Protein lysates from fibroblasts were prepared using RIPA buffer, separated on Criterion Precast Gels (Bio-Rad) 4–15% Tris-HCl, followed by transfer to PVDF Membrane (Bio-Rad). Membranes were probed with the following primary antibodies in TBS-T and 5% milk for 12 to 16 hours: Anti-Notch1 (R&D AF33471), Anti-Notch3 (V1662) 1ug/ml, GAPDH (1:1000, Cell Signaling). Secondary antibodies (1:10,000) were from Jackson ImmunoResearch. Chemiluminescent detection reagents were from Bio-Rad.
Anti notch1
Manufactured by R&D Systems
Anti-Notch1 is a recombinant protein that binds to and inhibits the Notch1 receptor. Notch1 is a cell surface receptor that plays a key role in cell-cell signaling and cellular differentiation.
Automatically generated - may contain errors
Lab products found in correlation
Criterion precast gel, by Bio-Rad (2 mentions)
Pvdf membrane, by Bio-Rad (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Chemiluminescent detection reagents, by Bio-Rad (2 mentions)
Anti notch3, by Cell Signaling Technology (2 mentions)
Pe mouse igg2a, by BioLegend (1 mentions)
Hrp conjugated secondary antibody, by Merck Group (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Anti notch3, by R&D Systems (1 mentions)
Anti dll4, by R&D Systems (1 mentions)
Recombinant human jagged 1, by R&D Systems (1 mentions)
Anti notch4, by Santa Cruz Biotechnology (1 mentions)
Gsi xii, by Merck Group (1 mentions)
Goat igg pe, by R&D Systems (1 mentions)
Anti jagged1 fitc, by R&D Systems (1 mentions)
Anti dll3 pe, by R&D Systems (1 mentions)
Anti notch1 pe, by BioLegend (1 mentions)
Anti notch2 pe, by BioLegend (1 mentions)
Anti notch3 pe, by BioLegend (1 mentions)
Anti notch4 pe, by BioLegend (1 mentions)
3 protocols using anti notch1
1
Quantitative Notch Signaling Analysis
2
Quantitative Notch Signaling Analysis
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Protein lysates from fibroblasts were prepared using RIPA buffer, separated on Criterion Precast Gels (Bio-Rad) 4–15% Tris-HCl, followed by transfer to PVDF Membrane (Bio-Rad). Membranes were probed with the following primary antibodies in TBS-T and 5% milk for 12 to 16 hours: Anti-Notch1 (R&D AF33471), Anti-Notch3 (V1662) 1ug/ml, GAPDH (1:1000, Cell Signaling). Secondary antibodies (1:10,000) were from Jackson ImmunoResearch. Chemiluminescent detection reagents were from Bio-Rad.
3
Multiparametric Analysis of Notch Signaling in Leukemia
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The antibodies used for FACS analysis were: mouse IgG2b-FITC, goat IgG-PE, anti-Jagged1-FITC, anti-Dll3-PE (all from R&D System, Minneapolis, MN), mouse IgG2a-PE, mouse IgG1κ-PE, mouse IgG1-Alexa Fluor 488,anti-Notch1-PE, anti-Notch2-PE, anti-Notch3-PE, anti-Notch4-PE, anti-Dll1-PE, anti-Dll4, anti-Bax-Alexa Fluor 488 (all from Biolegend, San Diego, CA) and rabbit anti-Bcl-2-FITC (DAKO). For leukemia cell identification we used anti-CD45-VioBlue, anti-CD45-APC-Vio770, anti-CD34-PerCP and anti-CD117-APC (all from MiltenyiBiotec, Germany). The antibodies employed for western blot analysis anti-Notch2, anti-Notch4 were from Santa Cruz (Biotechnology, Dallas, TX), anti-GAPDH and HRP conjugated secondary antibodies against mouse, rabbit or goat were from Sigma Aldrich. All the other antibodies used for Western blot were from Cell Signalling. Neutralizing antibodies,all used at a final concentration of 5 μg/ml, were: anti-Notch1, anti-Notch3,anti-Jagged1, anti-Jagged2, anti-Dll1 and anti-Dll4 (R&D Systems); anti-Notch-4 (Santa Cruz Biotechnology); anti-Dll3 (CST, Boston, MA). Recombinant human Jagged-1 and Jagged-2 were from R&D System. GSI-IX (DAPT) was purchased from Stemgent (Cambridge, MA) GSI-XII and SAHM1 were from Merck Millipore (Darmstadt, Germany). Cytarabine (Ara-C), Etoposide (Eto) and Idarubicin (Ida) were provided by Pharmacy Unit of the University Hospital of Verona.
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