ACY1 Mouse Monoclonal Antibody [15G2]
Catalog# EM1901-86
ACY1 Mouse Monoclonal Antibody [15G2]
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WB
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
ACY1 Mouse Monoclonal Antibody [15G2]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Recombinant protein within Human ACY1 aa 55-256 / 408.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC
Molecular Weight
Predicted band size: 46 kDa
Positive Control
K-562 cell lysate, HepG2 cell lysate, PC-3M cell lysate, human liver tissue lysate, human kidney tissue lysate, mouse liver tissue lysate, mouse kidney tissue lysate, rat liver tissue lysate, rat kidney tissue lysate, human kidney tissue, mouse kidney tissue, rat kidney tissue, SHSY5Y.
Conjugation
unconjugated
Clone Number
15G2
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG2a
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:1,000
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IHC-P
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1:2,000
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FC
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1:50-1:100
Target
Function
Aminoacylase is involved in the regulation of the urea cycle. N-acetyl-L-glutamate is an allosteric activator of carbamoyl phosphate synthetase, a crucial enzyme that commits NH4+ molecules to the urea cycle. The urea cycle gets rid of excess ammonia (NH4+) in the body, a process that must be up-regulated during times of increased protein catabolism, as amino acid breakdown produces large amounts of NH4+. When amino acid catabolism increases, N-Acetylglutamate synthase is up-regulated, producing more N-acetyl-L-glutamate, which up-regulates carbamoyl phosphate synthetase and allows it to dispose of the excess NH4+ from catabolism.Aminoacylase is up-regulated during times of nutrient deficit or starvation, causing N-acetyl-L-glutamate breakdown, which down-regulates carbamoyl phosphate synthetase and the rest of the urea cycle. This response is evolutionarily advantageous, since a nutrient deficit means there isn't as much NH4+ that needs to be disposed of and since the body wants to salvage as many amino acids as it can.
Background References
1. Lindner H.A.et.al.Essential roles of zinc ligation and enzyme dimerization for catalysis in the aminoacylase-1/M20 family.J. Biol. Chem. 278:44496-44504(2003).
Sequence Similarity
Belongs to the peptidase M20A family.
Tissue Specificity
Expression is highest in kidney, strong in brain and weaker in placenta and spleen.
Subcellular Location
Cytoplasm.
Synonyms
ACY 1 antibody
ACY-1 antibody
Acy1 antibody
ACY1_HUMAN antibody
ACY1D antibody
ACYLASE antibody
Acylase I antibody
Aminoacylase 1 antibody
Aminoacylase-1 antibody
EC 3.5.1.14 antibody
ExpandImages
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Western blot analysis of ACY1 on different lysates with Mouse anti-ACY1 antibody (EM1901-86) at 1/1,000 dilution.
Lane 1: K-562 cell lysate (20 µg/Lane)
Lane 2: HepG2 cell lysate (20 µg/Lane)
Lane 3: PC-3M cell lysate (20 µg/Lane)
Lane 4: Human liver tissue lysate (40 µg/Lane)
Lane 5: Human kidney tissue lysate (40 µg/Lane)
Lane 6: Mouse liver tissue lysate (40 µg/Lane)
Lane 7: Mouse kidney tissue lysate (40 µg/Lane)
Lane 8: Rat liver tissue lysate (40 µg/Lane)
Lane 9: Rat kidney tissue lysate (40 µg/Lane)
Predicted band size: 46 kDa
Observed band size: 42 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-86) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of ACY1 on different lysates with Mouse anti-ACY1 antibody (EM1901-86) at 1/1,000 dilution.
Lane 1: A549-si NT cell lysate
Lane 2: A549-si ACY1 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 3 minutes; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-86) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-ACY1 antibody (EM1901-86) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-86) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-ACY1 antibody (EM1901-86) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-86) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-ACY1 antibody (EM1901-86) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-86) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of ACY1 was done on SHSY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-86, 1/100) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa FluorTM488 Goat anti-Mouse IgG IgG Secondary antibody at 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"