Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Hephaestin antibody (ab108003)

Key features and details

  • Rabbit polyclonal to Hephaestin
  • Suitable for: ICC/IF, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Hephaestin antibody

  • Description

    Rabbit polyclonal to Hephaestin

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Dog

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • This antibody gave a positive signal in WI38 Whole Cell Lysate.

  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab108003 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 1/250. Detects a band of approximately 130 kDa (predicted molecular weight: 130 kDa).

Target

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)
    Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)This image is courtesy of an abreview submitted by Dr. Ruma Raha-Chowdhury, Cambridge University, United Kingdom.

    ICC/IF image of Hephaestin staining on rat hippocampal culture  using ab108003 (1:200). The cells were fixed with paraformaldehyde and permeabilized using 0.1% TritonX in 0.1% PBS. The cells were blocked using 10 % donkey serum for 1 hour at 24°C. ab108003 was diluted 1:200 using 0.1% TritonX with 0.1x PBS- 10% Donkeys and incubated with the slides at 4°C for 24 hours. The secondary antibody used was donkey polyclonal against rabbit IgG conjugated to Alexa Fluor 488 (1:1000)

    See Abreview

  • Western blot - Anti-Hephaestin antibody (ab108003)
    Western blot - Anti-Hephaestin antibody (ab108003)
    Anti-Hephaestin antibody (ab108003) at 1/250 dilution + WI-38 whole cell lysate (ab3960) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 130 kDa
    Observed band size: 130 kDa


    Exposure time: 20 minutes
  • Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)
    Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)
    ICC/IF image of ab108003 stained MCF7 cells. The cells were 4% PFA 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 (ab108003, 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% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.

References (1)

Publishing research using ab108003? Please let us know so that we can cite the reference in this datasheet.

ab108003 has been referenced in 1 publication.

  • Kenawi M  et al. Ceruloplasmin deficiency does not induce macrophagic iron overload: lessons from a new rat model of hereditary aceruloplasminemia. FASEB J 33:13492-13502 (2019). PubMed: 31560858

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)
    Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003) This image is courtesy of an abreview submitted by Dr. Ruma Raha-Chowdhury, Cambridge University, United Kingdom.

    ICC/IF image of Hephaestin staining on rat hippocampal culture  using ab108003 (1:200). The cells were fixed with paraformaldehyde and permeabilized using 0.1% TritonX in 0.1% PBS. The cells were blocked using 10 % donkey serum for 1 hour at 24°C. ab108003 was diluted 1:200 using 0.1% TritonX with 0.1x PBS- 10% Donkeys and incubated with the slides at 4°C for 24 hours. The secondary antibody used was donkey polyclonal against rabbit IgG conjugated to Alexa Fluor 488 (1:1000)

    See Abreview

  • Western blot - Anti-Hephaestin antibody (ab108003)
    Western blot - Anti-Hephaestin antibody (ab108003)
    Anti-Hephaestin antibody (ab108003) at 1/250 dilution + WI-38 whole cell lysate (ab3960) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 130 kDa
    Observed band size: 130 kDa


    Exposure time: 20 minutes
  • Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)
    Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)
    ICC/IF image of ab108003 stained MCF7 cells. The cells were 4% PFA 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 (ab108003, 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% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.

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