Overview

  • Product nameAnti-Hephaestin antibody
    See all Hephaestin primary antibodies
  • Description
    Rabbit polyclonal to Hephaestin
  • Tested applicationsSuitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Hephaestin.

    (Peptide available as ab109074.)

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

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

Anti-Hephaestin antibody images

  • 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

  • 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
  • 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 for Anti-Hephaestin antibody (ab108003)

ab108003 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Sample Rat Cell (Hippocampal culture)
Specification Hippocampal culture
Permeabilization Yes - 0.1% TritonX in 0.1% PBS
Fixative Paraformaldehyde
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Jul 16 2013

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