GPX4 Antibody (YA399)

GPX4 Antibody (YA399) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to GPX4.

GPX4 is an essential antioxidant peroxidase that directly reduces phospholipid hydroperoxides, including those incorporated in membranes and lipoproteins (By similarity). It also reduces cholesterol hydroperoxide and thymine hydroperoxide (By similarity), playing a key role in protecting cells from oxidative damage by preventing membrane lipid peroxidation (By similarity). GPX4 is required to inhibit ferroptosis, a non-apoptotic cell death caused by iron-dependent accumulation of lipid reactive oxygen species (PubMed:24439385). The presence of selenocysteine (Sec) versus cysteine (Cys) at the active site is critical for life, as it provides resistance to overoxidation and prevents ferroptosis (By similarity). Sec is also essential for the survival of parvalbumin-positive interneurons, thereby preventing fatal epileptic seizures (By similarity). GPX4 may protect cells from the toxicity of ingested lipid hydroperoxides (By similarity) and is indispensable for normal sperm development, male fertility (By similarity), and photoreceptor cell maturation and survival (By similarity). In T-cell responses to viral and parasitic infections, GPX4 prevents ferroptosis and supports T-cell expansion (By similarity). Additionally, it functions as a glutathione peroxidase in platelets during arachidonic acid metabolism (PubMed:11115402) and reduces hydroperoxy ester lipids formed by 15-lipoxygenase, downregulating the cellular 15-lipoxygenase pathway (By similarity). GPX4 can also reduce fatty acid-derived hydroperoxides (PubMed:11115402, PubMed:36608588) and small soluble hydroperoxides such as H2O2, cumene hydroperoxide, and tert-butyl hydroperoxide (PubMed:17630701, PubMed:36608588).

Free

P36969

Predicted band size: 22 kDa; Observed band size: 20 kDa

Protein A affinity purified.

Mitochondrion, Cytoplasm.

Non-conjugated

Unmodified

AB_3102526

Signal Transduction

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat

WB: 1:500-1:2000; IHC-P: 1:50-1:500; ICC: 1:50-1:200; FC: 1:1000

• 
  Western blot analysis of extracts from HEK293 (lane 2(40μg) and HEK293 (lane 3(20μg)using GPX4 (HY-P80450) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% defatted milk powder in TBST for 2 hour at room temperature. The primary antibody (HY-P80450, 1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% defatted milk powder in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of 293 cells labeling GPX4 with GPX4 Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of C6 cells labeling GPX4 with GPX4 Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) 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. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded mouse testis tissue using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1:1000) in 4℃ overnight. 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 testis tissue using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1:1000) in 4℃ overnight. 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 1X10^6 HEK293 cells labeling GPX4 Antibody (HY-P80450, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1μg/mL dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1/1000) in 4℃ overnight. 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 using GPX4 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80450, 1/1000) in 4℃ overnight. 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.
• 
  Immunocytochemistry analysis of Hela cells labeling GPX4 with GPX4Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) 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. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of HepG2 cells labeling GPX4 with GPX4 Antibody (HY-P80450) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GPX4 Antibody (HY-P80450)at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002,Green) 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. The Nuclear counterstain was DAPI (Blue).

WB, IHC-P, ICC/IF, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human GPX4.AA range:23-72.