
Recombinant Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
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Overview
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Product name
Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free
See all NOTCH4 primary antibodies -
Description
Rabbit monoclonal [EPR18049] to NOTCH4 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt, IP, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment aa 1100-1300. The exact sequence is proprietary.
Database link: Q99466 -
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. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa and NIH/3T3 cells. IP: HeLa whole cell lysate.
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General notes
Ab222400 is the carrier-free version of ab184742. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
ab222400 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
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
Constituent: PBS -
Concentration information loading...
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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
Our Abpromise guarantee covers the use of ab222400 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 |
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Flow Cyt | Use at an assay dependent concentration. | |
IP | Use at an assay dependent concentration. | |
ICC/IF | Use at an assay dependent concentration. | |
WB | Use at an assay dependent concentration. Detects a band of approximately 150 kDa (predicted molecular weight: 210 kDa). |
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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Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling NOTCH4 with ab184742 at 1/200 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on NIH/3T3 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).
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Immunocytochemistry/ Immunofluorescence - Anti-NOTCH4 antibody [EPR18049] - BSA and Azide free (ab222400)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling NOTCH4 with ab184742 at 1/200 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on HeLa cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).
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Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling NOTCH4 with ab184742 at 1/200 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).
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Flow cytometric analysis of 2% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling NOTCH4 with ab184742 at 1/200 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184742).
Protocols
Datasheets and documents
References
ab222400 has not yet been referenced specifically in any publications.