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Anti hes1

Manufactured by Merck Group
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Anti-Hes1 is a laboratory reagent used to detect and quantify the Hes1 protein, which is a key transcriptional regulator involved in various cellular processes. The core function of Anti-Hes1 is to provide a reliable and specific tool for researchers to analyze Hes1 expression levels in biological samples.

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Lab products found in correlation

Anti jagged1, by Santa Cruz Biotechnology (3 mentions) Anti cleaved notch1, by Cell Signaling Technology (2 mentions) Nitrocellulose membrane, by Merck Group (2 mentions) Anti notch1, by Santa Cruz Biotechnology (2 mentions) Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Enhanced chemiluminescence, by PerkinElmer (2 mentions) Flag m2, by Merck Group (1 mentions) Protease inhibitor, by Beyotime (1 mentions) Anti lamin a c, by Cell Signaling Technology (1 mentions) Anti iκb, by Cell Signaling Technology (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions) Anti p65, by Cell Signaling Technology (1 mentions) Anti p akt, by Cell Signaling Technology (1 mentions) Anti pten, by Cell Signaling Technology (1 mentions) Anti β actin, by Merck Group (1 mentions) Rabbit monoclonal anti notch1 nicd antibody, by Abcam (1 mentions) Mouse mmp7, by R&D Systems (1 mentions) Ab ph 2.5 stain kit, by Agilent Technologies (1 mentions) Anti muc2, by Santa Cruz Biotechnology (1 mentions)

7 protocols using anti hes1

1

Chromatin Immunoprecipitation of Hes-1 Protein

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ChIP was performed according to the manufacturer’s protocol (Millipore). Briefly, nuclear chromatin extracts were incubated with antibody against Hes-1 (Novus, Littleton, CO) (4.8 μg of anti-Hes-1 or mouse IgG) or Flag-M2 (Sigma-Aldrich) at 4°C overnight. Immunoprecipitates were collected on magnesium beads for another 1-2 h at 4°C. After thorough washing, immunoprecipitates were de-crosslinked and chromatin was recovered for quantitative PCR analysis. Primers used for GADD45α promoter are: 5′-TCATGATTCAGCATCTAACATCAATAA-3′ (forward) and 5′-GACAACCATCTGACACCC-3′ (reverse).
Chiou H.Y., Liu S.Y., Lin C.H, & Lee E.H. (2014). Hes-1 SUMOylation by protein inhibitor of activated STAT1 enhances the suppressing effect of Hes-1 on GADD45α expression to increase cell survival. Journal of Biomedical Science, 21(1), 53.
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2

Western Blot Analysis of BM-Derived Macrophages

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BM-derived macrophages were harvested and lysed on ice for 30 min in RIPA buffer supplemented with protease inhibitors (Beyotime, Haimen, China), and lysates then centrifuged at 12,000 rpm for 10 min. The supernatants were collected, and protein concentrations were determined using BCA Protein Assay reagents (Pierce, Waltham, MA, USA). Nuclei were isolated using a kit (Beyotime) according to the supplier’s protocol and lysed as described above. Aliquots of cell lysates were separated by SDS-PAGE and then blotted onto PVDF membranes. Membranes were blocked with 5% skim milk solution for 2 h and then probed with primary antibodies, followed by horseradish peroxidase-conjugated goat anti-rabbit IgG (1:2,500) or goat anti-mouse IgG (1:2,000) (Boster Bio Tec, Wuhan, China). Protein blots were visualized using an ECL detection system (Pierce). The following primary antibodies were used: anti-PTEN, anti-Akt, anti-p-Akt, anti-IκB, anti-p65 and anti-Lamin A/C (Cell Signaling, Boston, MA, USA), anti-FIH1 (Abgent, San Diego, CA, USA), anti-HES1, and anti-β-actin (Sigma).
Huang F., Zhao J.L., Wang L., Gao C.C., Liang S.Q., An D.J., Bai J., Chen Y., Han H, & Qin H.Y. (2017). miR-148a-3p Mediates Notch Signaling to Promote the Differentiation and M1 Activation of Macrophages. Frontiers in Immunology, 8, 1327.
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3

Western Blotting of Notch Signaling

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Western blotting was performed with standard methods. Briefly, 25 to 50 μg of proteins extracted from cultured cells was separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes (Millipore Co). The membranes were blocked with milk or bovine serum albumin and blotted with relevant antibodies. Horseradish peroxidase–conjugated secondary antibodies were detected by enhanced chemiluminescence (PerkinElmer, Waltham, Mass). Nuclear and cytoplasmic portions were extracted with the NE‐PER nuclear and cytoplasmic extraction kit (Thermo Scientific, Rockford, Ill) according to the manufacturer's instructions. A commercially available antibody used for the Western blot analysis of anti–cleaved Notch1 was purchased from Cell Signaling Technology (Danvers, Mass); anti‐Notch1, anti‐Jagged1, β‐actin, and secondary antibodies were acquired from Santa Cruz Biotechnology (Santa Cruz, Calif); and anti‐Hes1 was obtained from Millipore (Billerica, Mass).
Shang H., Braggio D., Lee Y., Al Sannaa G.A., Creighton C.J., Bolshakov S., Lazar A.J., Lev D, & Pollock R.E. (2015). Targeting the notch pathway: A potential therapeutic approach for desmoid tumors. Cancer, 121(22), 4088-4096.
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4

Comprehensive Immunohistochemistry Analysis

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Tissue was processed for routine IHC staining using the following antibodies: rabbit anti-Lysozyme (Diagnostic BioSystems, Pleasanton, CA, USA; RP 028-05), rabbit monoclonal anti-Notch1 (NICD) antibody (Epitomics Inc. 1935-1), anti-Hes1 (Millipore, 5702), anti-MUC2 (Santa Cruz, SC15334), anti-phospho-S6 (pS235/236) (Cell Signaling, #4858), rabbit chromogranin A (Abcam, Cambridge, MA, USA; 15160), mouse MMP7 (R&D Systems, Minneapolis, MN, USA; AF2967) and rabbit Ki67 (Novus, Littleton, CO, USA; NB110-89717). Negative controls (including no primary antibody or isotype-matched mouse immunoglobulin G) were used in each assessment. AB staining was performed according to standard protocol using AB pH 2.5 Stain Kit (Dako, Carpinteria, CA, USA; AR160). Apoptotic cells were detected by TUNEL assay using an ApopTag Peroxidase In Situ Apoptosis Detection Kit (Millipore, S7100).
Zhou Y., Rychahou P., Wang Q., Weiss H.L, & Evers B.M. (2015). TSC2/mTORC1 signaling controls Paneth and goblet cell differentiation in the intestinal epithelium. Cell Death & Disease, 6(2), e1631-.
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5

Immunoblotting Analysis of Neural Stem Cell Markers

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Anti-Sox2 (goat polyclonal, 1/1,000 dilution, R&D systems), anti-Nestin (mouse monoclonal, 1/1,000 dilution, BD), anti-GFAP (mouse monoclonal, 1/1,000 dilution, ImmunO), anti-NICD (rabbit polyclonal, 1/1,000, Cell Signaling), anti-HES1 (rabbit polyclonal, 1/1,000 dilution, Millipore), anti-Tuj1 (mouse monoclonal, 1/1,000 dilution, Abcam), anti-Symplekin (clone 25, mouse monoclonal, 1:1,000 dilution, BD), and anti-β-Actin (clone C4, mouse monoclonal, 1/1,000 dilution, Santa Cruz Biotech), anti-FLAG (clone M2, mouse monoclonal, 1/2,000 dilution, Sigma-Aldrich; rabbit polyclonal, 1/1,000 dilution, Cell Signaling), anti-HA (clone 3F10, rat monoclonal, 1/1,000 dilution, Roche), and anti-GFP (B-2, mouse monoclonal, 1/1,000 dilution, Santa Cruz Biotech) antibodies were used through the all WB analysis. As a secondary antibody, horseradish peroxidase-conjugated anti-rabbit (1/5,000 dilution, Vector Laboratories), anti-mouse IgG (1/5,000 dilution, Vector Laboratories), and anti-rat IgG (1/5,000 dilution, Santa Cruz Biotech) were used [57 (link), 58 (link)].
Yin J., Park G., Lee J.E., Park J.Y., Kim T.H., Kim Y.J., Lee S.H., Yoo H., Kim J.H, & Park J.B. (2014). CPEB1 modulates differentiation of glioma stem cells via downregulation of HES1 and SIRT1 expression. Oncotarget, 5(16), 6756-6769.
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6

Notch Signaling Pathway Protein Analysis

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Western blot was done by standard methods. Briefly, 25 to 50 μg of proteins extracted from cultured cells were separated by SDS-PAGE and transferred onto nitrocellulose membranes (Millipore Co.). Membranes were blocked with milk or BSA and blotted with relevant antibodies. HRP conjugated secondary antibodies were detected by enhanced chemiluminescence (Perkinelmer, Waltham, MA). Nuclear and Cytoplasmic portions were extracted with NE-PER* Nuclear and Cytoplasmic Extraction Kit (Thermo Scientific, Rockford, IL) according to the manufacturer's instructions. Commercially available antibody used for western blot analysis of anti-cleaved Notch1 was purchased from Cell Signaling (Cell Signaling Technology, Danvers, MA); anti-Notch1, anti-Jagged1, β-actin and secondary antibodies were acquired from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Santa Cruz, CA); anti-Hes1 was obtained from Millipore (Millipore, Billerica, MA).
Shang H., Braggio D., Lee Y., Al Sannaa G.A., Creighton C.J., Bolshakov S., Lazar A.J., Lev D, & Pollock R.E. (2015). Targeting the notch pathway: A potential therapeutic approach for desmoid tumors. Cancer, 121(22), 4088-4096.
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7

Immunohistochemical Profiling of CNS Markers

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For immunohistochemical, immunocytochemical, and Western blot procedures please reference the Supplemental Experimental Procedures. Antibodies used for immunohistochemistry were anti-GFP (Abcam, 1:500), anti-Brdu (Accurate,1:200, 30 min 2N HCl followed by 15 min 0.1M Boric Acid brain section pre-treatment), anti-NG2 (Millipore, 1:500), anti-GFAP (Sigma, 1:500), anti-ET-1 (Abbiotec, 1:200), anti-CD31 (BD Biosciences, 1:500), anti-Jagged-1 (Iowa Hybridoma Bank, 1:200), anti-IBA1 (Wako, 1:500), anti-MAG (Santa Cruz, 1:200), anti-MBP (Covance (SMI-99p), 1:1000), anti-Hes1 (Millipore, 1:1000), anti-CD11b/MAC1 (ABD Serotec, 1:400), anti-Olig2 (Millipore, 1:500), and anti-APC (Ab-7) (CC-1) (Calbiochem, 1:500). Antibodies used for immunocytochemistry were anti-GFP (Abcam, 1:500), anti-O1 (R&D systems, 1:500), anti-GFAP (Sigma, 1:500), and anti-NG2 (Millipore, 1:500). Antibodies used for Western Blot analysis include anti-MBP (Covance (SMI-99p), 1:5000), anti-MAG (Santa Cruz (sc-15324), 1:200), anti-CNPase (Covance, 1:500), anti-Jagged1 (Santa Cruz (sc-135955), 1:200), anti-β-actin(C4) (Millipore, 1:5000), anti-GFAP (Sigma, 1:5000), and anti-NICD (Iowa Hybridoma Bank C17.9C6, 1:1000).
Hammond T.R., Gadea A., Dupree J., Kerninon C., Nait-Oumesmar B., Aguirre A, & Gallo V. (2014). Astrocyte-derived Endothelin-1 inhibits remyelination through Notch activation. Neuron, 81(3), 588-602.
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