GPX4 Recombinant Rabbit Monoclonal Antibody [JU11-31]

Specification

50μl

100μl

Quota

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BIO-STATION LIMITED

Contact: Sanry CHEN

Email: info@bio-station.net

Phone: 852-35650223

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Catalog# ET1706-45

GPX4 Recombinant Rabbit Monoclonal Antibody [JU11-31]

Application

WB
IHC-P
IF-Cell
FC

Reactivity

Human
Mouse
Rat
Zebrafish

Predicted reactivity

Pig
Cynomolgus monkey

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

GPX4 Recombinant Rabbit Monoclonal Antibody [JU11-31]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human GPX4 aa 23-72 / 197.

Species Reactivity

Human, Mouse, Rat, Zebrafish (Predicted: Pig, Cynomolgus monkey)

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 22 kDa

Positive Control

HeLa cell lysate, A549 cell lysate, HepG2 cell lysate, U-937 cell lysate, HEK-293 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, Mouse kidney tissue lysate, Rat kidney tissue lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Hela-si NT cell lysate, Hela-si GPX4 cell lysate, zebrafish tissue lysates, HEK-293, human kidney tissue, mouse kidney tissue, mouse testis tissue, C6.

Conjugation

unconjugated

Clone Number

JU11-31

RRID

AB_3070665

Product Features

Form

Liquid

Concentration

1ug/ul

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

WB
1:10,000

IHC-P
1:1,000-1:5,000

IF-Cell
1:50-1:200

FC
1:1,000

Applications in Publications

WB	See 82 publications below
IHC	See 11 publications below
IF	See 5 publications below
ICC	See 1 publications below

Species in Publications

Mouse	See 45 publications below
Human	See 26 publications below
Rat	See 12 publications below
Zebrafish	See 3 publications below
Yellow catfish	See 1 publications below
Porcine	See 1 publications below
mice	See 1 publications below
Bovine	See 1 publications below
Fish	See 1 publications below
rat	See 1 publications below
Rutin	See 1 publications below

Target

Function

Glutathione peroxidase (GPx) enzymes are generally selenium-containing tetrameric glycoproteins that help prevent lipid peroxidation of cell membranes. GPx enzymes reduce lipid hydroperoxides to alcohols, and reduce free hydrogen peroxide to water. GPx members are among the few proteins known in higher vertebrates to contain selenocysteine, which occurs at the active site of glutathione peroxidase and is coded by the nonsense (stop) codon TGA. There are eight GPx homologs (GPx-1-8). GPx-1, Gpx-2 and Gpx-3 exist as homotetramers. Gpx-4 has a high tendancy to form high molecular weight oligomers.

Background References

1. Luna-Sánchez M et al. CoQ deficiency causes disruption of mitochondrial sulfide oxidation, a new pathomechanism associated with this syndrome. EMBO Mol Med 9:78-95 (2017).

2. Kagan VE et al. Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis. Nat Chem Biol 13:81-90 (2017).

Sequence Similarity

Belongs to the glutathione peroxidase family.

Tissue Specificity

Present primarily in testis. Expressed in platelets (at protein level).

Subcellular Location

Mitochondrion, Cytoplasm.

UNIPROT

SwissProt: P36969 Human

SwissProt: O70325 Mouse

SwissProt: P36970 Rat

Synonyms

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

Expand

Images

Western blot analysis of GPX4 on different lysates with Rabbit anti-GPX4 antibody (ET1706-45) 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: 20 kDa
Exposure time: 2 minutes; 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 (ET1706-45) 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 (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of GPX4 on zebrafish tissue lysates with Rabbit anti-GPX4 antibody (ET1706-45) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 22 kDa
Observed band size: 17 kDa

Exposure time: 2 minutes;

12% 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 (ET1706-45) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)

Western blot analysis of GPX4 on different lysates with Rabbit anti-GPX4 antibody (ET1706-45) at 1/10,000 dilution.

Lane 1: Hela-si NT cell lysate, 10 µg/Lane
Lane 2: Hela-si GPX4 cell lysate, 10 µg/Lane

Predicted band size: 22 kDa
Observed band size: 20,22 kDa
Exposure time: 1 minutes 40 seconds;
4-20% SDS-PAGE gel.

ET1706-45 was shown to specifically react with GPX4 in Hela-si NT cells. Weakened band was observed when Hela-si GPX4 sample was tested. Hela-si NT and Hela-si GPX4 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1706-45, 1/10,000) and Loading control antibody (Rabbit anti-GAPDH , ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HEK-293 cells labeling GPX4 with Rabbit anti-GPX4 antibody (ET1706-45) at 1/200 dilution and competitor's antibody at 1/200 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 (ET1706-45) at 1/200 dilution and competitor's antibody at 1/200 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 (M1305-2, 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 kidney tissue with Rabbit anti-GPX4 antibody (ET1706-45) 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 (ET1706-45) 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.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-GPX4 antibody (ET1706-45) 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 (ET1706-45) 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.
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-GPX4 antibody (ET1706-45) at 1/5,000 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 (ET1706-45) at 1/5,000 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.
Immunocytochemistry analysis of C6 cells labeling GPX4 with Rabbit anti-GPX4 antibody (ET1706-45) 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 (ET1706-45) 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 (M1305-2, 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.
Flow cytometric analysis of HEK-293 cells labeling GPX4.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1706-45, 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 (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).