Overview

  • Product nameAnti-Calbindin antibody
    See all Calbindin primary antibodies
  • Description
    Rabbit polyclonal to Calbindin
  • Tested applicationsSuitable 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:

    MCGKEFNKAFELYDQ

    , corresponding to amino acids 185-199 of Human Calbindin

  • Positive control
    • Rat brain

Properties

Associated products

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

Anti-Calbindin antibody 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

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 28 kDa


    Exposure time : 30 seconds

References for Anti-Calbindin antibody (ab25085)

This product has been referenced in:
  • Kim JH  et al. Distinctive genetic activity pattern of the human dental pulp between deciduous and permanent teeth. PLoS One 9:e102893 (2014). IHC-P ; Human . Read more (PubMed: 25047033) »
  • Anders C  et al. Ex vivo modeling of chemical synergy in prenatal kidney cystogenesis. PLoS One 8:e57797 (2013). IHC-P ; Mouse . Read more (PubMed: 23554868) »

See all 7 Publications for this product

Product Wall

Application Western blot
Sample Chicken Tissue lysate - whole (Duodenum)
Gel Running Conditions Reduced Denaturing (gradient 4-20%)
Loading amount 10 µg
Specification Duodenum
Blocking step Odyssey® Blocking Buffer (PBS) as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Mr. Aurelien Brionne

Verified customer

Submitted Jul 28 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step None
Sample Human Tissue sections (tooth)
Specification tooth
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 17 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Kidney)
Specification Kidney
Fixative Acetone
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Mar 30 2012

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...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Brain)
Loading amount 20 µg
Specification Brain
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 13 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization Yes - Triton X-100
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Kidney)
Specification Kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 1%
Username

Abcam user community

Verified customer

Submitted Jul 31 2007

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