Rabbit monoclonal antibody against

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Rabbit monoclonal antibody against [target protein]. This antibody is produced in rabbits and can be used to detect the [target protein] in various research applications.

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7 protocols using rabbit monoclonal antibody against

Antibody Panel for Cell Signaling

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Rabbit polyclonal antibody CALR (ab2907), rabbit monoclonal phosphoneoepitope-specific antibody against phospho-eIF2α (Ser 51) (ab32157, clone E90), mouse monoclonal antibody against β-actin (ab49900, clone AC-15) were purchased from Abcam (Cambridge, UK). Rabbit polyclonal antibody against HRI (sc-30143), mouse monoclonal antibody against PKR (sc-6282, clone B-10) were purchased from Santa-Cruz biotechnology. Rabbit monoclonal antibody against PERK (#3192, clone C33E10), rabbit polyclonal antibody against GCN2 (#3302), rabbit polyclonal antibody against eIF2α (#99 722 S), Rabbit monoclonal antibody against LC3B (#2775), rabbit polyclonal antibody against GFP (#2555) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibody anti-PD-1 (BE0273, clone 29F.1A12) and rat IgG1 anti-horseradish peroxidase isotype control (BE0088, clone HRPN) were purchased from BioXcell (West Lebanon, NH, USA). Mouse monoclonal antibody against Atg5 (A2859) was purchased from Merck-Sigma Aldrich. Anti-rabbit Alexa fluor®−488 coupled secondary antibody came from Thermo Fisher Scientific (#A11034). APC anti-mouse CD11c (Clone N418) was purchased from Biolegend (117310).

Bezu L., Wu Chuang A., Sauvat A., Humeau J., Xie W., Cerrato G., Liu P., Zhao L., Zhang S., Le Naour J., Pol J., van Endert P., Kepp O., Barlesi F, & Kroemer G. (2022). Local anesthetics elicit immune-dependent anticancer effects. Journal for Immunotherapy of Cancer, 10(4), e004151.

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Western Blot Analysis of EMT Markers

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Protein lysates were separated by 10% SDS-PAGE, and electrophoretically transferred to PVDF (polyvinylidene difluoride) membrane (Millipore). Then, the membrane was incubated with rabbit monoclonal antibody against human SNAI2, ZEB1, E-cadherin and vimentin (Cell Signaling Technology, USA) followed by HRP (horseradish peroxidase)-labeled goat-anti rabbit IgG (Santa Cruz Biotechnology, USA) and detected by chemiluminescence. GAPDH was used as a protein-loading control.

Liao H., Bai Y., Qiu S., Zheng L., Huang L., Liu T., Wang X., Liu Y., Xu N., Yan X, & Guo H. (2015). MiR-203 downregulation is responsible for chemoresistance in human glioblastoma by promoting epithelial-mesenchymal transition via SNAI2. Oncotarget, 6(11), 8914-8928.

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Leptin Receptor Signaling Pathway Analysis

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Recombinant human leptin was purchased from PeproTech (Rocky Hill, NJ, USA). The rabbit polyclonal antibody against collagen II (ab34712), mouse monoclonal antibody against the leptin receptor (ab2143), and rabbit polyclonal antibody against beta-actin (ab8227) were obtained from Abcam (Cambridge, MA, USA). The chicken anti-rabbit IgG (H+L) secondary antibody (A-21442), chicken anti-mouse IgG (H+L) secondary antibody (A-21200), Sulfo-NHS-SS-Biotin (21331), and avidin agarose (20219) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). The rabbit monoclonal antibody against JAK2, rabbit monoclonal antibody against p-JAK2 (Tyr1008), mouse monoclonal antibody against STAT3, rabbit monoclonal antibody against p-STAT3 (Ser727) and rabbit monoclonal antibody against CHC were purchased from Cell Signaling Technology (Beverly, MA, USA). Toluidine blue (6586-04-5), Safranin O (477-73-6), Alcian blue (33864-99-2), 3-methyladenine (5142-23-4), and MG-132(R) (1211877-36-9) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The Human Leptin ELISA kit was obtained from R&D Systems (Minneapolis, MN, USA).

Wang Y.J., Yu H.G., Zhou Z.H., Guo Q., Wang L.J, & Zhang H.Q. (2016). Leptin Receptor Metabolism Disorder in Primary Chondrocytes from Adolescent Idiopathic Scoliosis Girls. International Journal of Molecular Sciences, 17(7), 1160.

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Western Blot Analysis of Cell Signaling Proteins

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Rabbit polyclonal IgG antibody against human SHP-1 (sc-287) and human β-actin (sc-47778) and a mouse monoclonal IgG antibody against Snail1 (sc-10433) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Mouse monoclonal IgG antibodies against human STAT3 (#9139) and p-STAT3 (Tyr705, #4113), rabbit polyclonal antibodies against human JAK2 (#3230), p-JAK2 (Tyr1007/1008, #3771), E-cadherin (#3195), and a rabbit monoclonal antibody against human vimentin (#5741) were purchased from Cell Signaling Technology (Beverly, MA, USA). A total of 80–100 μg of cell lysate protein was extracted and loaded into SDS-PAGE gel. Primary antibodies were diluted at a ratio of 1:1000 in blocking buffer (Tris-buffered saline with Tween-20; Biosesang, Gyeonggi, Korea) containing 5% skim milk (Difco; Becton–Dickinson and Co., Sparks, MD, USA). Probed membranes were incubated at 4 °C overnight, and then incubated with goat anti-mouse or anti-rabbit IgG as a secondary antibody at room temperature for 1 h. Bands were visualized by exposing membranes to enhanced chemiluminescence (Perkin-Elmer, Waltham, MA, USA) for 1 min.

Koh J.S., Joo M.K., Park J.J., Yoo H.S., Choi B.I., Lee B.J., Chun H.J, & Lee S.W. (2018). Inhibition of STAT3 in gastric cancer: role of pantoprazole as SHP-1 inducer. Cell & Bioscience, 8, 50.

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Investigating TGF-β1 and PPAR-α Regulation

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LPS (isolated from Escherichia coli) and TGF-β1 (used at a dose of 10 µg/ml) were purchased from Sigma-Aldrich; Merck Millipore (Darmstadt, Germany). Mouse monoclonal antibodies against TGF-β1 were from Abcam (cat. no. PB190503; Cambridge, MA, USA) and rabbit monoclonal antibody against PPARα was from Cell Signaling Technology, Inc. (cat. no. 2443S; Beverly, MA, USA). HRP-conjugated anti-mouse and anti-rabbit secondary antibodies were from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Specific small interfering short hairpin (sh)RNA targeting TGF-β1 and PPAR-α and the negative control shRNA were all from Sigma-Aldrich; Merck Millipore. Following allowing cellular attachment to the plates, 2×10⁵ cells were treated with relative shRNAs using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) RNA and protein were collected 72 h following treatment.

Liu Y., Xie L., Yang M., Tan X., Zeng Y., Zheng G., Chen Y, & Chen P. (2017). PPAR-α improves the recovery of lung function following acute respiratory distress syndrome by suppressing the level of TGF-β1. Molecular Medicine Reports, 16(1), 49-56.

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Signaling Pathways in Intestinal Inflammation

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The chemicals were obtained from the following sources: Loperamide hydrochloride (Lop), prucalopride, sodium chloride, activated charcoal, and aqueous methylcellulose from Sigma-Aldrich (St Louis, MO, USA). Rabbit monoclonal antibody against phospho-extracellular signal-regulated kinase (ERK) 1/2 (Thr202/Tyr204), rabbit polyclonal antibodies against phospho-p38 (Thr180/Tyr182) and phospho-SAPK/JNK (Thr183/Tyr185), horseradish peroxidase (HRP)-conjugated anti-rabbit, and anti-mouse immunoglobulin G (IgG) antibodies were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Rabbit anti-c-Kit and anti-SCF antibodies were obtained from Abcam (Cambridge, MA, USA). The BCA protein assay kit was obtained from Thermo Scientific (Waltham, MA, USA). The polyvinylidene difluoride (PVDF) membrane and enhanced chemiluminescence substrate were obtained from Amersham Pharmacia Biotech. The Hybrid-R™ RNA extraction kit was purchased from GeneAll Biotechnology Co., Ltd (Seoul, Korea). The PrimeScript™ 1st strand cDNA Synthesis kit and TB Green^® Premix Ex Taq™ II (Tli RNaseH Plus) were acquired from Takara (Takara, Japan). All other chemicals were of analytical grade or complied with the standards.

Kim H.R., Park I.S., Park S.B., Yang H.J., Jeong D.Y, & Kim S.Y. (2021). Comparison of laxative effects of fermented soybeans (Cheonggukjang) containing toxins and biogenic amines against loperamide-induced constipation mouse model. Nutrition Research and Practice, 16(4), 435-449.

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Quantifying Cellular Stress Responses

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FITC-Dextran 40 (average Mw = 40kDa), recombinant Human TNF-α, and DAPI were obtained from Sigma-Aldrich (St Louis, MO, USA). Lipofectamine® 3000TransfectionReagent, Deoxyribonucleasel, AlexaFluor® 594 Conjugate, and AlexaFluor® 488 Phalloidin were obtained from Invitrogen (Carlsbad, CA, USA). GSTP1 monoclonal antibody andp38monoclonal antibody were obtained from Abcam (Cambridge, United Kingdom). Rabbit polyclonal antibodies against phospho-HSP27 (S15), phospho-HSP27(S78), phospho-HSP27(S82), HSP27, GAPDH, phospho-p38, p38, and Flag-tag were obtained from BioWorld Biotechnology (Minneapolis, MN, USA). Rabbit monoclonal antibody against phospho-MAPK kinaseö (MKK6) was purchased from Cell Signaling Technology (Beverly, MA, USA). Normal mouse IgG, normal rabbit IgG, and TAK1 monoclonal antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Goat anti-rabbit IgG and goat anti-mouse IgG antibodies were purchased from LI-COR Biosciences (Lincoln, NE, USA).

Yang Y., Yin F., Hang Q., Dong X., Chen J., Li L., Cao P., Yin Z, & Luo L. (2018). Regulation of Endothelial Permeability by Glutathione S-Transferase Pi Against Actin Polymerization. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 45(1).

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