Anti-NOTCH3 antibody

• Supplier: Abcam
• Catalog: ab23426
• Prices: $401.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: WB IHC-P ICC/IF ICC/IF IHC-F IHC-F ICC/IF
• Species Reactivities: Mouse, Rat, Human
• Antigen: Synthetic peptide corresponding to Human NOTCH3 aa 2300 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH)
• Description: Rabbit Polyclonal
• Gene: NOTCH3
• Conjugate: Unconjugated

Product images

All lanes : Anti-NOTCH3 antibody (ab23426) at 1 µg/mlLane 1 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate - tumor cell line (ab29306)Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate (ab3950)Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L (HRP) at 1/3000 dilutionPerformed under reducing conditions.

ab23426 staining NOTCH3 in mouse embryo tissue (E16.5) by Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue underwent fixation in formaldehyde, heat-mediated antigen retreival in citrate buffer pH6.0 and blocking for 15 minutes at 10°C (5 minutes/peroxidase blocking and 10 minutes/protein blocking). The primary antibody was diluted 1/250 and incubated with sample for 15 minutes at 20°C. A HRP-conjugated goat polyclonal to rabbit IgG was used undiluted as the secondary.See Abreview

ab23426 staining NOTCH3 in human breast cancer tissue sections by immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue underwent fixation in paraformaldehyde, heat-mediated antigen retrieval in citrate buffer pH6.0 and blocking for 15 minutes at 20°C (5 minutes for peroxidase blocking and 10 minutes for protein blocks). The primary antibody was diluted 1/250 and incubated with sample for 45 minutes at 20°C. A HRP-conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.See Abreview

IHC-FoFr image of NOTCH3 staining in Rat cerebral cortex (left) and the paraventricular hypothalamic nucleus (right) sections. The sections used came from animals perfused fixed with Paraformaldehyde 4%, in phosphate buffer 0.2M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Immunostainings were performed using the ‘free floating’ technique. See Abreview

IHC image of NOTCH3 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab23426, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

ICC/IF image of ab23426 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23426, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

ICC/IF image of ab23426 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23426, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

ab14404 staining NOTCH3 in the COS1 fibroblast cell line from Monkey Kkidney by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 0.1% in PBS and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 16 hour at 4°C. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal (1/500) was used as the secondary antibody.See Abreview

Product References

Clinical impact of de-regulated Notch-1 and Notch-3 in the development and - Clinical impact of de-regulated Notch-1 and Notch-3 in the development and

Tripathi R, Rath G, Jawanjal P, Sharma S, Singhal P, Bhambhani S, Hussain S, Bharadwaj M. PLoS One. 2014 Jun 6;9(6):e98642.

Dynamics of Notch pathway expression during mouse testis post-natal development - Dynamics of Notch pathway expression during mouse testis post-natal development

Murta D, Batista M, Silva E, Trindade A, Henrique D, Duarte A, Lopes-da-Costa L. PLoS One. 2013 Aug 28;8(8):e72767.

NOTCH3 is a prognostic factor that promotes glioma cell proliferation, migration - NOTCH3 is a prognostic factor that promotes glioma cell proliferation, migration

Alqudah MA, Agarwal S, Al-Keilani MS, Sibenaller ZA, Ryken TC, Assem M. PLoS One. 2013 Oct 15;8(10):e77299.

Gamma secretase inhibition promotes hypoxic necrosis in mouse pancreatic ductal - Gamma secretase inhibition promotes hypoxic necrosis in mouse pancreatic ductal

Cook N, Frese KK, Bapiro TE, Jacobetz MA, Gopinathan A, Miller JL, Rao SS, Demuth T, Howat WJ, Jodrell DI, Tuveson DA. J Exp Med. 2012 Mar 12;209(3):437-44.

Immunohistochemical study of the epithelial-mesenchymal transition phenotype in - Immunohistochemical study of the epithelial-mesenchymal transition phenotype in

Stoyianni A, Goussia A, Pentheroudakis G, Siozopoulou V, Ioachim E, Krikelis D, Golfinopoulos V, Cervantes A, Bobos M, Fotsis T, Bellou S, Fountzilas G, Malamou-Mitsi V, Pavlidis N. Anticancer Res. 2012 Apr;32(4):1273-81.

Tbx18 function in epicardial development. - Tbx18 function in epicardial development.

Greulich F, Farin HF, Schuster-Gossler K, Kispert A. Cardiovasc Res. 2012 Dec 1;96(3):476-83.

Notch receptor and effector expression in von Hippel-Lindau disease-associated - Notch receptor and effector expression in von Hippel-Lindau disease-associated

Merrill MJ, Edwards NA, Lonser RR. J Neurosurg. 2011 Sep;115(3):512-7.

Asymmetric cell divisions promote Notch-dependent epidermal differentiation. - Asymmetric cell divisions promote Notch-dependent epidermal differentiation.

Williams SE, Beronja S, Pasolli HA, Fuchs E. Nature. 2011 Feb 17;470(7334):353-8.

Modulation of microRNA expression in human T-cell development: targeting of - Modulation of microRNA expression in human T-cell development: targeting of

Ghisi M, Corradin A, Basso K, Frasson C, Serafin V, Mukherjee S, Mussolin L, Ruggero K, Bonanno L, Guffanti A, De Bellis G, Gerosa G, Stellin G, D'Agostino DM, Basso G, Bronte V, Indraccolo S, Amadori A, Zanovello P. Blood. 2011 Jun 30;117(26):7053-62.

Posttranslational stability of the heme biosynthetic enzyme ferrochelatase is - Posttranslational stability of the heme biosynthetic enzyme ferrochelatase is

Crooks DR, Ghosh MC, Haller RG, Tong WH, Rouault TA. Blood. 2010 Jan 28;115(4):860-9.