ATOX1 Recombinant Rabbit Monoclonal Antibody [PSH08-50]
Catalog# HA722996
ATOX1 Recombinant Rabbit Monoclonal Antibody [PSH08-50]
-
WB
-
IF-Cell
-
IHC-P
-
FC
-
Human
-
unconjugated
Overview
Product Name
ATOX1 Recombinant Rabbit Monoclonal Antibody [PSH08-50]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human ATOX1 aa 1-68.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, FC
Target Molecular Weight
Predicted band size: 7 kDa
Positive Control
HeLa cell lysate, 293T cell lysate, MCF7 cell lysate, NCI-H1299 cell lysate, 293T, human liver cancer tissue.
Conjugation
unconjugated
Clone Number
PSH08-50
Reactivity Data
Verified (internally validated)
Reported in literature (not internally validated)
Predicted reactive (based on sequence homology)
Not recommended (failed internal validation)
| WB | IHC-P | FC | IF-Cell | |
|---|---|---|---|---|
| human |
|
|
|
|
| mouse |
|
|
|
|
| rat |
|
|
|
|
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:2,000
-
IF-Cell
-
1:100
-
IHC-P
-
1:200
-
FC
-
1:1,000
Target
Function
ATOX1 is a copper metallochaperone protein that is encoded by the ATOX1 gene in humans. In mammals, ATOX1 plays a key role in copper homeostasis as it delivers copper from the cytosol to transporters ATP7A and ATP7B. Homologous proteins are found in a wide variety of eukaryotes, including Saccharomyces cerevisiae as ATX1, and all contain a conserved metal binding domain. ATOX1 is an abbreviation of the full name Antioxidant Protein 1. The nomenclature stems from initial characterization that showed that ATOX1 protected cells from reactive oxygen species. Since then, the primary role of ATOX1 has been established as a copper metallochaperone protein found in the cytoplasm of eukaryotes. A metallochaperone is an important protein that has metal trafficking and sequestration roles. As a metal sequestration protein, ATOX1 is capable of binding free metals in vivo, in order to protect cells from generation of reactive oxygen species and mismetallation of metalloproteins. As a metal trafficking protein, ATOX1 is responsible for shuttling copper from the cytosol to ATPase transporters ATP7A and ATP7B that move copper to the trans-Golgi network or secretory vesicles. In Saccharomyces cerevisiae, Atx1 delivers Cu(I) to a homologous transporter, Ccc2. The delivery of copper to ATPase transporters is vital for the subsequent insertion of copper into ceruloplasmin, a ferroxidase required for iron metabolism, within the golgi apparatus. In addition to the metallochaperone function, recent reports have characterized ATOX1 as a cyclin D1 transcription factor.
Background References
1. Chen L et al. APEX2-based Proximity Labeling of Atox1 Identifies CRIP2 as a Nuclear Copper-binding Protein that Regulates Autophagy Activation. Angew Chem Int Ed Engl. 2021 Nov
2. Zhao P et al. Atox1 protects hippocampal neurons after traumatic brain injury via DJ-1 mediated anti-oxidative stress and mitophagy. Redox Biol. 2024 Jun
Subcellular Location
Cytosol.
UNIPROT
Synonyms
ATOX1 antibody
ATOX1_HUMAN antibody
ATX1 antibody
ATX1 antioxidant protein 1 homolog (yeast) antibody
ATX1 antioxidant protein 1 homolog antibody
Copper transport protein antibody
Copper transport protein ATOX1 antibody
HAH1 antibody
Metal transport protein antibody
Metal transport protein ATX1 antibody
ExpandImages
-
Western blot analysis of ATOX1 on different lysates with Rabbit anti-ATOX1 antibody (HA722996) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: MCF7 cell lysate (20 µg/Lane)
Lane 4: NCI-H1299 cell lysate (10 µg/Lane)
Predicted band size: 7 kDa
Observed band size: 7 kDa
Exposure time: 11 seconds; ECL: K1801;
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 (HA722996) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of 293T cells labeling ATOX1 with Rabbit anti-ATOX1 antibody (HA722996) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATOX1 antibody (HA722996) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-ATOX1 antibody (HA722996) at 1/200 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 (HA722996) at 1/200 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 293T cells labeling ATOX1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA722996, 1/1,000) (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 iFluor™ 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"