Recombinant
RabMAb

Recombinant Anti-REDD-1/DDIT4 antibody [EPR18716] (ab191871)

Overview

  • Product name

    Anti-REDD-1/DDIT4 antibody [EPR18716]
    See all REDD-1/DDIT4 primary antibodies
  • Description

    Rabbit monoclonal [EPR18716] to REDD-1/DDIT4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human REDD-1/DDIT4 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9NX09

  • Positive control

    • WB: HeLa and MCF7 treated with 0.5 mM CoCl2 for 6 hours whole cell lysates. ICC/IF: HeLa cells treated with 0.5mM CoCl2 for 6 hours. IP: HeLa treated with 0.5mM CoCl2 for 6 hours whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab191871 in the following tested applications.

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

Application Abreviews Notes
IP 1/50.
ICC/IF 1/250.
WB 1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 25 kDa).

Target

  • Function

    Inhibits cell growth by regulating the TOR signaling pathway upstream of the TSC1-TSC2 complex and downstream of AKT1. Promotes neuronal cell death.
  • Tissue specificity

    Broadly expressed, with lowest levels in brain, skeletal muscle and intestine. Up-regulated in substantia nigra neurons from Parkinson disease patients (at protein level).
  • Sequence similarities

    Belongs to the DDIT4 family.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • DDIT4 antibody
    • DDIT4_HUMAN antibody
    • Dig2 antibody
    • DNA damage inducible transcript 4 antibody
    • DNA damage inducible transcript 4 protein antibody
    • DNA damage-inducible transcript 4 protein antibody
    • FLJ20500 antibody
    • HIF 1 responsive protein RTP801 antibody
    • HIF 1 responsive RTP801 antibody
    • HIF-1 responsive protein RTP801 antibody
    • Protein regulated in development and DNA damage response 1 antibody
    • REDD-1 antibody
    • REDD1 antibody
    • RTP801 antibody
    see all

Images

  • All lanes : Anti-REDD-1/DDIT4 antibody [EPR18716] (ab191871) at 1/1000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 0.5 mM CoCl2 for 6 hours whole cell lysate
    Lane 3 : Untreated MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) treated with 0.5 mM CoCl2 for 6 hours whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 25 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 100 methanol-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, untreated or treated with CoCl2 (0.5 mM) for 6 hours, labeling REDD-1/DDIT4 with ab191871 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line. The expression increased after treatment with CoCl2 (0.5 mM) for 6 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).


    The negative controls are as follows:
    -ve control 1: ab191871 at 1/250 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • REDD-1/DDIT4 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 0.5mM CoCl2 for 6 hours whole cell lysate with ab191871 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab191871 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa treated with 0.5mM CoCl2 for 6h whole cell lysate, 10µg (Input).

    Lane 2: ab191871 IP in HeLa treated with 0.5mM CoCl2 for 6h whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal- Isotype Control (ab172730) instead of ab191871 in HeLa treated with 0.5mM CoCl2 for 6h whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 1 second.

References

ab191871 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Cell lysate - whole cell (skeletal muscle lysates)
Gel Running Conditions
Reduced Denaturing (4-15% TGX Stain-free)
Loading amount
20 µg
Specification
skeletal muscle lysates
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

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Verified customer

Submitted Apr 19 2018

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