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Jagged 1

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Jagged-1 is a cell surface receptor protein that plays a key role in the Notch signaling pathway. It functions as a ligand, binding to and activating the Notch receptor, which initiates a cascade of intracellular signaling events. Jagged-1 is involved in various cellular processes, including cell fate determination, differentiation, and development.

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Lab products found in correlation

Notch1, by Cell Signaling Technology (4 mentions) E cadherin, by Cell Signaling Technology (3 mentions) β actin, by Cell Signaling Technology (3 mentions) Jagged 2, by Cell Signaling Technology (2 mentions) Notch2, by Cell Signaling Technology (2 mentions) N cadherin, by Cell Signaling Technology (2 mentions) Nicastrin, by Cell Signaling Technology (2 mentions) Las 4000, by Cytiva (2 mentions) β catenin, by Cell Signaling Technology (2 mentions) Snail, by Cell Signaling Technology (2 mentions) Alexa fluor dye, by Thermo Fisher Scientific (2 mentions) α sma, by Merck Group (2 mentions) Eclipse e800 microscope, by Nikon (2 mentions) Evos xl core, by Thermo Fisher Scientific (2 mentions) Metavue research imaging system, by Molecular Devices (2 mentions) Jagged 1, by PerkinElmer (2 mentions) Slowfade gold antifade mount with dapi, by Thermo Fisher Scientific (2 mentions) Erk1 2, by Cell Signaling Technology (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) α sma, by Abcam (1 mentions)

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Jagged-1-#2620 (2 citations)

20 protocols using jagged 1

1

Antibody Validation for Signaling Pathways

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Antibodies to H3K27me3, JMJD3, p-Smad3, Smad3, p-AKT, AKT, p-ERK1/2, ERK1/2, Notch1, Jagged-1, UTX and DNMT1 were purchased from Cell Signaling Technology (MA, USA). Antibodies to fibronectin, α-SMA, collagen III, JMJD3, TGF-β1, Notch3 were purchased from Abcam (MA, USA). An antibody to Smad7 was obtained from Santa Cruz Biotechnology (CA, USA). An antibody to β-actin, PTEN and FBXW7 were obtained from proteintech (Wuhan, China). β-tubulin was purchased from Sigma-Aldrich (MO, USA). GSKJ4 was purchased from Selleck (Houston, USA). DMSO and other chemicals were obtained from Beyotime (Shanghai, China). Lipofectamine 3000 was obtained from Invitrogen-Thermo Fisher Scientific (CA, USA).
Yu C., Xiong C., Tang J., Hou X., Liu N., Bayliss G, & Zhuang S. (2021). Histone demethylase JMJD3 protects against renal fibrosis by suppressing TGFβ and Notch signaling and preserving PTEN expression. Theranostics, 11(6), 2706-2721.
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2

Notch Pathway and Epithelial-Mesenchymal Transition Antibodies

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Polyclonal antibodies to Notch1, Notch2, Notch3, Notch4, Dll1, Dll3, Dll4, Jagged1, Jagged2, HIF‐1α, N‐cadherin, E‐cadherin and Snail1 were purchased from Cell Signaling Technology (Beverly, MA). Polyclonal antibodies to G6PD, transketolase (TKT), Nrf2, Keap1 and monoclonal antibody to β‐actin were purchased from Abcam (Cambridge, UK). The dual‐luciferase reporter assay kit was obtained from Promega (Madison, WI).
Zhang H., Zhang Z., Du G., Sun H., Liu H., Zhou Z., Gou X., Wu X., Yu X, & Huang Y. (2019). Nrf2 promotes breast cancer cell migration via up‐regulation of G6PD/HIF‐1α/Notch1 axis. Journal of Cellular and Molecular Medicine, 23(5), 3451-3463.
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3

Notch Signaling Pathway Regulation

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ZER (purity >98%) was purchased from Sigma-Aldrich (St. Louis, MO). Reagents necessary for cell culture, including fetal bovine serum and antibiotics, and Oligofectamine were purchased from Invitrogen-Life Technologies (Carlsbad, CA). Antibodies for detection of cleaved Notch1, Jagged1, Jagged2, Notch2, cleaved poly-(ADP-ribose)-polymerase (PARP), cleaved caspase-3, Bcl-2, Presenilin-1, and Nicastrin were from Cell Signaling Technology (Beverly, MA); an antibody specific against cleaved Notch2 was from EMD-Millipore (Billerica, MA); anti- Notch4 (detects both full length and cleaved form) antibody was from Santa Cruz Biotechnology (Dallas, TX); an antibody specific for immunodetection of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was from GeneTex (Irvine, CA); and anti-actin antibody was from Sigma-Aldrich. The Notch2 and Presinilin1-targeted small interfering RNA (siRNA), Notch2-targeted small hairpin RNA (shRNA), and control shRNA were purchased from Santa Cruz Biotechnology.. A nonspecific control siRNA was purchased from Qiagen (Germantown, MD). MCF-7 and MDA-MB-231 cells were purchased from American Type Culture Collection (Manassas, VA) and maintained as described previously [27 (link)]. SUM159 cells (Asterand, Detroit, MI) were cultured as suggested by the supplier.
Sehrawat A., Sakao K, & Singh S.V. (2014). Notch2 activation is protective against anticancer effects of zerumbone in human breast cancer cells. Breast cancer research and treatment, 146(3), 543-555.
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4

Protein Expression Analysis of Notch Signaling

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The concentration of extracted protein was detected by Protein BCA Assay (Beyotime, Jiangsu, China). Protein was separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes. After blocking with nonfat milk, primary antibodies specific for Jagged-1 (Cell Signaling Technology, CST, Boston, USA), full-length Notch-1 (CST), cleaved Notch-1 (NICD, Abcam, Cambridge, UK), Synaptopodin (Abcam), Hes-1 (Abcam), VEGF (Abcam), p53 (CST), p62 (Abcam), LC3B (Abcam) were used. Goat anti-rabbit IgG conjugated with horseradish peroxidase was used to detect the expression of primary antibodies, and protein bands were visualized by ECL western blotting detection system (Amersham, Little Chalfont, UK).
He Y., Zhang M., Wu Y., Jiang H., Fu H., Cai Y., Xu Z., Liu C., Chen B, & Yang T. (2018). Aberrant activation of Notch-1 signaling inhibits podocyte restoration after islet transplantation in a rat model of diabetic nephropathy. Cell Death & Disease, 9(10), 950.
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5

Jagged-1 Immunoprecipitation Protocol

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Jagged-1(Jagged 1 2H28 rabbit monoclonal antibody, Cat. 2620, Cell Signaling) was immunoprecipitated under treatment conditions as described in the figure legends. Lysates at a concentration of 3–5mg/mL were incubated with 6μg of antibody for the specific protein of interest or rabbit isotype control IgG (Cat. SC-2027, Santa Cruz, Santa Cruz, CA, USA) overnight with gentle rocking at 4°C. Thirty microliters of protein A-plus beads (sc-2002, Santa Cruz, CA, USA) were added to the immune complexes for two hours, the beads were washed with lysis buffer and the proteins attached to the pelleted beads were eluted with 20μl of 2X Laemmli sample buffer plus β-mercaptoethanol and heated for 10 minutes at 95°C while vigorously shaking. Immunoblots are described in supplemental section.
Pandya K., Wyatt D., Gallagher B., Shah D., Baker A., Bloodworth J., Zlobin A., Pannuti A., Green A., Ellis I.O., Filipovic A., Sagert J., Rana A., Albain K.S., Miele L., Denning M.F, & Osipo C. (2015). PKCα Attenuates Jagged-1-mediated Notch Signaling in ErbB-2 positive Breast Cancer to Reverse Trastuzumab Resistance. Clinical cancer research : an official journal of the American Association for Cancer Research, 22(1), 175-186.
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6

Western Blotting Analysis of Angiogenic Markers

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Western blotting experiments were performed according to previous studies [40 (link)]. Briefly, proteins (50 μg) were loaded into a 10% SDS-PAGE gel and electrotransferred to a polyvinylidene difluoride membrane (Millipore, USA). Membranes were incubated overnight at 4°C in primary antibodies recognizing Jagged-1 (1 : 1000, Cell Signaling Technology), Notch-1 (1 : 1000, Cell Signaling Technology), Hes-1 (1 : 1000, Cell Signaling Technology), CD31 (1 : 1000 dilution, Abcam), zonula occludens-1 (ZO-1) (1 : 1000 dilution, Abcam), and vWF (1 : 500, Santa Cruz Biotechnology). Membranes were washed in TBS and incubated in secondary antibodies conjugated to horseradish peroxidase (HRP) (1 : 1000, Cell Signaling Technology) for 2 h at room temperature. GAPDH (1 : 1000 dilution, Abcam) was used as an endogenous loading control. The ImageQuant LAS 4000 mini detection system (GE Healthcare, Buckinghamshire, UK) was used for quantified densitometric analysis, and ImageJ software (National Institutes of Health, Bethesda, MD) was used to analyze results.
Li X., Luo D., Hou Y., Hou Y., Chen S., Zhan J., Luan J., Wang L, & Lin D. (2020). Sodium Tanshinone IIA Silate Exerts Microcirculation Protective Effects against Spinal Cord Injury In Vitro and In Vivo. Oxidative Medicine and Cellular Longevity, 2020, 3949575.
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7

Immunoblotting Antibodies for EMT and Stemness

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The following antibodies were used for immunoblotting: the antibodies against E-cadherin (Cell Signaling), vimentin (Cell signaling), ZO1 (Cell signaling), Claudin-1 (Cell Signaling), β-catenin (Cell Signaling), ZEB1 (Cell Signaling), Slug (Cell Signaling), Snail (Cell Signaling), p-STAT3 (Cell Signaling), STAT3 (Cell Signaling), p-AKT (Cell signaling), AKT (Cell signaling), p-ERK (Santa Cruz), ERK (Santa Cruz), CD133 (Abcam), OCT4 (Cell signaling), Jagged-1 (Cell signaling), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Sungene Biotech).
Yang Z., Guo L., Liu D., Sun L., Chen H., Deng Q., Liu Y., Yu M., Ma Y., Guo N, & Shi M. (2014). Acquisition of resistance to trastuzumab in gastric cancer cells is associated with activation of IL-6/STAT3/Jagged-1/Notch positive feedback loop. Oncotarget, 6(7), 5072-5087.
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8

Notch Signaling Pathway Analysis

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GSI was provided by Pfizer (New York, New York). Paclitaxel was purchased from the MD Anderson pharmacy. Notch3, Jagged-1, RPS6KB1, DNM1–3, control siRNAs, and Dynasore were purchased from Sigma-Aldrich (St. Louis, Missouri). Primers included PSEN1, APH1A, NCSTN, APH1B, PSENEN, RPS6KB1, DNM1–3, and 18s and were also purchased from Sigma-Aldrich. Antibodies used in this study included Jagged-1, Jagged-2, DLL1, DLL4, P70S6K (RPS6KB1), Notch1–4, and cleaved Notchs and were obtained from Cell Signaling Technology (Danvers, Massachusetts). Anti-DLL3 antibody was obtained from Abcam. β-actin was purchased from Sigma-Aldrich. CD31 was purchased from BD Biosciences (San Jose, California). Anti-Ki67 antibody was purchased from Neomarkers (Fremont, California). Antibody against cleaved caspase-3 was purchased from Cell Signaling Technology. Horseradish peroxidase-conjugated rat anti-mouse IgG2a was purchased from Harlan Bioproducts for Science (Indianapolis, Indiana).
Hu W., Liu T., Ivan C., Sun Y., Huang J., Mangala L.S., Miyake T., Dalton H.J., Pradeep S., Rupaimoole R., Previs R.A., Han H.D., Bottsford-Miller J., Zand B., Kang Y., Pecot C.V., Nick A.M., Wu S.Y., Lee J.S., Sehgal V., Ram P., Liu J., Tucker S.L., Lopez-Berestein G., Baggerly K.A., Coleman R.L, & Sood A.K. (2014). Notch3 Pathway Alterations in Ovarian Cancer. Cancer research, 74(12), 3282-3293.
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9

Protein Expression Analysis in Bone Tissue

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For protein analysis, tissues isolated from the drill hole region were used and crushed in liquid nitrogen. Estimation of protein concentration was quantitated by Bradford assay. 40 micrograms of total protein was then resolved by 10% SDS-PAGE gel. After transfer of proteins onto PVDF membranes (Immobilon-P, Millipore, Billerica, MA, USA), blocking was done with 1% BSA. The membranes were probed with β-Catenin, Phospho β-Catenin(ser 33/37/Thr 41), GSK-3β, LEF-1,smad 1, psmad1/5/8, Ihh, Smo, Notch 1, Jagged 1, c-myc, Hes-1, p21 and β-Actin antibodies (Cell Signaling Technology, Danvers, MA, USA) followed by incubation with HRP conjugated secondary antibodies (Cell Signaling Technology, Cell Signaling Technology, Danvers, MA, USA). Membranes were developed using an enhanced chemiluminescence kit (Millipore, Billerica, MA, USA) using Image Quant LAS 4000 (GE Healthcare, Little Chalfont, UK).
Dixit M., Raghuvanshi A., Gupta C.P., Kureel J., Mansoori M.N., Shukla P., John A.A., Singh K., Purohit D., Awasthi P., Singh D, & Goel A. (2015). Medicarpin, a Natural Pterocarpan, Heals Cortical Bone Defect by Activation of Notch and Wnt Canonical Signaling Pathways. PLoS ONE, 10(12), e0144541.
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10

Notch Signaling Pathway Analysis

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Antibodies against Notch1, Notch2, Jagged1, Hes1, Nicastrin, Dll4, E‐Cadherin, N‐cadherin, Slug, Snail and β‐actin were obtained from Cell Signaling Technology (Danvers, MA). All other chemicals were purchased from Fisher Scientific (Suwanee, GA) and Sigma‐Aldrich (St. Louis, MO).
Chandra Boinpelly V., Verma R.K., Srivastav S., Srivastava R.K, & Shankar S. (2020). α‐Mangostin‐encapsulated PLGA nanoparticles inhibit colorectal cancer growth by inhibiting Notch pathway. Journal of Cellular and Molecular Medicine, 24(19), 11343-11354.
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