Recombinant Anti-NOTCH4 antibody [EPR18049] (ab184742)
![Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)](https://www.abcam.com/ps/products/184/ab184742/Images/ab184742-267013-anti-notch4-antibody-epr18049-western-blot.jpg "Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18049] to NOTCH4
- Suitable for: IP, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-NOTCH4 antibody [EPR18049]
See all NOTCH4 primary antibodies -
Description
Rabbit monoclonal [EPR18049] to NOTCH4 -
Host species
Rabbit -
Tested applications
Suitable for: IP, WBmore details
Unsuitable for: Flow Cyt (Intra) or ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human NOTCH4 fragment recombinant protein; Human fetal lung, placenta, fetal brain, fetal heart and fetal kidney lysates; HeLa, HepG2, A549, Jurkat, C6, RAW 264.7, PC-12, NIH/3T3 and F9 whole cell lysates; Mouse brain, heart, spleen and placenta lysates; Rat brain and heart lysates. IP: HeLa whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18049 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab184742 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IP |
1/60.
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| WB |
1/1000. Detects a band of approximately 150 kDa (predicted molecular weight: 210 kDa).
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| Notes |
|---|
|
IP
1/60. |
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WB
1/1000. Detects a band of approximately 150 kDa (predicted molecular weight: 210 kDa). |
Application notes
Is unsuitable for Flow Cyt (Intra) or ICC/IF.
Target
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Function
Functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs. May regulate branching morphogenesis in the developing vascular system. -
Tissue specificity
Highly expressed in the heart, moderately in the lung and placenta and at low levels in the liver, skeletal muscle, kidney, pancreas, spleen, lymph node, thymus, bone marrow and fetal liver. No expression was seen in adult brain or peripheral blood leukocytes. -
Sequence similarities
Belongs to the NOTCH family.
Contains 5 ANK repeats.
Contains 28 EGF-like domains.
Contains 3 LNR (Lin/Notch) repeats. -
Post-translational
modificationsSynthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans-Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin dependent gamma-secretase to release a notch-derived peptide containing the intracellular domain (NICD) from the membrane.
Phosphorylated. -
Cellular localization
Cell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4855 Human
- Entrez Gene: 18132 Mouse
- Entrez Gene: 406162 Rat
- Omim: 164951 Human
- SwissProt: Q99466 Human
- SwissProt: P31695 Mouse
- Unigene: 436100 Human
- Unigene: 173813 Mouse
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Alternative names
- FLJ16302 antibody
- hNotch 4 antibody
- hNotch4 antibody
see all
Images
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Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)All lanes : Anti-NOTCH4 antibody [EPR18049] (ab184742) at 1/2000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (Agarose) (ab97052) at 1/100000 dilution
Predicted band size: 210 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,2 and 3: 15 seconds;Lane 4: 3 minute.
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![Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)](https://www.abcam.com/ps/products/184/ab184742/Images/ab184742-267014-anti-notch4-antibody-epr18049-western-blot.jpg)
Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)All lanes : Anti-NOTCH4 antibody [EPR18049] (ab184742) at 1/2000 dilution
Lane 1 : Human fetal lung lysate
Lane 2 : Human placenta lysate
Lane 3 : Human fetal brain lysate
Lane 4 : Human fetal heart lysate
Lane 5 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 210 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,3,4 and 5: 3 minutes; Lane 2: 30 seconds.
The bands beneath the 150 kD NECD band are likely to be the degradation fragments.
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![Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)](https://www.abcam.com/ps/products/184/ab184742/Images/ab184742-267015-anti-notch4-antibody-epr18049-western-blot.jpg)
Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)Anti-NOTCH4 antibody [EPR18049] (ab184742) at 1/1000 dilution + Human NOTCH4 fragment recombinant protein at 0.01 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 210 kDa
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)](https://www.abcam.com/ps/products/184/ab184742/Images/ab184742-267012-anti-notch4-antibody-epr18049-western-blot.jpg)
Western blot - Anti-NOTCH4 antibody [EPR18049] (ab184742)All lanes : Anti-NOTCH4 antibody [EPR18049] (ab184742) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat heart lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 9 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 10 : Mouse placenta lysate
Lane 11 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 210 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,2,3,4 and 5: 1 minute; Lane 6, 7, 8 and 9: 15 seconds; Lane 10 and 11: 30 seconds.
The bands beneath the 150 kD NECD band are likely to be the degradation fragments.
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![Immunoprecipitation - Anti-NOTCH4 antibody [EPR18049] (ab184742)](https://www.abcam.com/ps/products/184/ab184742/Images/ab184742-267007-anti-notch4-antibody-epr18049-immunoprecipitation.jpg)
Immunoprecipitation - Anti-NOTCH4 antibody [EPR18049] (ab184742)NOTCH4 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab184742 at 1/60 dilution.
Western blot was performed from the immunoprecipitate using ab184742 at 1/5000 dilution.
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: HeLa whole cell lysate, 10µg (Input).
Lane 2: ab184742 IP in HeLa whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab184742 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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![Anti-NOTCH4 antibody [EPR18049] (ab184742)](https://www.abcam.com/ps/products/184/ab184742/Images/ab184742-10-benefits-of-recombinant-antibodies.png)
Anti-NOTCH4 antibody [EPR18049] (ab184742)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (13)
ab184742 has been referenced in 13 publications.
- Sharma D et al. IL-33 enhances Jagged1 mediated NOTCH1 intracellular domain (NICD) deubiquitination and pathological angiogenesis in proliferative retinopathy. Commun Biol 5:479 (2022). PubMed: 35589941
- Gao Y et al. Atractylenolide III Attenuates Angiogenesis in Gastric Precancerous Lesions Through the Downregulation of Delta-Like Ligand 4. Front Pharmacol 13:797805 (2022). PubMed: 35846998
- Xia Z et al. Emodin alleviates hypertrophic scar formation by suppressing macrophage polarization and inhibiting the Notch and TGF-ß pathways in macrophages. Braz J Med Biol Res 54:e11184 (2021). PubMed: 34320121
- Torres HM et al. Selective Targeting of Class I Histone Deacetylases in a Model of Human Osteosarcoma. Cancers (Basel) 13:N/A (2021). PubMed: 34439353
- Li G et al. LncRNA PPM1A-AS Regulate Tumor Development Through Multiple Signal Pathways in T-Cell Acute Lymphoblastic Leukemia. Front Oncol 11:761205 (2021). PubMed: 34746000