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AB300528

Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free)

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Rabbit Recombinant Monoclonal NOTCH3 antibody. Carrier free. Suitable for WB, IHC-P, Flow Cyt (Intra) and reacts with Human samples.

View Alternative Names

Neurogenic locus notch homolog protein 3, Notch 3, NOTCH3

8 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Panel A HEK-293T c tissue labeling NOTCH3 with ab300527 at 1/1000 (0.506 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on (A) HEK-293T transfected with a human NOTCH3 expression vector, no staining on (B) HEK-293T transfected with empty vector. The section was incubated with ab300527 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Flow Cytometry (Intracellular) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.Flow cytometric analysis of MDA-MB-231 (human breast adenocarcinoma epithelial cell, Left) / MCF7 (human breast adenocarcinoma epithelial cell, Right) cells labelling NOTCH3 with ab300527 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression : MDA-MB-231(PMID : 29109797).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human smooth muscle tissue labeling NOTCH3 with ab300527 at 1/500 (1.012 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on vascular smooth muscle cells in human smooth muscle. The section was incubated with ab300527 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling NOTCH3 with ab300527 at 1/500 (1.012 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human lung carcinoma. The section was incubated with ab300527 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling NOTCH3 with ab300527 at 1/500 (1.012 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on vascular smooth muscle cells of human enodmetrium. The section was incubated with ab300527 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • WB

Lab

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (<a href='/en-us/products/primary-antibodies/notch3-antibody-epr25183-101-ab300527'>ab300527</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate (frozen) at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate (fresh) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 90 kDa,270 kDa

false

Exposure time: 180s

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • WB

Supplier Data

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (<a href='/en-us/products/primary-antibodies/notch3-antibody-epr25183-101-ab300527'>ab300527</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeti NOTCH3, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 90 kDa,270 kDa

true

Exposure time: 169s

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)
  • WB

Supplier Data

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (BSA and Azide free) (AB300528)

This data was developed using ab300527, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-NOTCH3 antibody [EPR25183-101] (<a href='/en-us/products/primary-antibodies/notch3-antibody-epr25183-101-ab300527'>ab300527</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate

Lane 2:

MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 90 kDa,270 kDa

true

Exposure time: 48s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR25183-101

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

IHC-P, WB, Flow Cyt (Intra)

applications

Immunogen

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

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Complementary Products

Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Primary Antibodies

AB52627

Anti-Notch1 antibody [EP1238Y]

RabMAb|Recombinant|KO Validated|20ul selling size|Advanced Validation

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

  • 5
  • 18
Primary Antibodies

AB9485

Anti-GAPDH antibody - Loading Control

BOND RX™ Validated|Lab Essentials

Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (AB9485)

  • 5
  • 88
Primary Antibodies

AB307701

Anti-Notch2 antibody [EPR26111-69] - BSA and Azide free

RabMAb|Recombinant

Western blot - Anti-Notch2 antibody [EPR26111-69] - BSA and Azide free (AB307701)

  • 0
  • 0
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Reactivity data

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Product details

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The NOTCH3 protein a significant player in the NOTCH signaling pathway is a transmembrane receptor involved in cell differentiation. Sometimes referred to as 'Notch3' this protein has a molecular weight of approximately 244 kDa. NOTCH3 is widely expressed in vascular smooth muscle cells and tissues such as arteries and lungs. The protein also plays an important role in the communication between adjacent cells because of its location on the cell surface.
Biological function summary

NOTCH3 influences various cellular processes including proliferation apoptosis and differentiation. NOTCH3 does not function alone it is part of a regulatory complex that mediates cellular context and fate decisions. The receptor is cleaved upon ligand binding to facilitate intracellular signaling. This signaling is critical in the development and maintenance of vascular integrity which highlights the protein's importance in both physiological and pathological contexts.

Pathways

NOTCH3 plays a role in the NOTCH signaling pathway essential for cell fate determination and the smooth muscle cell differentiation pathway. Ligands such as Jagged and Delta-like proteins trigger NOTCH3 activation. Upon activation the NOTCH3 intracellular domain translocates to the nucleus where it interacts with transcriptional regulators like RBPJ. This interaction modulates the expression of target genes influencing cell differentiation and proliferation.

NOTCH3 is implicated in Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) a genetic vascular disorder. Mutations in the NOTCH3 gene result in the accumulation of the NOTCH3 protein within arterial walls leading to the characteristic symptoms of CADASIL. Additionally the protein has been linked to pulmonary arterial hypertension through shared pathways with proteins such as BMP2 and SMADs indicating its broader impact on vascular pathophysiology.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination (PubMed : 15350543, PubMed : 14714274). 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 (By similarity).
See full target information NOTC3_HUMAN
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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