total protein was subjected to 12 % sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinyl difluoride (PVDF) membrane, which was and blocked with 5 % skim milk in TBST buffer at 37 °C for 1 h. Membranes were incubated overnight with gentle shaking at 4 °C with rabbit anti-PRRSV N protein antibody (diluted at 1:2500, donated by Dr. Yuanpeng Zhang) and mouse anti-β-actin antibody (1:200, Boster, China). After washing, membranes were incubated with an appropriate secondary antibody (1:10,000, SunShineBio, China) for 1 h at 37 °C. Antibodies were visualized using enhanced chemiluminescence (ECL) reagent (Boster, China) and images were recorded using an ImageQuant LAS 4000 (GE Healthcare, Sweden).
Mouse anti β actin antibody
Manufactured by Boster Bio
Sourced in China
The Mouse anti-β-actin antibody is a primary antibody used for the detection and quantification of the β-actin protein in various biological samples. β-actin is a highly conserved cytoskeletal protein that is ubiquitously expressed in eukaryotic cells and is often used as a loading control or reference protein in Western blot analysis and other immunoassays.
Automatically generated - may contain errors
Lab products found in correlation
Imagequant las 4000, by GE Healthcare (2 mentions)
Ecl reagent, by Boster Bio (1 mentions)
Radioimmunoprecipitation assay lysis buffer, by CWBIO (1 mentions)
Rabbit anti cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti pk2 antibody, by Abcam (1 mentions)
Hrp conjugated goat anti mouse igg, by Boster Bio (1 mentions)
Urine protein assay kit, by Nanjing Jiancheng (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Boster Bio (1 mentions)
Goat anti mouse secondary antibody, by Boster Bio (1 mentions)
Anti desmin, by Abcam (1 mentions)
Anti nephrin, by Abcam (1 mentions)
5 protocols using mouse anti β actin antibody
1
Western Blot Analysis of PRRSV N Protein
2
Western Blot Antibody Panel for Cell Signaling Proteins
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Rabbit anti-TERT polyclonal antibody (#abs136649; 1:1500) was obtained from Absin. Rabbit anti-ETS1 polyclonal antibody (#A00931; 1:1500); mouse anti-β-Actin antibody (BM5422; 1:1500); mouse anti-β-tubulin antibody (#BM3877; 1:1500); rabbit polyclonal anti-PTEN antibody (#PB0423; 1:1500); rabbit polyclonal anti-Catenin-β antibody (#BM3905; 1:1500); mouse anti-c-MYC antibody (#BM0238; 1:1500); rabbit anti-BLC2 monoclonal antibody (#BM4985; 1:1500); rabbit anti-BAX monoclonal antibody (#BM3964; 1:1500); and rabbit polyclonal anti-VDAC1 antibody (#BA3754; 1:1500) were obtained from Boster Biological Technology. Rabbit anti-GABPA polyclonal antibody (#21542-1-AP; 1:1500); rabbit anti-GABPB1 polyclonal antibody (#12597-1-AP; 1:1500); rabbit caspase 9 polyclonal antibody (#10380-1-AP; 1:1500); rabbit ANT1/2 polyclonal antibody (#17796-1-AP; 1:1500); and rabbit cytochrome C polyclonal antibody (#10993-1-AP; 1:1500) were obtained from Proteintech. AIF rabbit monoclonal antibody (#AF1273; 1:1500) was obtained from Beyotime.
3
Testicular Protein Expression Analysis
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The testicular tissue was lysed on ice with radioimmunoprecipitation assay lysis buffer (Cwbio, Taizhou, China). Subsequently, 20–40 μg of protein was electrophoresed on an 8%–12% (w/v) sodium dodecyl sulfate-polyacrylamide gel gradient and transferred to a nitrocellulose membrane. The membranes were blocked with 5% (w/v) nonfat milk for 1 h in Tris-buffered saline (TBS) and subsequently incubated with the following primary antibodies: rabbit anti-PK2 antibody (1:200, Abcam, Cambridge, MA, USA), rabbit anti-cleaved-caspase-3 antibody (1:1000, Cell Signaling Technology [CST], Boston, MA, USA), rabbit anti-caspase-8 antibody (1:1000, CST), rabbit anti-Bax antibody (1:1000, CST), rabbit anti-Bcl-2 antibody (1:1000, CST), and mouse anti-β-actin antibody (1:500, Boster, Wuhan, China). Immunoreactive bands were detected using electrochemiluminescence (ECL; Pierce, Waltham, MA, USA).
4
PRRSV GP5 Protein Detection by Western Blot
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Cells were lysed in cell lysis buffer (20 mM Tris, pH 7.5, 150 mM NaCl, 1% Triton X-100, 0.1 mM phenylmethylsulfonyl fluoride). Protein content was measured using the BCA Protein Assay Reagent Kit (Pierce, Rockford, IL, USA) and 20 µg total protein was loaded on a 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE) gel, transferred to a polyvinyl difluoride (PVDF) membrane and blocked with TBS containing 5% skim milk and 0.1% Tween-20 at 37°C for 2 h. Membranes were incubated overnight with gentle shaking at 4°C with mouse anti-PRRSV GP5 antibody (1:500, TONGDIAN, Hangzhou, China) or mouse anti-β-actin antibody (1:200, Boster, Wuhan, China). After washing, membranes were incubated with HRP-conjugated goat anti-mouse IgG (1:5,000, Boster, Wuhan, China) for 1 h at 37°C. Antibodies were visualized using an enhanced chemiluminescence reagent and images were recorded using a ImageQuant LAS 4000 instrument (GE Healthcare, Sweden).
5
Immunohistochemical Analysis of Kidney Injury
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Acupuncture needles were purchased from Suzhou Medical Sino-Foreign Joint Venture Suzhou Hua Tuo Medical Instruments Co., Ltd. STZ was purchased from Sigma-Aldrich Chemical Co. (MO, USA). Urine protein assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Anti-nephrin, anti-podocalyxin, and anti-desmin antibodies were purchased from Abcam Biotechnology (CA, USA). Anti-CD2AP antibody was purchased from R&D Biotechnology (MN, USA), and mouse anti-β-actin antibody was purchased from Boster Biotechnology (Wuhan, China). Horseradish peroxidase-conjugated goat anti-rabbit IgG and goat anti-mouse secondary antibodies were purchased from Boster Biotechnology (Wuhan, China). Rabbit anti-sheep IgG was purchased from EarthOx Life Sciences (CA, USA). Finally, a chemiluminescence kit was purchased from CWBIO (Beijing, China)
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