Overview

  • Product name
  • Description
    Rabbit polyclonal to Calbindin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, WB, IHC-P, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Rabbit, Horse, Chicken, Hamster, Cow
  • Immunogen

    Synthetic peptide within Human Calbindin aa 150-250. The exact sequence is proprietary.
    Database link: P05937

  • Positive control
    • Rat brain

Properties

Applications

Our Abpromise guarantee covers the use of ab25085 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 Use a concentration of 5 µg/ml.
WB Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 28 kDa.
IHC-P 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 - 5 µg/ml.

Target

Images

  • Calbindin was immunoprecipitated using 0.5mg Rat Brain tissue extract, 5µg of Rabbit polyclonal to Calbindin 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, Rat Brain tissue extract 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab25085.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).<

    Band: 28kDa; Calbindin

  • ab25085 staining human Calbindin D28 in Human cerebellum by immunohistochemistry formalin fixed, paraffin embedded.
  • ICC/IF image of ab25085 stained PC12 cells. The cells were 100% methanol fixed (5 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 (ab25085, 1ug/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-rabbit 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 1.43uM.

  • Anti-Calbindin antibody (ab25085) at 1 µg/ml + Brain (Rat) Tissue Lysate 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: 28 kDa
    Observed band size: 30 kDa (why is the actual band size different from the predicted?)
    Additional bands at: 67 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes

References

This product has been referenced in:
  • Bannerman P  et al. Brain Nat8l Knockdown Suppresses Spongiform Leukodystrophy in an Aspartoacylase-Deficient Canavan Disease Mouse Model. Mol Ther 26:793-800 (2018). Read more (PubMed: 29456021) »
  • Noreldin AE  et al. Immunohistochemical localization of osteoblast activating peptide in the mouse kidney. Acta Histochem N/A:N/A (2018). Read more (PubMed: 29534834) »

See all 14 Publications for this product

Customer reviews and Q&As

I apologize once again for my mistake in shipping you the wrong antibody. The new antibody ab49899 will be shipped on Monday and will arrive on Tuesday. Your order number is ******

I am sorry that you were having issues with ab25085 in immuno staining. I have sent you the replacement antibody ab49889, which is only listed as recognizing calbindin and also is raised against a different sequence compare to ab2085. Your new order n...

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

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