概述

  • 产品名称Anti-Glutaredoxin 1抗体
    参阅全部 Glutaredoxin 1 一抗
  • 描述
    兔多克隆抗体to Glutaredoxin 1
  • 经测试应用适用于: IHC-P, IP, ICC/IF, WBmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse, Rat, Rabbit, Cow, Pig
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Glutaredoxin 1.

    (Peptide available as ab45952.)

  • 阳性对照
    • This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line)

性能

应用

Our Abpromise guarantee covers the use of ab45953 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 Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
WB 1/250. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).

靶标

Anti-Glutaredoxin 1 antibody 图像

  • Anti-Glutaredoxin 1 antibody (ab45953) at 1/250 dilution + Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 12 kDa
    Observed band size : 12 kDa
    Additional bands at : 45 kDa,70 kDa. We are unsure as to the identity of these extra bands.
  • ICC/IF image of ab45953 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab45953, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and 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).

  • IHC image of ab45953 staining in cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45953, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Glutaredoxin 1 antibody (ab45953) at 1/250 dilution

    Lane 1 : Recombinant Human Glutaredoxin 1 protein (ab86987) at 0.01 µg
    Lane 2 : Recombinant Human Glutaredoxin 1 protein (ab86987) at 0.001 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 1 minute
  • Glutaredoxin 1 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to Glutaredoxin 1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab45953.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 12kDa; Glutaredoxin 1.

Anti-Glutaredoxin 1 antibody (ab45953)参考文献

This product has been referenced in:
  • Liu X  et al. The novel triterpenoid RTA 408 protects human retinal pigment epithelial cells against H2O2-induced cell injury via NF-E2-related factor 2 (Nrf2) activation. Redox Biol 8:98-109 (2016). Human . Read more (PubMed: 26773873) »
  • Mailloux RJ  et al. Crucial yet divergent roles of mitochondrial redox state in skeletal muscle vs. brown adipose tissue energetics. FASEB J 26:363-75 (2012). WB . Read more (PubMed: 21940996) »

See all 5 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (rat heart)
Gel Running Conditions Reduced Denaturing (15% resolving gel)
Loading amount 25 µg
Specification rat heart
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Abcam user community

Verified customer

提交于 Mar 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Human Lens epithelial cells)
Loading amount 25 µg
Specification Human Lens epithelial cells
Gel Running Conditions Reduced Denaturing (12% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Abcam user community

Verified customer

提交于 Dec 22 2011

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