Recombinant Anti-Nicastrin antibody [EPR2575Y] (ab68145)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2575Y] to Nicastrin
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Nicastrin antibody [EPR2575Y]
See all Nicastrin primary antibodies -
Description
Rabbit monoclonal [EPR2575Y] to Nicastrin -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Nicastrin aa 650-750 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HEK293T, HeLa, Jurkat and HL-60 cell lysates; C6 cell lysate; Neuro-2a cell lysate; U-87 MG and SH-SY5Y cell lysates.
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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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2575Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68145 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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WB |
1/1000 - 1/2000. Detects a band of approximately 100-120, 130-160 kDa (predicted molecular weight: 78 kDa).
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Notes |
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WB
1/1000 - 1/2000. Detects a band of approximately 100-120, 130-160 kDa (predicted molecular weight: 78 kDa). |
Target
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Function
Essential subunit of the gamma-secretase complex, an endoprotease complex that catalyzes the intramembrane cleavage of integral membrane proteins such as Notch receptors and APP (beta-amyloid precursor protein). It probably represents a stabilizing cofactor required for the assembly of the gamma-secretase complex. -
Tissue specificity
Widely expressed. -
Sequence similarities
Belongs to the nicastrin family. -
Cellular localization
Membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 23385 Human
- Entrez Gene: 59287 Mouse
- Entrez Gene: 289231 Rat
- Omim: 605254 Human
- SwissProt: Q92542 Human
- SwissProt: P57716 Mouse
- SwissProt: Q8CGU6 Rat
- Unigene: 517249 Human
see all -
Alternative names
- Anterior pharynx defective 2 antibody
- APH 2 antibody
- APH2 antibody
see all
Images
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All lanes : Anti-Nicastrin antibody [EPR2575Y] (ab68145) at 1/1000 dilution
Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 2 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 4 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 78 kDa
Observed band size: 100-140 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: Lane 1, 4: 180 seconds; Lane 2, 3: 15 seconds.
The bands observed is consistency with literature (PMID: 15322084).
The lysates were freshly made and used for Western Blotting immediately to minimize protein degradation. -
All lanes : Anti-Nicastrin antibody [EPR2575Y] (ab68145) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : NCSTN knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : HL-60 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 78 kDa
Observed band size: 100-125 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab68145 observed at 100-125 kDa. Red - loading control ab8245 observed at 36 kDa.
ab68145 Anti-Nicastrin antibody [EPR2575Y] was shown to specifically react with Nicastrin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266602 (knockout cell lysate ab258061) was used. Wild-type and Nicastrin knockout samples were subjected to SDS-PAGE. ab68145 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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.
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All lanes : Anti-Nicastrin antibody [EPR2575Y] (ab68145) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : HL-60 cell lysate
Lane 3 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP conjugated Goat anti-Rabbit at 1/2000 dilution
Predicted band size: 78 kDa
Additional bands at: 100-120 kDa (possible isoform), 130-160 kDa (possible isoform)
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab68145 has been referenced in 1 publication.
- Hsu YC et al. De-Glycyrrhizinated Licorice Extract Attenuates High Glucose-Stimulated Renal Tubular Epithelial-Mesenchymal Transition via Suppressing the Notch2 Signaling Pathway. Cells 9:N/A (2020). PubMed: 31948095