WB
Western blot analysis of GPX4 on different lysates with Rabbit anti-GPX4 antibody at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: HepG2 cell lysate (20 µg/Lane) Lane 4: U-937 cell lysate (20 µg/Lane) Lane 5: HEK-293 cell lysate (20 µg/Lane) Lane 6: Mouse testis tissue lysate (20 µg/Lane) Lane 7: Rat testis tissue lysate (20 µg/Lane) Lane 8: Mouse kidney tissue lysate (20 µg/Lane) Lane 9: Rat kidney tissue lysate (20 µg/Lane) Lane 10: Mouse brain tissue lysate (20 µg/Lane) Lane 11: Rat brain tissue lysate (20 µg/Lane) Predicted band size: 22 kDa Observed band size: 22 kDa Exposure time: 2 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 at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-GPX4 antibody at 1/1,000 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 at 1/1,000 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.ICC/IF
Immunocytochemistry analysis of HEK-293 cells labeling GPX4 with Rabbit anti-GPX4 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-GPX4 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HEK-293 cells labeling GPX4. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | GPX4 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human GPX4 aa 23-72 / 197. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:10000 IHC:1:200-1:1000 ICC:1:50-1:200 FC:1:10000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | GPX4 |
|---|---|
| Gene Synonyms | MCSP SMDS GPx-4 PHGPx snGPx GSHPx-4 snPHGPx |
| Gene Full Name | glutathione peroxidase 4 |
| Gene Summary | The protein encoded by this gene belongs to the glutathione peroxidase family, members of which catalyze the reduction of hydrogen peroxide, organic hydroperoxides and lipid hydroperoxides, and thereby protect cells against oxidative damage. Several isozymes of this gene family exist in vertebrates, which vary in cellular location and substrate specificity. This isozyme has a high preference for lipid hydroperoxides and protects cells against membrane lipid peroxidation and cell death. It is also required for normal sperm development; thus, it has been identified as a 'moonlighting' protein because of its ability to serve dual functions as a peroxidase, as well as a structural protein in mature spermatozoa. Mutations in this gene are associated with Sedaghatian type of spondylometaphyseal dysplasia (SMDS). This isozyme is also a selenoprotein, containing the rare amino acid selenocysteine (Sec) at its active site. Sec is encoded by the UGA codon, which normally signals translation termination. The 3' UTRs of selenoprotein mRNAs contain a conserved stem-loop structure, designated the Sec insertion sequence (SECIS) element, that is necessary for the recognition of UGA as a Sec codon, rather than as a stop signal. Transcript variants resulting from alternative splicing or use of alternate promoters have been described to encode isoforms with different subcellular localization. [provided by RefSeq, Dec 2018] |
| Alternative Names | Glutathione peroxidase 4 antibody GPX 4 antibody GPX-4 antibody GPX4 antibody GPX4_HUMAN antibody GSHPx-4 antibody MCSP antibody mitochondrial antibody PHGPx antibody Phospholipid hydroperoxidase antibody Glutathione peroxidase 4 antibody GPX 4 antibody GPX-4 antibody GPX4 antibody GPX4_HUMAN antibody GSHPx-4 antibody MCSP antibody mitochondrial antibody PHGPx antibody Phospholipid hydroperoxidase antibody Phospholipid hydroperoxide glutathione peroxidase antibody Phospholipid hydroperoxide glutathione peroxidase mitochondrial antibody snGPx antibody snPHGPx antibody Sperm nucleus glutathione peroxidase antibody |
| Molecular Weight(MW) | Predicted band size: 22 kDa |
| Cellular Localization | Mitochondrion, Cytoplasm. |
WB
Western blot analysis of GPX4 on different lysates with Rabbit anti-GPX4 antibody at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: HepG2 cell lysate (20 µg/Lane) Lane 4: U-937 cell lysate (20 µg/Lane) Lane 5: HEK-293 cell lysate (20 µg/Lane) Lane 6: Mouse testis tissue lysate (20 µg/Lane) Lane 7: Rat testis tissue lysate (20 µg/Lane) Lane 8: Mouse kidney tissue lysate (20 µg/Lane) Lane 9: Rat kidney tissue lysate (20 µg/Lane) Lane 10: Mouse brain tissue lysate (20 µg/Lane) Lane 11: Rat brain tissue lysate (20 µg/Lane) Predicted band size: 22 kDa Observed band size: 22 kDa Exposure time: 2 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 at 1/10,000 dilution and competitor's antibody at 1/10,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-GPX4 antibody at 1/1,000 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 at 1/1,000 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.
ICC/IF
Immunocytochemistry analysis of HEK-293 cells labeling GPX4 with Rabbit anti-GPX4 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-GPX4 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 ( red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HEK-293 cells labeling GPX4. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:10000 IHC:1:200-1:1000 ICC:1:50-1:200 FC:1:10000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
