使用敲除细胞株进行验证

Anti-Glutathione Peroxidase 1抗体(ab22604)

概述

  • 产品名称
    Anti-Glutathione Peroxidase 1抗体
    参阅全部 Glutathione Peroxidase 1 一抗
  • 描述
    兔多克隆抗体to Glutathione Peroxidase 1
  • 经测试应用
    适用于: IHC-P, WB, ICC/IF, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Cow, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Glutathione Peroxidase 1.

    (Peptide available as ab25301.)

  • 阳性对照
    • This antibody gave a positive signal in Human, Rat and mouse liver tissue lysates.

性能

应用

Our Abpromise guarantee covers the use of ab22604 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-P 1/300 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration.

靶标

图片

  • ab22604 staining mouse Cor-1 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with TBS/BSA/azide/0.1%Tween 20 and blocked with 1% BSA for 10 minutes at RT. Samples were incubated with primary antibody (1/300) for 2 hours. An diluted (1/1000) Alexa Fluor® 594-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview



  • Predicted band size : 22 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: GPX1 knockout HAP1 cell lysate (20 µg)
    Lane 3: THP1 cell lysate (20 µg)
    Lane 4: HL60 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab22604 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab22604 was shown to recognize Glutathione Peroxidase 1 when Glutathione Peroxidase 1 knockout samples were used, along with additional cross-reactive bands. Wild-type and Glutathione Peroxidase 1 knockout samples were subjected to SDS-PAGE. ab22604 at a concentration of 1µg/ml and ab8245 (loading control to GAPDH) diluted at 1/2000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Image courtesy of Human Protein Atlas

    ab22604 staining Glutathione Peroxidase 1 in Human kidney. The paraffin embedded human kidney tissue was incubated with ab22604 (1/375 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab22604 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • ICC/IF image of ab22604 stained human HepG2 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab22604, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • All lanes : Anti-Glutathione Peroxidase 1 antibody (ab22604) at 1 µg/ml (Blocked with 3% Milk)

    Lane 1 : Liver (Human) Tissue Lysate
    Lane 2 : Liver (Mouse) Tissue Lysate
    Lane 3 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 22 kDa
    Observed band size : 22 kDa
    Additional bands at : 190 kDa,55 kDa,65 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab22604 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-Glutathione Peroxidase 1 antibody (ab22604) at 1 µg/ml

    Lane 1 : Human liver normal tissue lysate
    Lane 2 : Liver (Mouse) Tissue Lysate (ab7935)
    Lane 3 : Human liver normal tissue lysate with Human Glutathione Peroxidase 1 peptide (ab25301) at 1 µg/ml
    Lane 4 : Liver (Mouse) Tissue Lysate (ab7935) with Human Glutathione Peroxidase 1 peptide (ab25301) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 22 kDa
    Observed band size : 22,24 kDa (why is the actual band size different from the predicted?)

    ab22604 detects a band of approximately 22 kDa in human liver lysate. The band detected in the mouse liver lysate runs slightly higher (~ 24kDa). We believe that these bands correspond to Glutathione Peroxidase 1 as they are both blocked by the addition of the immunizing peptide (ab25301).

文献

This product has been referenced in:
  • Magierowski M  et al. Nitric oxide, afferent sensory nerves, and antioxidative enzymes in the mechanism of protection mediated by tricarbonyldichlororuthenium(II) dimer and sodium hydrosulfide against aspirin-induced gastric damage. J Gastroenterol N/A:N/A (2017). Read more (PubMed: 28238019) »
  • Tang J  et al. Potential therapeutic role of punicalagin against mechanical-trauma-induced stress urinary incontinence via upregulation of Nrf2 and TGF-ß1 signaling : Effect of punicalagin on mechanical trauma induced SUI. Int Urogynecol J 28:947-955 (2017). Read more (PubMed: 28168411) »

See all 50 Publications for this product

客户评价及客户问答

Filter by Application

Filter by Species

Filter by Ratings

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cow Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric acid 4%
Permeabilization
No
Specification
kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Miss. Jin Young Park

Verified customer

提交于 Aug 28 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric acid 4%
Permeabilization
No
Specification
kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 04 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Bos taurus Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
20 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Julie Kim

Verified customer

提交于 May 12 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate acid solution
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Miss. Jin Young Park

Verified customer

提交于 Feb 08 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (ovarian cancer (OVCAr-3))
Permeabilization
Yes - Ethanol 70%, 30 min at -20
Specification
ovarian cancer (OVCAr-3)
Username

Abcam user community

Verified customer

提交于 Nov 25 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (kidney)
Antigen retrieval step
None
Permeabilization
No
Specification
kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 15 2016

Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 23 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 18 2016

Application
Western blot
Loading amount
36 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Rat Tissue lysate - whole (Tibia)
Specification
Tibia
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Mr. Helder Fonseca

Verified customer

提交于 Sep 25 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle-AT)
Loading amount
50 µg
Specification
Skeletal muscle-AT
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Dr. G.K Sakellariou

Verified customer

提交于 Sep 28 2012

1-10 of 23 Abreviews

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

注册