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[KD Validated] GPX4 Rabbit mAb (A25009)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of various lysates using [KD Validated] GPX4 Rabbit mAb (A25009) at 1:4000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of lysates from wild type (WT) and GPX4 knockout (KO) U-87 MG cells using [KD Validated] GPX4 Rabbit mAb (A25009) at 1:4000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 20s.

Western blot analysis of lysates from Mouse brain using [KD Validated] GPX4 Rabbit mAb (A25009) at 1:4000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 20s.

Western blot analysis of lysates from wild type (WT) and GPX4 knockdown (KD) U-87 MG cells using [KD Validated] GPX4 Rabbit mAb (A25009) at 1:4000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 15s.

ABclonal:Immunohistochemistry - [KD Validated] GPX4 Rabbit mAb (A25009)

Immunohistochemistry analysis of paraffin-embedded Rat testis tissue using [KD Validated] GPX4 Rabbit mAb (A25009) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse testis tissue using [KD Validated] GPX4 Rabbit mAb (A25009) at a dilution of 1:2000 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - [KD Validated] GPX4 Rabbit mAb (A25009)

Confocal imaging of paraffin-embedded mouse testis using [KD Validated] GPX4 Rabbit mAb (A25009, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 40x.Perform high pressure antigen retrieval with 0.01 M citrate buffer (pH 6.0) prior to IF staining.

Overview

Product name	[KD Validated] GPX4 Rabbit mAb
Catalog No.	A25009
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC65274

Background

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.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 74-197 of human GPX4(NP_002076.2).
Sequence	GKTEVNYTQLVDLHARYAECGLRILAFPCNQFGKQEPGSNEEIKEFAAGYNVKFDMFSKICVNGDDAHPLWKWMKIQPKGKGILGNAIKWNFTKFLIDKNGCVVKRYGPMEEPLVIEKDLPHYF
Gene ID
Swiss Prot
Synonyms	MCSP; SMDS; GPx-4; PHGPx; snGPx; GSHPx-4; snPHGPx; [KD Validated] GPX4
Calculated MW	22kDa
Observed MW	20kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:4000 - 1:24000 IHC-P 1:1000 - 1:4000 IF/ICC 1:200 - 1:1600 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	Jurkat, U-937, U-87 MG, Mouse brain
Cellular location	Cytoplasm, Mitochondrion.
Customer validation	IHC(Mus musculus)