A01020

Manufactured by Abbkine

Sourced in United States

A01020 is a laboratory equipment product. It is designed for use in scientific research and experimentation. The core function of this product is to [description not available].

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Lab products found in correlation

Gapdh, by Abbkine (3 mentions) Peroxidase conjugated goat anti mice igg, by Jackson ImmunoResearch (2 mentions) Peroxidase conjugated goat anti rabbit igg, by Jackson ImmunoResearch (2 mentions) A21010, by Abbkine (2 mentions) Ab203043, by Abcam (1 mentions) Ab93048, by Abcam (1 mentions) Labimage software, by Bio-Rad (1 mentions) A01030hrp, by Abbkine (1 mentions) Ab109226, by Abcam (1 mentions) A01010, by Abbkine (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) A21020, by Abbkine (1 mentions) Bca assay kit, by Applygen (1 mentions) Sc 515777, by Santa Cruz Biotechnology (1 mentions) Quantitative protein kit, by Applygen (1 mentions) Enhanced chemoluminescence, by Beyotime (1 mentions) Kisspeptin, by Abcam (1 mentions) β tubulin, by Abbkine (1 mentions) A01030, by Abbkine (1 mentions)

7 protocols using a01020

Protein Expression Analysis of Intestinal and Hypothalamic Tissues

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The total protein from hypothalamus and colon were separated by SDS-PAGE and then transferred to PVDF membrane. The membrane was soaked with skimmed milk powder to reduce the binding of non-specific antibodies. The membranes were then exposed to primary antibodies and secondary antibodies (peroxidase-conjugated goat anti-rabbit IgG (111–035-003, Jackson) or peroxidase-conjugated goat anti-mice IgG (115–035-003, Jackson) depending on the primary antibody). Protein was quantified with Lab image Software (BioRad, USA) and expressed as relative units to housekeeping proteins.
The primary antibodies are as follows: anti-free fatty acid receptor 2 (FFAR2; ABC299, Merck Millipore), anti-monocarboxylic acid transporter 1 (MCT1, ab93048, Abcam), anti-small peptide transporters (PEPT1, ab203043, Abcam), anti-GAPDH (A01020, Abbkine), and anti-β-tubulin (A01030HRP, Abbkine).

Bo T., Liu H., Liu M., Liu Q., Li Q., Cong Y., Luo Y., Wang Y., Yu B., Pu T., Wang L., Wang Z, & Wang D. (2023). Mechanism of inulin in colic and gut microbiota of captive Asian elephant. Microbiome, 11, 148.

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Intestinal Protein Expression Analysis

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The intestinal tissues were homogenized in a RIPA extraction buffer with a protease inhibitor cocktail (Roche) for 30 min. Proteins were quantified via a BCA protein assay kit (Beyotime Biotechnology, China), and then prepared in Laemmli buffer and boiled in water bath. Equal amounts of tissue lysates (50 mg) were loaded and separated on10% SDS-PAGE gels with constant voltage of 100 V for 100 min, and transferred to PVDF membranes with constant current of 300 mA for 80 min in ice bath. Membranes were subsequently blocked in 8% (w/v) non-fat dried milk for 1 h at room temperature, then incubated overnight at 4 °C with specific antibodies against tryptase (Abcam, ab109226, 1:2000), PAR2 (Abcam, ab154835, 1:1000), β-actin(Abbkine, A01010, 1:2000) and GAPDH (Abbkine, A01020, 1:2000). After fully washing, the membranes were probed with HRP-conjugated goat anti-rabbit or mouse antibody (1:5000, Pierce, USA) for 1 h at room temperature. Then, the protein bands were developed in the SuperSignal West Pico Substrate (Pierce, USA) and quantified by the FluorChem FC3 software (ProteinSimple, USA).

Zhang L., Song J., Bai T., Qian W, & Hou X.H. (2017). Stress induces more serious barrier dysfunction in follicle-associated epithelium than villus epithelium involving mast cells and protease-activated receptor-2. Scientific Reports, 7, 4950.

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Protein Extraction and Western Blot Analysis

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Protein extraction was carried out. Briefly, a 1-mm segment was cut from ISR tissue. Loose connective tissue was removed, 200 μL of ice-cold radio immunoprecipitation assay (RIPA) lysis buffer and 2 μL of phenylmethylsulfonyl fluoride were added, and the tissue segment was homogenized. The homogenate was centrifuged at 12 000 g for 30 minutes at 4°C, and the supernatant was collected. The protein concentration was determined was determined using a bicinchoninic acid (BCA) assay kit (p1511; Applygen, Beijing, China) following manufacturer instructions. Equal amounts of protein were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene fluoride (PVDF) membranes. The latter were washed with TBST (tris-buffered saline and Tween 20), blocked with 10% skimmed milk for 2 hours, and incubated with the same primary antibodies (1:1000 dilution) employed for immunohistochemical staining. PVDF membranes were washed in TBST buffer and probed with secondary antibodies: goat anti-rabbit immunoglobulin (Ig)G (1:4000; A21020; Abbkine Scientific, Beijing, China) and goat anti-mouse IgG (1:4000; A21010; Abbkine Scientific) and visualized using an enhanced chemiluminescence (ECL) kit (Beyotime, Shanghai, China). Glyceraldehyde 3-phosphate dehydrogenase (GAPHD; A01020; Abbkine Scientific) was used as an endogenous control.

Song J.B., Shen J., Fan J., Zhang Z., Yi Z.J., Bai S., Mu X.L, & Xiao L. (2020). Effects of a Matrix Metalloproteinase Inhibitor-Eluting Stent on In-Stent Restenosis. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 26, e922556-1-e922556-17.

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Western Blot Analysis of cGAS and GAPDH

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Western blotting analysis was conducted as previously described [65 (link)]. Primary antibodies against cGAS (Mouse, 1:1000, sc-515777, Santa Cruz Biotechnology) and, GAPDH (GAPDH, A01020, Abbkine, Redlands, CA, USA) were used. ImageJ software (NIH) was used to analyze protein expression levels which were normalized to GAPDH.

Lu G.F., Chen S.C., Xia Y.P., Ye Z.M., Cao F, & Hu B. (2021). Synergistic inflammatory signaling by cGAS may be involved in the development of atherosclerosis. Aging (Albany NY), 13(4), 5650-5673.

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Protein Extraction and Western Blotting

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Briefly, proteins were extracted from specimens with the stents removed. Protein concentration was measured with a quantitative protein kit (Applygen, Beijing, China). An equivalent of 30 mg total protein was used for Western blotting analysis based on standard procedures. Primary antibodies, from the same source as for the immunohistochemical analysis, MMP-2 and MMP-9 were used. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Mouse IgG1 GAPDH Monoclonal antibody, A01020 [Abbkine, Watlham, MA, USA]) protein was used as a loading control.

Shen J., Song J.B., Fan J., Zhang Z., Yi Z.J., Bai S., Mu X.L., Yang Y.B, & Xiao L. (2021). Distribution and Dynamic Changes in Matrix Metalloproteinase (MMP)-2, MMP-9, and Collagen in an In Stent Restenosis Process. European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery, 61(4).

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Western Blot Analysis of Hypothalamic Proteins

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Hypothalamus was homogenized in Radio Immunoprecipitation Assay Lysis buffer (RIPA) buffer and cleared by centrifugation, according to the standard techniques. Western blots of whole tissue lysates were probed with primary antibodies against Kisspeptin (ab226786, Abcam, Cambridge, United Kingdom), GnRH (PA5-97047, Thermo Fisher Scientific, Waltham, MA, United States), β-Tubulin (A01030, Abbkine, CA, United States), GAPDH (A01020; Abbkine, CA, United States). The secondary antibody used was either peroxidase-conjugated goat anti-rabbit IgG (111-035-003; Jackson, West Grove, PA, United States), or peroxidase-conjugated goat anti-mice IgG (115-035-003; Jackson, West Grove, PA, United States). Protein markers (20351ES76; Shanghai Yisheng, China) were added on both sides of each gel to verify bands. The polyvinylidene fluoride (PVDF) membranes were detected by enhanced chemoluminescence (Beyotime, China). The bands were analyzed using Image LabTM Software (Bio-Rab Laboratories, Hercules, CA, United States), were normalized to β-Tubulin or GAPDH, and were expressed as relative units (RU).

Bo T., Liu M., Tang L., Lv J., Wen J, & Wang D. (2022). Effects of High-Fat Diet During Childhood on Precocious Puberty and Gut Microbiota in Mice. Frontiers in Microbiology, 13, 930747.

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Western Blot Analysis of HDAC3 Expression

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Protein samples were prepared in RIPA buffer on ice. Lysates were separated by centrifugation at 10,000×g and 4 °C for 10 min. SDS-PAGE was used to resolve 30 mg protein per sample. HDAC3 (85057S; 1:1000; Cell Signaling) and GAPDH (A01020; 1:1000; Abbkine) were used as primary antibodies. The samples were incubated with the primary antibodies overnight at 4 °C, followed by incubation with goat anti-rabbit IgG (A21020; 1:5000; Abbkine) or goat anti-mouse IgG antibodies (A21010; 1:5000; Abbkine) for 1 h at 37 °C. Bands were captured using a FluorChem^® Q Imaging System (ProteinSimple, CA, USA) and measured using Image J software (NIH Image, Bethesda, MD, USA). Protein expression was normalised to that of GAPDH.

Zhou L., Wu H., Gao X., Zheng X., Chen H., Li H., Peng J., Liang W., Wang W., Qiu Z., Udduttula A., Wu K., Li L., Liu Y, & Liu Y. (2020). Bone-Targeting Liposome-Encapsulated Salvianic Acid A Improves Nonunion Healing Through the Regulation of HDAC3-Mediated Endochondral Ossification. Drug Design, Development and Therapy, 14, 3519-3533.

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