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rabbit anti-BACE (CT) polyclonal antibody 7999

$445.00

Antibody summary

  • Rabbit polyclonal to BACE (CT)
  • Suitable for: ELISA,WB,ICC,IHC-P,IF
  • Isotype: IgG
  • 100 µg
SKU: 7999parent Category: Tags: , ,
Weight1 lbs
Dimensions9 × 5 × 2 in
host

rabbit

isotype

IgG

clonality

polyclonal

concentration

1 mg/mL

applications

ICC/IF, WB

reactivity

BACE (CT)

available sizes

100 µg

rabbit anti-BACE (CT) polyclonal antibody 7999

antibody
Tested applications
WB,IHC,IHC,ICC/IF,ELISA
Recommended dilutions
Immunoblotting: use at 1:500-1:1,000 dilution.

Positive control: Human brain tissue lysate.
Immunogen
Peptide corresponding to aa 485-501 of human BACE. This sequence differs from those of mouse and rat by one amino acid.
Size and concentration
100µg and lot specific
Form
liquid
Storage Instructions
This antibody is stable for at least one (1) year at -20°C. Avoid multiple freeze-thaw cycles.
Storage buffer
PBS, pH 7.4.
Purity
immunogen affinty purifcation
Clonality
polyclonal
Isotype
IgG
Compatible secondaries
goat anti-rabbit IgG, H&L chain specific, peroxidase conjugated, conjugated polyclonal antibody 9512
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody 2079
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody 7863
goat anti-rabbit IgG, H&L chain specific, Cross Absorbed polyclonal antibody 2371
goat anti-rabbit IgG, H&L chain specific, biotin conjugated polyclonal antibody, crossabsorbed 1715
goat anti-rabbit IgG, H&L chain specific, FITC conjugated polyclonal antibody, crossabsorbed 1720
Isotype control
Rabbit polyclonal - Isotype Control
target relevance
Protein names
Beta-secretase 1 (EC 3.4.23.46) (Aspartyl protease 2) (ASP2) (Asp 2) (Beta-site amyloid precursor protein cleaving enzyme 1) (Beta-site APP cleaving enzyme 1) (Memapsin-2) (Membrane-associated aspartic protease 2)
Gene names
BACE1,BACE1 BACE KIAA1149
Protein family
Peptidase A1 family
Mass
55764Da
Function
FUNCTION: Responsible for the proteolytic processing of the amyloid precursor protein (APP). Cleaves at the N-terminus of the A-beta peptide sequence, between residues 671 and 672 of APP, leads to the generation and extracellular release of beta-cleaved soluble APP, and a corresponding cell-associated C-terminal fragment which is later released by gamma-secretase (PubMed:10656250, PubMed:10677483, PubMed:20354142). Cleaves CHL1 (By similarity). {ECO:0000250|UniProtKB:P56818, ECO:0000269|PubMed:10656250, ECO:0000269|PubMed:10677483, ECO:0000269|PubMed:20354142}.
Catalytic activity
CATALYTIC ACTIVITY: Reaction=Broad endopeptidase specificity. Cleaves Glu-Val-Asn-Leu-|-Asp-Ala-Glu-Phe in the Swedish variant of Alzheimer's amyloid precursor protein.; EC=3.4.23.46; Evidence={ECO:0000269|PubMed:10677483};
Subellular location
SUBCELLULAR LOCATION: Cell membrane {ECO:0000269|PubMed:11466313}; Single-pass type I membrane protein {ECO:0000305|PubMed:11466313}. Golgi apparatus, trans-Golgi network {ECO:0000269|PubMed:11466313, ECO:0000269|PubMed:15615712, ECO:0000269|PubMed:15886016, ECO:0000269|PubMed:17425515, ECO:0000269|PubMed:20354142, ECO:0000269|PubMed:23109336}. Endoplasmic reticulum {ECO:0000269|PubMed:11466313, ECO:0000269|PubMed:17425515}. Endosome {ECO:0000269|PubMed:11466313, ECO:0000269|PubMed:15886016}. Cell surface {ECO:0000269|PubMed:11466313, ECO:0000269|PubMed:15886016, ECO:0000269|PubMed:17425515, ECO:0000269|PubMed:23109336}. Cytoplasmic vesicle membrane {ECO:0000269|PubMed:11466313, ECO:0000269|PubMed:15886016}; Single-pass type I membrane protein {ECO:0000305|PubMed:11466313}. Membrane raft {ECO:0000250|UniProtKB:P56818}. Lysosome {ECO:0000269|PubMed:16033761, ECO:0000269|PubMed:23109336, ECO:0000269|PubMed:27084579, ECO:0000269|PubMed:27302062}. Late endosome {ECO:0000269|PubMed:16033761, ECO:0000269|PubMed:23109336, ECO:0000269|PubMed:27084579, ECO:0000269|PubMed:27302062}. Early endosome {ECO:0000269|PubMed:15615712, ECO:0000269|PubMed:15886016, ECO:0000269|PubMed:23109336, ECO:0000269|PubMed:27084579, ECO:0000269|PubMed:27302062}. Recycling endosome {ECO:0000269|PubMed:15886016, ECO:0000269|PubMed:27084579, ECO:0000269|PubMed:27302062}. Cell projection, axon {ECO:0000250|UniProtKB:P56818}. Cell projection, dendrite {ECO:0000250|UniProtKB:P56818}. Note=Predominantly localized to the later Golgi/trans-Golgi network (TGN) and minimally detectable in the early Golgi compartments. A small portion is also found in the endoplasmic reticulum, endosomes and on the cell surface (PubMed:17425515, PubMed:11466313). Colocalization with APP in early endosomes is due to addition of bisecting N-acetylglucosamine wich blocks targeting to late endosomes and lysosomes (By similarity). Retrogradly transported from endosomal compartments to the trans-Golgi network in a phosphorylation- and GGA1- dependent manner (PubMed:15886016). {ECO:0000250|UniProtKB:P56818, ECO:0000269|PubMed:11466313, ECO:0000269|PubMed:15886016, ECO:0000269|PubMed:17425515}.
Tissues
TISSUE SPECIFICITY: Expressed at high levels in the brain and pancreas. In the brain, expression is highest in the substantia nigra, locus coruleus and medulla oblongata. {ECO:0000269|PubMed:10677483, ECO:0000269|PubMed:11083922}.
Structure
SUBUNIT: Monomer. Interacts (via DXXLL motif) with GGA1, GGA2 and GGA3 (via their VHS domain); the interaction highly increases when BACE1 is phosphorylated at Ser-498 (PubMed:14567678, PubMed:15886016). Interacts with RTN1; RTN2; RTN3 and RTN4; the interaction leads to inhibition of amyloid precursor protein processing (PubMed:15286784, PubMed:16965550, PubMed:16979658). Interacts with SNX6 (PubMed:20354142). Interacts with PCSK9 (PubMed:18660751). Interacts with NAT8 and NAT8B (PubMed:19011241). Interacts with BIN1 (PubMed:27179792). Interacts (via extracellular domain) with ADAM10 (via extracellular domain) (By similarity). Interacts with SORL1; this interaction may affect binding with APP and hence reduce APP cleavage (PubMed:16407538). Interacts with NRDC AND NRG1 (By similarity). {ECO:0000250|UniProtKB:P56818, ECO:0000269|PubMed:14567678, ECO:0000269|PubMed:15286784, ECO:0000269|PubMed:15886016, ECO:0000269|PubMed:16407538, ECO:0000269|PubMed:16965550, ECO:0000269|PubMed:16979658, ECO:0000269|PubMed:18660751, ECO:0000269|PubMed:19011241, ECO:0000269|PubMed:20354142, ECO:0000269|PubMed:27179792}.
Post-translational modification
PTM: N-Glycosylated (PubMed:11083922, PubMed:17425515). Addition of a bisecting N-acetylglucosamine by MGAT3 blocks lysosomal targeting, further degradation and is required for maintaining stability under stress conditions (By similarity). {ECO:0000250|UniProtKB:P56818, ECO:0000269|PubMed:11083922, ECO:0000269|PubMed:17425515}.; PTM: Acetylated in the endoplasmic reticulum at Lys-126, Lys-275, Lys-279, Lys-285, Lys-299, Lys-300 and Lys-307. Acetylation by NAT8 and NAT8B is transient and deacetylation probably occurs in the Golgi. Acetylation regulates the maturation, the transport to the plasma membrane, the stability and the expression of the protein. {ECO:0000269|PubMed:17425515, ECO:0000269|PubMed:19011241, ECO:0000269|PubMed:20826464}.; PTM: Palmitoylation mediates lipid raft localization. {ECO:0000250|UniProtKB:P56818}.; PTM: Ubiquitinated at Lys-501, ubiquitination leads to lysosomal degradation (PubMed:27302062, PubMed:16033761, PubMed:20484053, PubMed:23109336). Monoubiquitinated and 'Lys-63'-linked polyubitinated (PubMed:20484053). Deubiquitnated by USP8; inhibits lysosomal degradation (PubMed:27302062). {ECO:0000269|PubMed:16033761, ECO:0000269|PubMed:20484053, ECO:0000269|PubMed:23109336, ECO:0000269|PubMed:27302062}.; PTM: Phosphorylation at Ser-498 is required for interaction with GGA1 and retrograded transport from endosomal compartments to the trans-Golgi network. Non-phosphorylated BACE1 enters a direct recycling route to the cell surface. {ECO:0000269|PubMed:15886016}.
Domain
TOPO_DOM 2
Target Relevance information above includes information from UniProt accession : P56817
The UniProt Consortium

Data

benchmark-antibodies_anti-bace_ct_antibody_7999_1.jpg
Western Blot Validation of BACE
Loading: 15 µg of lysates per lane. Antibodies: BACE (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane A-C: human brain tissue lysate in the absence (A) or presence (B) of blocking peptide and mouse 3T3/NIH cell lysate (C).
benchmark-antibodies_anti-bace_ct_antibody_7999_2.gif
Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines
Loading: 15 µg of lysates per lane. Antibodies: BACE 7999 (1 µg/mL), BACE 32-238 (1 µg/mL), beta-actin (1 µg/mL), and GAPDH (0.02 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
benchmark-antibodies_anti-bace_ct_antibody_7999_3.gif
Immunohistochemistry Validation of BACE in Mouse Brain
Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-BACE antibody (7999) at 2.5 µg/mL. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-bace_ct_antibody_7999_4.gif
Immunofluorescence Validation of BACE in 3T3/NIH Cells
Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse 3T3/NIH cells labeling BACE with 7999 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (green) and DAPI staining (blue). Image showing both membrane and cytosol staining on 3T3/NIH cells.
benchmark-antibodies_anti-bace_ct_antibody_7999_5.jpg
Immunocytochemistry Validation of BACE in 3T3/NIH Cells
Immunocytochemical analysis of 3T3/NIH cells using anti-BACE antibody (7999) at 10 µg/mL. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
benchmark-antibodies_anti-bace_ct_antibody_7999_6.gif
KO and Overexpression Validation of BACE in Human and Mouse Brain and 293 Cells. (Singer et al., 2005)
Western blot analysis of the BACE1 (7999) antibody?s ability to recognize human and murine BACE1. The BACE1 antibody recognized both the mouse and human forms of BACE1. Lanes 1?4 are frontal cortex homogenates from human and mouse brains. Lane 1 is from a neurologically unimpaired aged human control case, lane 2 from a BACE1-deficient mouse, lane 3 from a nontransgenic mouse and lane 4 from hBACE1 transgenic mouse. Lanes 5?7 are lysates from HEK293T cells transfected with a plasmid vector expressing eGFP, mBACE1 and hBACE1, respectively.
benchmark-antibodies_anti-bace_ct_antibody_7999_7.gif
KD Validation of BACE in Mouse Brain (Singer et al., 2005)
Characterization of the effects of lenti-siBACE1-6 expression in the brains of APP transgenic mice. (a?d) Anti-eGFP immunoreactivity in the hippocampus (the injection site) shows comparable and consistent expression of lenti-siRNA constructs in the dentate gyrus (dg) and stratus polymorphus (sp). (e) Anti-BACE1 immunoreactivity in the hippocampus of nontransgenic mice treated with lenti-siGlut4. (f) Reduced BACE1 immunostaining in the hippocampus of nontransgenic mice treated with lenti-siBACE1-6 vector. (g) Intense BACE1 immunoreactivity in the hippocampus of APP transgenic mice treated with lenti-siGlut4. (h) Reduced BACE1 expression in APP transgenic mice treated with lenti-siBACE1-6 vector. (i,j) Anti-BACE1 reacted with pyramidal cell bodies in the neocortex, which was not injected,
benchmark-antibodies_anti-bace_ct_antibody_7999_8.gif
KD Validation of BACE in Mouse Brain (Singer et al., 2005)
Immunolabeling patterns of BACE1 expression and the lenti-siRNA distribution. Sections from APP transgenic mice treated with the eGFPtagged lenti siRNA vectors (green) were co-immunolabeled with an antibody against BACE1 (red) and imaged with the LSCM. All sections are from the hippocampus of treated mice. (a?c) Lenti-siBACE1-6?treated mice. Areas within the hippocampus expressing the eGFP tagged vector have reduced BACE1 immunolabeling. (d?f) Mice treated with the eGFP-tagged control lenti-siGlut4 show unchanged expression of BACE1 in the hippocampus. (g?i) Mice treated with a saline vehicle show unchanged expression of BACE1 in the hippocampus.
benchmark-antibodies_anti-bace_ct_antibody_7999_9.gif
KO Validation of BACE in MEF Cells (Jo et al., 2010)
Wildtype and BACE ?/? MEFs were exposed to HNE (15_M) for 2 h. BACE1 levels were examined by Western blot with anti-BACE antibodies (7999).

Publications

Published literature highly relevant to the biological target of this product and referencing this antibody or clone are retrieved from PubMed database provided by The United States National Library of Medicine at the National Institutes of Health.




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Protocols

relevant to this product
Western blot
IHC
ICC

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