Overview

  • Product name

  • Description

    Rabbit polyclonal to HIGD2A
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse
    Predicted to work with: Rat, Horse, Chimpanzee, Macaque monkey, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Mouse HIGD2A aa 1-100 conjugated to keyhole limpet haemocyanin.
    Database link: Q9CQJ1

  • Positive control

    • This antibody gave a positive signal in Mouse Heart and Kidney tissue lysates. IF/ICC: HepG2 cell line.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab135399 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 13 kDa (predicted molecular weight: 11 kDa).

Target

  • Cellular localization

    Membrane; Multi-pass membrane protein
  • Database links

  • Alternative names

    • HIG1 domain family member 2A antibody
    • HIG1 hypoxia inducible domain family, member 2A antibody
    • Higd2a antibody

Images

  • ICC/IF image of ab135399 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab135399, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) HeLa and MCF7 cells at 5µg/ml.

  • All lanes : Anti-HIGD2A antibody (ab135399) at 1 µg/ml

    Lane 1 : Heart (Mouse) Tissue Lysate
    Lane 2 : Kidney (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 11 kDa
    Observed band size: 13 kDa
    why is the actual band size different from the predicted?


    Exposure time: 2 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 5% Bovine Serum Albumin before being incubated with ab135399 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

ab135399 has not yet been referenced specifically in any publications.

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