Overview

  • Product name

    Anti-Calbindin antibody [EP3478]
    See all Calbindin primary antibodies
  • Description

    Rabbit monoclonal [EP3478] to Calbindin
  • Host species

    Rabbit
  • Specificity

    We do not guarantee IHC-P for mouse and rat.

    We do not guarantee IHC-Fr for human.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IHC-Fr, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Calbindin aa 50-150 (internal sequence). The exact sequence is proprietary.

  • Positive control

    • WB: Recombinant Human Calbindin protein (ab117207), fetal brain, fetal cerebellum, fetal hypothalamus and fetal pituitary lysates, SH-SY5Y, Human kidney, Mouse brain, Mouse kidney, and Rat brain lysates. IHC-P: Human cerebrum and Human brain tissue. IHC-Fr: Mouse and Rat cerebellum tissues. ICC/IF: SH-SY5Y cells. FC: SH-SY5Y cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab108404 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 1/1000 - 1/10000. Predicted molecular weight: 30 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

We do not guarantee IHC-P for mouse and rat.

See IHC antigen retrieval protocols

For unpurified use at 1/100 - 1/250. 

ICC/IF 1/150.

For unpurified use at 1/100

IHC-Fr 1/500.

We do not guarantee IHC-Fr for human

Flow Cyt 1/150.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Buffers cytosolic calcium. May stimulate a membrane Ca(2+)-ATPase and a 3',5'-cyclic nucleotide phosphodiesterase.
    • Sequence similarities

      Belongs to the calbindin family.
      Contains 5 EF-hand domains.
    • Domain

      This protein has four functional calcium-binding sites; potential sites II and VI have lost affinity for calcium.
    • Information by UniProt
    • Database links

    • Alternative names

      • avian-type antibody
      • CAB27 antibody
      • CALB 1 antibody
      • CALB antibody
      • CALB1 antibody
      • CALB1_HUMAN antibody
      • Calbindin 1 28kDa antibody
      • Calbindin antibody
      • Calbindin D28 antibody
      • D 28K antibody
      • D-28K antibody
      • D28K antibody
      • OTTHUMP00000166027 antibody
      • OTTHUMP00000225441 antibody
      • RTVL H protein antibody
      • Vitamin D dependent calcium binding protein antibody
      • Vitamin D dependent calcium binding protein avian type antibody
      • Vitamin D-dependent calcium-binding protein antibody
      see all

    Images

    • All lanes : Anti-Calbindin antibody [EP3478] (ab108404) at 1/2000 dilution (Purified)

      Lane 1 : Human fetal brain lysates
      Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
      Lane 3 : Human kidney lysates
      Lane 4 : Mouse brain lysates
      Lane 5 : Mouse kidney lysates
      Lane 6 : Rat brain lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 30 kDa
      Observed band size: 30 kDa

    • Immunohistochemistry (Frozen) analysis of mouse cerebellum tissue sections labeling Calbindin with purified ab108404 at 1/500 dilution (3.0 µg/ml). The tissue section was fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. DAPI (blue) was used as nuclear counterstain.
      Secondary antibody only control: PBS instead of the primary antibody
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling Calbindin with purified ab108404 at 1/1000 dilution (1.49 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • All lanes : Anti-Calbindin antibody [EP3478] (ab108404) at 1/1000 dilution

      Lane 1 : Fetal brain lysate
      Lane 2 : Fetal cerebellum lysate
      Lane 3 : Fetal hypothalamus lysate
      Lane 4 : Fetal pituitary lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 30 kDa

    • Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Calbindin with purified ab108404 at 1/150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
    • Immunohistochemistry (Frozen) analysis of rat cerebellum tissue sections labeling Calbindin with purified ab108404 at 1/500 dilution (3.0 µg/ml). The tissue section was fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. DAPI (blue) was used as nuclear counterstain.
      Secondary antibody only control: PBS instead of the primary antibody
    • Immunocytochemistry/ Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Calbindin with purified ab108404 at 1/150 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Immunohistochemical analysis of mouse frozen cerebellum tissue sections with ab108404 labelling Calbindin at 1/500. Sections were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).  The image shows specific staining on Purkinje cells of mouse cerebellum.

      Secondary antibody only was used as negative control and is shown in the inset.

    • ab108404 showing positive staining in Normal kidney tissue.

    • Immunocytochemistry/Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma) cells labelling Calbindin with ab108404 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at dilution of 1/200. Nuclei were counterstained with DAPI (blue).

      Confocal image showing nuclear and cytoplasmic staining on Neuro2a cultured with 2% FBS.

    • ab108404, at 1/100 dilution, staining Calbindin in paraffin-embedded Human brain tissue by Immunohistochemistry.
    • ab108404 showing negative staining in Normal heart tissue.

    • ab108404 showing negative staining in Normal colon tissue.

    • ab108404 showing negative staining in Normal liver tissue.

    References

    This product has been referenced in:

    • Qin J  et al. Early Forebrain Neurons and Scaffold Fibers in Human Embryos. Cereb Cortex N/A:N/A (2019). Read more (PubMed: 31298263) »
    • Dietrich P  et al. Elimination of huntingtin in the adult mouse leads to progressive behavioral deficits, bilateral thalamic calcification, and altered brain iron homeostasis. PLoS Genet 13:e1006846 (2017). Read more (PubMed: 28715425) »
    See all 4 Publications for this product

    Customer reviews and Q&As

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    1-2 of 2 Abreviews

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rhesus monkey Tissue sections (macaque retina)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate pH 6
    Permeabilization
    No
    Specification
    macaque retina
    Blocking step
    Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Nov 06 2018

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Fukomys anselli Tissue sections (Retina)
    Permeabilization
    Yes - Triton X-100 in Blocking Solution
    Specification
    Retina
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
    Fixative
    Davidson's Fixative

    Abcam user community

    Verified customer

    Submitted Jul 28 2017

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