Overview

  • Product name

    Anti-Calbindin antibody [EPR22698-236]
    See all Calbindin primary antibodies
  • Description

    Rabbit monoclonal [EPR22698-236] to Calbindin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-Fr, IHC-P, WB, IPmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Mouse Calbindin aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P12658

  • Positive control

    • WB: Mouse brain and cerebellum lysates; rat brain lysate. IP: Rat brain lysate; mouse brain lysate. IHC-P: Rat cerebellum tissue; mouse cerebellum tissue. IHC-Fr: Rat cerebellum tissue; mouse cerebellum tissue.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab229915 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
IHC-Fr 1/500.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
IP 1/30.
  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.
  • 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

    • Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse cerebellum tissue labeling Calbindin with ab229915 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on molecular layer and purkinje cells of mouse cerebellum (PMID: 20130198) is observed. The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    • Immunohistochemical analysis of frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized rat cerebellum tissue labeling Calbindin with ab229915 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution (green). Positive staining on molecular layer and purkinje cells of rat cerebellum (PMID: 20130198) is observed. The nuclear counter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    • Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling Calbindin with ab229915 at 1/4000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Purkinje cells of rat cerebellum (PMID: 17507571) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

      The section was incubated with ab229915 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    • Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling Calbindin with ab229915 at 1/4000 dilution, followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Strong positive staining on Purkinje cells of mouse cerebellum (PMID: 17507571) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

      The section was incubated with ab229915 for 30 mins at room temperature.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    • Calbindin was immunoprecipitated from 0.35 mg of mouse brain lysate with ab229915 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229915 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.

      Lane 1: Mouse brain lysate 10 μg (Input).
      Lane 2: ab229915 IP in mouse brain lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229915 in mouse brain lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 5 seconds.

    • Calbindin was immunoprecipitated from 0.35 mg of rat brain lysate with ab229915 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229915 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.

      Lane 1: Rat brain lysate 10 μg (Input). 
      Lane 2: ab229915 IP in rat brain lysate. 
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab229915 in rat brain lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 30 seconds.

    • All lanes : Anti-Calbindin antibody [EPR22698-236] (ab229915) at 1/1000 dilution

      Lane 1 : Mouse brain lysate
      Lane 2 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
      Lane 3 : Rat brain lysate
      Lane 4 : C6 (rat glial tumor glial cell line) whole cell lysate
      Lane 5 : Mouse cerebellum lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.

      Lanes 1 & 2: 15 seconds; Lanes 3 & 4: 3 minutes; Lane 5: 48 seconds.

      Negative control: NIH/3T3, C6 (lanes 2 & 4, PMID: 8370070, 9889325).

    References

    ab229915 has not yet been referenced specifically in any publications.

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