製品: GPX4 Antibody
カタログ: DF6701
タンパク質の説明: Rabbit polyclonal antibody to GPX4
アプリケーション: WB IHC IF/ICC
反応性: Human, Mouse, Rat
予測: Pig, Bovine, Chicken
分子量: 22kDa; 22kD(Calculated).
ユニプロット: P36969
RRID: AB_2838663

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製品説明

ソース:
Rabbit
アプリケーション:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat
予測:
Pig(90%), Bovine(100%), Chicken(80%)
クローナリティ:
Polyclonal
特異性:
GPX4 Antibody detects endogenous levels of total GPX4.
RRID:
AB_2838663
引用形式: Affinity Biosciences Cat# DF6701, RRID:AB_2838663.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ/展開

Glutathione peroxidase 4; GPX 4; GPX-4; GPX4; GPX4_HUMAN; GSHPx-4; MCSP; mitochondrial; PHGPx; Phospholipid hydroperoxidase; Phospholipid hydroperoxide glutathione peroxidase; Phospholipid hydroperoxide glutathione peroxidase mitochondrial; snGPx; snPHGPx; Sperm nucleus glutathione peroxidase;

免疫原

免疫原:

A synthesized peptide derived from human GPX4, corresponding to a region within C-terminal amino acids.

Uniprot:
遺伝子(ID):
発現特異性:
P36969 GPX4_HUMAN:

Present primarily in testis.

タンパク質の説明:
Glutathione peroxidase catalyzes the reduction of hydrogen peroxide, organic hydroperoxide, and lipid peroxides by reduced glutathione and functions in the protection of cells against oxidative damage. Human plasma glutathione peroxidase has been shown to be a selenium-containing enzyme and the UGA codon is translated into a selenocysteine. Through alternative splicing and transcription initiation, rat produces proteins that localize to the nucleus, mitochondrion, and cytoplasm. In humans, experimental evidence for alternative splicing exists; alternative transcription initiation and the cleavage sites of the mitochondrial and nuclear transit peptides need to be experimentally verified. [provided by RefSeq, Jul 2008]
タンパク質配列:
MSLGRLCRLLKPALLCGALAAPGLAGTMCASRDDWRCARSMHEFSAKDIDGHMVNLDKYRGFVCIVTNVASQUGKTEVNYTQLVDLHARYAECGLRILAFPCNQFGKQEPGSNEEIKEFAAGYNVKFDMFSKICVNGDDAHPLWKWMKIQPKGKGILGNAIKWNFTKFLIDKNGCVVKRYGPMEEPLVIEKDLPHYF

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Bovine
100
Pig
90
Chicken
80
Horse
0
Sheep
0
Dog
0
Xenopus
0
Zebrafish
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P36969 基板として

Site PTM Type Enzyme
S40 Phosphorylation
K47 Acetylation
K107 Ubiquitination
K162 Ubiquitination
K167 Ubiquitination

研究背景

機能:

Essential antioxidant peroxidase that directly reduces phospholipid hydroperoxide even if they are incorporated in membranes and lipoproteins (By similarity). Can also reduce fatty acid hydroperoxide, cholesterol hydroperoxide and thymine hydroperoxide (By similarity). Plays a key role in protecting cells from oxidative damage by preventing membrane lipid peroxidation (By similarity). Required to prevent cells from ferroptosis, a non-apoptotic cell death resulting from an iron-dependent accumulation of lipid reactive oxygen species. The presence of selenocysteine (Sec) versus Cys at the active site is essential for life: it provides resistance to overoxidation and prevents cells against ferroptosis (By similarity). The presence of Sec at the active site is also essential for the survival of a specific type of parvalbumin-positive interneurons, thereby preventing against fatal epileptic seizures (By similarity). May be required to protect cells from the toxicity of ingested lipid hydroperoxides (By similarity). Required for normal sperm development and male fertility (By similarity). Essential for maturation and survival of photoreceptor cells (By similarity). Plays a role in a primary T-cell response to viral and parasitic infection by protecting T-cells from ferroptosis and by supporting T-cell expansion (By similarity).

細胞の位置付け:

Mitochondrion.

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Present primarily in testis.

サブユニット構造:

Monomer. Has a tendency to form higher mass oligomers.

タンパク質ファミリー:

Belongs to the glutathione peroxidase family.

研究領域

· Cellular Processes > Cell growth and death > Ferroptosis.   (View pathway)

· Metabolism > Metabolism of other amino acids > Glutathione metabolism.

参考文献

1). Arsenic retention in erythrocytes and excessive erythrophagocytosis is related to low selenium status by impaired redox homeostasis. Redox Biology, 2022 (PubMed: 35500533) [IF=11.4]

2). BSA-stabilized selenium nanoparticles ameliorate intracerebral hemorrhage's-like pathology by inhibiting ferroptosis-mediated neurotoxicology via Nrf2/GPX4 axis activation. Redox biology, 2024 (PubMed: 39032396) [IF=11.4]

3). ZnO NPs induce miR-342-5p mediated ferroptosis of spermatocytes through the NF-κB pathway in mice. JOURNAL OF NANOBIOTECHNOLOGY, 2024 [IF=10.8]

Application: WB    Species: Mouse    Sample: GC-2 cells

Fig. 6 ZnO NPs induce the ferroptosis of GC-2 cells. (A) Intracellular chelatable iron in GC-2 cells treated with or without ZnO NPs stained with PGSK (green). Statistical analysis of MFI of PGSK was shown. (B) Representative FACS data for lipid peroxidation level in GC-2 cells following ZnO NPs treatment using C11 BODIPY. Statistical analysis of MFI of the ratio of green/red was shown. (C) qRT-PCR analysis of ferroptosis-related gene expression in GC-2 cells after ZnO NPs treatment. (D) Western blot of ferroptosis-related protein levels in GC-2 cells treated with ZnO NPs. Statistical analysis of mean grey values ratios of the corresponding proteins/β-actin was shown, the same as below. (E) The cell viability of GC-2 cells following ZnO NPs treatment with or without Fer-1 (3.5 µM). (F) Intracellular chelatable iron in GC-2 cells following ZnO NPs treatment with or without Fer-1 stained with PGSK. Statistical analysis of MFI of PGSK was shown. (G) Representative FACS data of lipid peroxidation level in GC-2 cells following ZnO NPs treatment with or without Fer-1 stained with C11 BODIPY. Statistical analysis of MFI of the ratio of green/red was shown. (H and I) The levels of GSH and MDA in GC-2 cells following ZnO NPs treatment with or without Fer-1. (J) qRT-PCR analysis of ferroptosis-related gene expression in GC-2 cells following ZnO NPs treatment with or without Fer-1. (K) Western blot of ferroptosis-related protein levels in GC-2 cells following ZnO NPs treatment with or without Fer-1

4). Silibinin attenuates ferroptosis in acute kidney injury by targeting FTH1. Redox Biology, 2024 [IF=10.7]

5). Rationally designed catalytic nanoplatform for enhanced chemoimmunotherapy via deploying endogenous plus exogenous copper and remodeling tumor microenvironment. Journal of nanobiotechnology, 2024 (PubMed: 39252079) [IF=10.2]

6). Cell Membrane Camouflaged Metal Oxide–Black Phosphorus Biomimetic Nanocomplex Enhances Photo-chemo-dynamic Ferroptosis. ACS Applied Materials & Interfaces, 2022 (PubMed: 35658416) [IF=9.5]

7). Microbial metabolite deoxycholic acid-mediated ferroptosis exacerbates high-fat diet-induced colonic inflammation. Molecular metabolism, 2024 (PubMed: 38642891) [IF=8.1]

Application: IHC    Species: Mouse    Sample: colonic tissues

Figure 3 Deoxycholic acid enhanced lipopolysaccharide-induced ferroptosis in intestinal epithelial cells (A–H) IEC-6 cells and Caco-2 cells were incubated with or without 10 ng/mL LPS for 12 h, followed by 200 μM DCA for 2 h. They were divided into four group (n = 6 in each group): control group, DCA group, LPS group and LPS + DCA group. (A) The relative mRNA expression level of GPX4 in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (B) The relative mRNA expression level of ACSL4 in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (C) The protein level of GPX4, ACSL4 and β-actin in each group of IEC-6 cells and Caco-2 cells. (D) Representative transmission electron microscopy images (scale bars, 1 μm) of cell and mitochondria, and Fe2+/Hoechst immunofluorescence in each group of IEC-6 cells. Mitochondria was showed by white arrows in the pictures. The red fluorescence is indicative of the presence of ferrous ions. (E) The content of Fe2+ in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (F) The content of GSH in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (G) The content of ROS in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (H) The content of MDA in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (I-K) IEC-6 cells were incubated 2 μM ferrostatin-1 (Fer-1) for 16 h, and then incubated with or without 10 ng/mL LPS for 12 h, followed by 200 μM DCA for 2 h. They were divided into six group (n = 5 or 6 in each group): control group, LPS group, LPS + DCA group, Fer-1 group, LPS + Fer-1 group and LPS + DCA + Fer-1 group. (I) The relative mRNA expression level of GPX4, ACSL4 and DMT1 in each group of IEC-6 cells (n = 6 in each group). (J) The content of GSH in each group of IEC-6 cells (n = 5 in each group). (K) The content of MDA in each group of IEC-6 cells (n = 5 in each group). Data are represented as mean ± SEM. ns P > 0.05, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. ##P < 0.01, ####P < 0.0001. ∗ LPS group vs control group, LPS + DCA group vs LPS group. # LPS + Fer-1 group vs LPS group, LPS + DCA + Fer-1 group vs LPS + DCA group.

Application: WB    Species: Rat and Human    Sample: IEC-6 cells and Caco-2 cells

Figure 3 Deoxycholic acid enhanced lipopolysaccharide-induced ferroptosis in intestinal epithelial cells (A–H) IEC-6 cells and Caco-2 cells were incubated with or without 10 ng/mL LPS for 12 h, followed by 200 μM DCA for 2 h. They were divided into four group (n = 6 in each group): control group, DCA group, LPS group and LPS + DCA group. (A) The relative mRNA expression level of GPX4 in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (B) The relative mRNA expression level of ACSL4 in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (C) The protein level of GPX4, ACSL4 and β-actin in each group of IEC-6 cells and Caco-2 cells. (D) Representative transmission electron microscopy images (scale bars, 1 μm) of cell and mitochondria, and Fe2+/Hoechst immunofluorescence in each group of IEC-6 cells. Mitochondria was showed by white arrows in the pictures. The red fluorescence is indicative of the presence of ferrous ions. (E) The content of Fe2+ in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (F) The content of GSH in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (G) The content of ROS in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (H) The content of MDA in each group of IEC-6 cells and Caco-2 cells (n = 6 in each group). (I-K) IEC-6 cells were incubated 2 μM ferrostatin-1 (Fer-1) for 16 h, and then incubated with or without 10 ng/mL LPS for 12 h, followed by 200 μM DCA for 2 h. They were divided into six group (n = 5 or 6 in each group): control group, LPS group, LPS + DCA group, Fer-1 group, LPS + Fer-1 group and LPS + DCA + Fer-1 group. (I) The relative mRNA expression level of GPX4, ACSL4 and DMT1 in each group of IEC-6 cells (n = 6 in each group). (J) The content of GSH in each group of IEC-6 cells (n = 5 in each group). (K) The content of MDA in each group of IEC-6 cells (n = 5 in each group). Data are represented as mean ± SEM. ns P > 0.05, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001. ##P < 0.01, ####P < 0.0001. ∗ LPS group vs control group, LPS + DCA group vs LPS group. # LPS + Fer-1 group vs LPS group, LPS + DCA + Fer-1 group vs LPS + DCA group.

8). Vitamin K2 ameliorates osteoarthritis by suppressing ferroptosis and extracellular matrix degradation through activation GPX4's dual functions. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 2024 (PubMed: 38759289) [IF=7.5]

Application: WB    Species: Rat    Sample:

Fig. 8. GPX4 inhibitor RSL3 substantially attenuates VK2 protection of chondrocytes. (A-I) WB results for GPX4, NFκB, pMAPK/MAPK, Aggrecan, CollagenⅡ, SOX9, MMP3, MMP13 and Tubulin; (J) Quantitive analysis of relative Glutathione Peroxidase Activity; (K-L) GSH contents and ratio of GSH/GSSG; (M) Quantitative results of MDA content assay.

9). Biochanin A protects against iron overload associated knee osteoarthritis via regulating iron levels and NRF2/System xc-/GPX4 axis. Biomedicine & Pharmacotherapy, 2023 (PubMed: 36379122) [IF=7.5]

10). Cigarette smoke induces the ROS accumulation and iNOS activation through deactivation of Nrf-2/SIRT3 axis to mediate the human bronchial epithelium ferroptosis. Free Radical Biology and Medicine, 2023 (PubMed: 36871899) [IF=7.4]

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