Name: Anti-Notch1 antibody [EP1238Y]
SKU: ab52627
Rating: 5 (17 reviews)

Anti-Notch1 antibody EP1238Y is a rabbit monoclonal antibody that is used in Notch1 Western blot, IHC, ICC/IF, Flow cytometry, IP. Suitable for Human, Mouse samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with NOTCH1 knockout cell line validation
- Specificity and sensitivity confirmed in immunohistochemistry (IHC) with multi-tissue microarray (TMA) validation
- Cited in over 200 publications

Related conjugates and formulations

ab194122
Conjugated
Alexa Fluor® 647 Anti-Notch1 antibody [EP1238Y]
ab303042
Conjugated
APC Anti-Notch1 antibody [EP1238Y]
ab303043
Conjugated
HRP Anti-Notch1 antibody [EP1238Y]
ab308809
Conjugated
AP Anti-Notch1 antibody [EP1238Y]
ab310540
Conjugated
Alexa Fluor® 594 Anti-Notch1 antibody [EP1238Y]
ab309758
Conjugated
Alexa Fluor® 488 Anti-Notch1 antibody [EP1238Y]
ab221603
Carrier free
Anti-Notch1 antibody [EP1238Y] - BSA and Azide free
ab303041
Conjugated
PE Anti-Notch1 antibody [EP1238Y]
ab315420
Conjugated
Alexa Fluor® 555 Anti-Notch1 antibody [EP1238Y]
ab321656
Conjugated
Alexa Fluor® 750 Anti-Notch1 antibody [EP1238Y]

Images

Western blot - Anti-Notch1 antibody [EP1238Y] (AB52627), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Not recommended	Tested	Tested	Tested
Mouse	Expected	Not recommended	Tested	Expected	Expected
Cow	Predicted	Not recommended	Predicted	Predicted	Predicted

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/150	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human, Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/2000	Notes -
Species Human	Dilution info 1/1000 - 1/2000	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/150	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/130 - 1/200	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Cow	Dilution info -	Notes -

Associated Products

Select an associated product type

4 products for Alternative Product

1 product for Alternative Version

Target data

See full target information NOTCH1

Function

Functions as a receptor for membrane-bound ligands Jagged-1 (JAG1), Jagged-2 (JAG2) and Delta-1 (DLL1) 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. Involved in angiogenesis; negatively regulates endothelial cell proliferation and migration and angiogenic sprouting. Involved in the maturation of both CD4(+) and CD8(+) cells in the thymus. Important for follicular differentiation and possibly cell fate selection within the follicle. During cerebellar development, functions as a receptor for neuronal DNER and is involved in the differentiation of Bergmann glia. Represses neuronal and myogenic differentiation. May play an essential role in postimplantation development, probably in some aspect of cell specification and/or differentiation. May be involved in mesoderm development, somite formation and neurogenesis. May enhance HIF1A function by sequestering HIF1AN away from HIF1A. Required for the THBS4 function in regulating protective astrogenesis from the subventricular zone (SVZ) niche after injury. Involved in determination of left/right symmetry by modulating the balance between motile and immotile (sensory) cilia at the left-right organiser (LRO).

Alternative names

TAN1, NOTCH1, Neurogenic locus notch homolog protein 1, Notch 1, hN1, Translocation-associated notch protein TAN-1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EP1238Y
Purification technique: Affinity purification Protein A
Specificity: 80% identities with Notch 2 and 81% with Notch 3
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Notch 1 (encoded by the Notch 1 gene) is a member of the Notch family of receptors. When bound to a ligand, the Notch1 receptor sends signals critical for normal development of many tissues throughout the body. Notch1 signalling supports the determination and specialization of cells as well as playing a role in cell proliferation, differentiation, and apoptosis.

Product Specifications
Anti-Notch1 antibody [EP1238Y] (ab52627) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P, WB in human, mouse samples.
Anti-Notch1 antibody [EP1238Y] (ab52627) specifically detects Notch1 (UniProt ID: P46531; Molecular weight: 271kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-Notch1 antibody [EP1238Y] (ab52627) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-Notch1 antibody [EP1238Y] (ab52627) has been confirmed by testing in knockout samples.
Anti-Notch1 antibody [EP1238Y] (ab52627) has been cited over 201 times in peer reviewed journals and is trusted by the scientific community.
Anti-Notch1 antibody [EP1238Y] (ab52627) has 17 independent reviews from customers.

Related Products
Antibody clone EP1238Y is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor® 647, PE, APC, HRP, Alkaline Phosphatase, Alexa Fluor® 488, Alexa Fluor® 594, Alexa Fluor® 555, Alexa Fluor® 750 (ab194122, ab303041, ab303042, ab303043, ab308809, ab309758, ab310540, ab315420, ab321656).

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 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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
Western blot: Anti-NOTCH1 antibody [EP1238Y] (ab52627) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52627 was shown to bind specifically to NOTCH1. A band was observed at 110-120 kDa in wild-type HL-60 cell lysates with no signal observed at this size in NOTCH1 knockout cell line. To generate this image, wild-type and NOTCH1 knockout HL-60 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution
Lane 1: Wild-type HL-60 cell lysate at 20 µg
Lane 2: NOTCH1 knockout HL-60 cell lysate at 20 µg
Lane 3: Wild-type HAP1 cell lysate at 20 µg
Lane 4: NOTCH1 knockout HAP1 cell lysate at 20 µg
Lane 5: MOLT-4 cell lysate at 20 µg
Lane 6: HepG2 cell lysate at 20 µg
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 110-120 kDa
Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
Lanes 1- 2: Merged signal (red and green). Green - ab52627 observed at 110-120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab52627 was shown to react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human NOTCH1 knockout HeLa cell line ab261762 (knockout cell lysate Human NOTCH1 knockout HeLa cell lysate ab257006) was used. Wild-type HeLa and NOTCH1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab52627 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NOTCH1 knockout HeLa cell line (Human NOTCH1 knockout HeLa cell line ab261762)
Lane 2: NOTCH1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 272 kDa
Observed band size: 110-120 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)
Immunohistochemical staining of paraffin-embedded human brain with purified ab52627 at a dilution of 1/150.
A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution
All lanes: Mouse brain at 10 µg
Secondary
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 272 kDa
Observed band size: 110-120 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)
Verification of gene expression array data by immunohistochemical analysis of Notch 1 expression in subcutaneous tumors and lung metastases from a human melanoma (MeWo) xenograft experiment in mice.
Immunohistochemical staining for Notch1 expression (ab52627, red) in subcutaneous tumors and lung metastases (both panels) of MeWo (Human malignant melanoma cell line) cells.
All scale bars: 50 μm.
Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/2000 dilution
All lanes: HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg
Secondary
All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 272 kDa
Observed band size: 110-120 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (ab52627)
Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using purified ab52627 at a dilution of 1/150.
An Alexa Fluor^® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor^® 555 goat anti-rabbit was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.
Flow Cytometry (Intracellular) - Anti-Notch1 antibody [EP1238Y] (ab52627)
Intracellular Flow Cytometry analysis of permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (2% PFA, pink) withpurified ab52627 at a 1/200 dilution, or negative control rabbit monoclonal IgG (green). The secondary antibody was FITC goat anti-rabbit.
Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627)
Lanes 1 - 4: Merged signal (red and green). Green - ab52627 observed at 110-120 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab52627 was shown to react with Notch1 in wild-type HAP1 cells in western blot. Loss of signal was observed when NOTCH1 knockout sample was used. Wild-type and NOTCH1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab52627 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Notch1 antibody [EP1238Y] (ab52627) at 1 µg/mL
Lane 1: Wild-type HAP1 cell lysate at 40 µg
Lane 2: NOTCH1 knockout HAP1 cell lysate at 40 µg
Lane 3: MOLT-4 cell lysate at 20 µg
Lane 4: HepG2 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 272 kDa
Observed band size: 110-120 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] (ab52627)
Immunohistochemical staining of paraffin-embedded human brain with unpurified ab52627 at a dilution of 1/100.
A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] (ab52627)
Immunofluorescent staining of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells fixed with 4% PFA using unpurified ab52627 at a dilution of 1/100.
An Alexa Fluor^® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor^® 555 goat anti-rabbit was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.