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Carboxypeptidase E (CPE) (AA 49-200) antibody

Details for Product No. ABIN968159
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Antigen
Synonyms CPE, CPH, Cph-1, Cph1, R74677, fat, CARBE, Cph, si:ch211-198g9.2, zgc:85981
Epitope
AA 49-200
(8), (6), (6), (2), (1), (1), (1), (1), (1), (1)
Reactivity
Human
(52), (31), (30), (15), (13), (13), (12), (2), (2), (1), (1), (1)
Host
Mouse
(51), (2), (1)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(3), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Immunofluorescence (IF), Immunohistochemistry (IHC)
(45), (24), (13), (12), (10), (10), (10), (6), (5), (1)
Pubmed 4 references available
Quantity 50 µg
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Catalog No. ABIN968159
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Immunogen Human Carboxypeptidase E
Clone G52C7-1
Isotype IgG1
Cross-Reactivity Rat (Rattus), Mouse (Murine)
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Alternative Name Carboxypeptidase E
Background Carboxypeptidase E (CPE), also known as carboxypeptidase H and enkephalin convertase, is found as both a membrane-bound and a soluble glycoprotein in neuroendocrine tissues and adrenal-gland chromaffin granules. The C-terminus forms an amphiphilic alpha-helix, suggesting that this region is responsible for the membrane-bound form. Evidence suggests the active form of CPE is located in the secretory vesicles. CPE appears to have several functions. It is an exopeptidase that cleaves neuropeptides with C-terminal basic amino acids, producing an active form of the peptide. It has also been proposed that membrane-bound CPE is a sorting receptor for regulated secretory pathway (RSP) proteins in the TGN pituitary Golgi and secretory granule membranes. RSP proteins primarily consist of hormones and neuropeptides. Mice that carry a mutation in the CPE gene Cpe[fat] display endocrine disorders such as obesity, infertility, and hyperproinsulinemia. Furthermore, the same endocrine disorders are observed in Cpe[fat] mice where the CPE gene has been effaced by antisense RNA.
Molecular Weight 50 kDa
Research Area Enzymes, Metabolism
Application Notes Methanol Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS.
Triton-X 100 Procedure for a 96 well plate: Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS.
Comment

Related Products: ABIN968545

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage -20 °C
Supplier Images
anti-Carboxypeptidase E (CPE) (AA 49-200) antibody Western blot analysis of Carboxypeptidase E on rat brain lysate (First Panel). Lane 1: 1:2500, lane 2: 1:5000, lane 3: 1:10000 dilution of Carboxypeptidase E,
anti-Carboxypeptidase E (CPE) (AA 49-200) antibody (2) Immunofluorescent staining of SH-SY5Y cells (Second Panel). Cells were seeded in a collagen coated 384 well imaging plate at ~ 8,000 cells per well. After overnight incubation, cells were stained using the Triton X100 fix/perm protocol and the anti-Carboxypeptidase E antibody. The second step reagent was Alexa Fluor® 488 goat anti mouse Ig (Invitrogen). The image was taken on a Pathway 855 or 435 imager using a 20x objective. This antibody also stained SK-N-SH and C6 cells using both the Triton X100 and methanol fix/perm protocols.
Product cited in: Manser, Fernandez, Loo et al.: "Human carboxypeptidase E. Isolation and characterization of the cDNA, sequence conservation, expression and processing in vitro." in: The Biochemical journal, Vol. 267, Issue 2, pp. 517-25, 1990 (PubMed).

Varlamov, Fricker: "The C-terminal region of carboxypeptidase E involved in membrane binding is distinct from the region involved with intracellular routing." in: The Journal of biological chemistry, Vol. 271, Issue 11, pp. 6077-83, 1996 (PubMed).

Cool, Normant, Shen et al.: "Carboxypeptidase E is a regulated secretory pathway sorting receptor: genetic obliteration leads to endocrine disorders in Cpe(fat) mice." in: Cell, Vol. 88, Issue 1, pp. 73-83, 1997 (PubMed).

Shen, Loh: "Intracellular misrouting and abnormal secretion of adrenocorticotropin and growth hormone in cpefat mice associated with a carboxypeptidase E mutation." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 94, Issue 10, pp. 5314-9, 1997 (PubMed).

Validation Images
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