PAH Recombinant Rabbit Monoclonal Antibody [JE64-47]
Catalog# HA721122
PAH Recombinant Rabbit Monoclonal Antibody [JE64-47]
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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
PAH Recombinant Rabbit Monoclonal Antibody [JE64-47]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human PAH aa 50-100/452.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC
Molecular Weight
Predicted band size: 52 kDa
Positive Control
Rat liver tissue lysates, rat kidney tissue, mouse kidney tissue, HepG2.
Conjugation
unconjugated
Clone Number
JE64-47
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IHC-P
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1:100-1:400
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FC
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1:500-1:1,000
Target
Function
Phenylalanine hydroxylase (PAH) (EC 1.14.16.1) is an enzyme that catalyzes the hydroxylation of the aromatic side-chain of phenylalanine to generate tyrosine. PAH is one of three members of the biopterin-dependent aromatic amino acid hydroxylases, a class of monooxygenase that uses tetrahydrobiopterin (BH4, a pteridine cofactor) and a non-heme iron for catalysis. During the reaction, molecular oxygen is heterolytically cleaved with sequential incorporation of one oxygen atom into BH4 and phenylalanine substrate. In humans, mutations in its encoding gene, PAH, can lead to the metabolic disorder phenylketonuria. Deficiency in PAH activity due to mutations in PAH causes hyperphenylalaninemia (HPA). Since phenylketonuria can cause irreversible damage, it is imperative that deficiencies in the phenylalanine hydroxylase are determined early on in development. Originally, this was done using a bacterial inhibition assay known as the Guthrie Test. Now, PKU is part of newborn screening in many countries, and elevated phenylalanine levels are identified shortly after birth by measurement with tandem mass spectrometry. Placing the individual on a low phenylalanine, high tyrosine diet can help prevent any long-term damage to their development.
Background References
1. Mitchell SC. et. al. Phenylalanine 4-monooxygenase: the "sulfoxidation polymorphism". Xenobiotica. 2020 Jan
2. Steventon GB. et. al. Phenylalanine 4-monooxygenase and the role of endobiotic metabolism enzymes in xenobiotic biotransformation. Expert Opin Drug Metab Toxicol. 2009 Oct
Subcellular Location
Cytosol.
Synonyms
PAH antibody
PH antibody
PH4H_HUMAN antibody
Phe 4 monooxygenase antibody
Phe-4-monooxygenase antibody
Phenylalanine 4 hydroxylase antibody
Phenylalanine hydroxylase antibody
Phenylalanine-4-hydroxylase antibody
PKU antibody
PKU1 antibody
Images
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Western blot analysis of PAH on Rat liver tissue lysate with Rabbit anti-PAH antibody (HA721122) at 1/1,000 dilution.
Lysates/proteins at 10 µg/Lane.
Exposure time: 10 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA721122, 1/1,000 in 5% NFDM/TBST, 2 hours at room temperature
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 52 kDa
Observed band size: 52 kDa -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-PAH antibody (HA721122) at 1/100 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 (HA721122) at 1/100 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 Rabbit anti-PAH antibody (HA721122) at 1/400 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 (HA721122) at 1/400 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 HepG2 cells labeling PAH.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721122, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a Alexa Fluor® 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. 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"