Recombinant
RabMAb

Recombinant Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)

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Overview

  • Product name

    Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free
    See all Calbindin primary antibodies

  • Description

    Rabbit monoclonal [EPR22698-236] to Calbindin - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, 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

    • 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

    Ab255691 is the carrier-free version of ab229915. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab255691 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab255691 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 at an assay dependent concentration. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
IP Use at an assay dependent concentration.
  • 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

      • Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)
        Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)

        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).

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229915).

      • Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)
        Immunohistochemistry (Frozen sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)

        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).

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229915).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)

        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

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229915).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)

        Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling Calbindin with ab229915 at 1/4000 dilution, followed by 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

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229915).

      • Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)
        Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229915).

      • Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)
        Immunoprecipitation - Anti-Calbindin antibody [EPR22698-236] - BSA and Azide free (ab255691)

        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.

        This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229915).

      References

      ab255691 has not yet been referenced specifically in any publications.

      Publishing research using ab255691? Please let us know so that we can cite the reference in this datasheet.

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