• Product nameAnti-Hippocalcin antibody
    See all Hippocalcin primary antibodies
  • Description
    Rabbit polyclonal to Hippocalcin
  • Tested applicationsSuitable for: WB, IHC-P, IHC (PFA fixed), ICC/IF, IHC-Fr, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Chicken, Cow
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Rat Hippocalcin.

    (Peptide available as ab25848.)

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Mouse Brain; Mouse Brain Day 0; Human Cerebellum. This antibody gave a positive signal in the following Formaldehyde fixed cell line: SKNSH.



Our Abpromise guarantee covers the use of ab24560 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
IHC-P 1/350. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC (PFA fixed) Use a concentration of 0.2 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.


Anti-Hippocalcin antibody images

  • All lanes : Anti-Hippocalcin antibody (ab24560) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate
    Lane 4 : Mouse brain tissue lysate - total protein (0 days) (ab7188)

    Lysates/proteins at 20 µg per lane.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 23 kDa
    Observed band size : 23 kDa
    Additional bands at : 14 kDa (possible cleavage fragment),42 kDa,52 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes
  • Immuofluorescent staining for Hippocalcin in the rat hippocampus using Rabbit polyclonal to Hippocalcin (ab24560). This figure is a montage of pictures acquired with a X10 objective and shows expected abundance of staining in parts of the hippocampus such as the CA1 and CA3 and an absence of staining (as expected) in the cortex and the corpus callosum. ab24560 was used at 1/1000 (0.2µg/ml) incubated overnight at room temperature. Secondary antibody used was anti-rabbit Alexa Fluor 488 at 1/1000 incubated for 2 hours at room temperature. Rat brain tissue was perfusion fixed with 4% PFA followed by overnight post-fixation in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut on a cyrostat and immunohistochemistry performed by the 'free floating' technique.

  • Image courtesy of Human Protein Atlas
    ab24560 staining Hippocalcin in human cerebellum. Paraffin embedded tissue was cut into 4µm sections and incubated with ab24560 (1/350 dilution) for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab24560 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
  • Hippocalcin was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to Hippocalcin 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, Mouse Brain whole tissue lysate 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 ab24560.
    Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
    Band: 23kDa: Hippocalcin; non specific - 60kDa: We are unsure as to the identity of this extra band.
  • ICC/IF image of ab24560 stained SKNSH 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 ab24560 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.

  • All lanes : Anti-Hippocalcin antibody (ab24560) at 1 µg/ml

    Lane 1 : Marker
    Lane 2 : Zebrafish brain homogenate (20ug)
    Lane 3 : Mouse brain homogenate (20ug)

    Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 23 kDa
    Observed band size : 23 kDa

    Exposure time : 1 minute

References for Anti-Hippocalcin antibody (ab24560)

This product has been referenced in:
  • Magnani D  et al. Gli3 Controls Subplate Formation and Growth of Cortical Axons. Cereb Cortex : (2012). Read more (PubMed: 22903314) »
  • Kathiresan T  et al. A protein interaction network for the large conductance Ca(2+)-activated K(+) channel in the mouse cochlea. Mol Cell Proteomics 8:1972-87 (2009). Read more (PubMed: 19423573) »

See all 3 Publications for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Sections of rat brain)
Specification Sections of rat brain
Fixative Paraformaldehyde

Dr. Sophie Pezet

Verified customer

Submitted May 17 2006