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RecombinantRabMAb

Recombinant Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

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Western blot - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Immunoprecipitation - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
  • Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25385-129] to CACNA1E - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), IP, WB, IHC-P
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free
    See all CACNA1E primary antibodies

  • Description

    Rabbit monoclonal [EPR25385-129] to CACNA1E - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, WB, IHC-Pmore details
    Unsuitable for: ICC or IHC-Fr

  • Species reactivity

    Reacts with: Mouse, Rat

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse brain lysate. Mouse and rat hippocampus lysate. IHC-P: Mouse striatum, rat hippocampus and rat brain tissues. Flow Cyt (Intra): Mouse and rat primary neuron cells. IP: Mouse brain.

  • General notes

    ab288772 is the carrier free version of ab288766.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab288772 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 262 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.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 262 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.
Application notes
Is unsuitable for ICC or IHC-Fr.

Target

  • Function

    Voltage-sensitive calcium channels (VSCC) mediate the entry of calcium ions into excitable cells and are also involved in a variety of calcium-dependent processes, including muscle contraction, hormone or neurotransmitter release, gene expression, cell motility, cell division and cell death. The isoform alpha-1E gives rise to R-type calcium currents. R-type calcium channels belong to the 'high-voltage activated' (HVA) group and are blocked by nickel, and partially by omega-agatoxin-IIIA (omega-Aga-IIIA). They are however insensitive to dihydropyridines (DHP), omega-conotoxin-GVIA (omega-CTx-GVIA), and omega-agatoxin-IVA (omega-Aga-IVA). Calcium channels containing alpha-1E subunit could be involved in the modulation of firing patterns of neurons which is important for information processing.

  • Tissue specificity

    Expressed in neuronal tissues and in kidney.

  • Sequence similarities

    Belongs to the calcium channel alpha-1 subunit (TC 1.A.1.11) family. CACNA1E subfamily.
    Contains 1 EF-hand domain.

  • Domain

    Each of the four internal repeats contains five hydrophobic transmembrane segments (S1, S2, S3, S5, S6) and one positively charged transmembrane segment (S4). S4 segments probably represent the voltage-sensor and are characterized by a series of positively charged amino acids at every third position.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • alpha-1 polypeptide antibody
      • BII antibody
      • Brain calcium channel II antibody
      • CAC 1E antibody
      • CAC1E antibody
      • CAC1E_HUMAN antibody
      • CACH 6 antibody
      • CACH6 antibody
      • CACNA1E antibody
      • Cacnl1 a6 antibody
      • CACNL1A6 antibody
      • Calcium channel antibody
      • Calcium channel, L type, alpha 1 polypeptide, isoform 6 antibody
      • calcium channel, R type antibody
      • Cav 2.3 antibody
      • Cchra 1 antibody
      • Cchra1 antibody
      • isoform 6 antibody
      • L type antibody
      • Voltage dependent R type calcium channel subunit alpha 1E antibody
      • Voltage gated calcium channel subunit alpha Cav2.3 antibody
      • Voltage-dependent R-type calcium channel subunit alpha-1E antibody
      • Voltage-gated calcium channel subunit alpha Cav2.3 antibody
      see all

    Images

    • Western blot - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Western blot - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      All lanes : Anti-CACNA1E antibody [EPR25385-129] (ab288766) at 1/1000 dilution

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse hippocampus tissue lysate
      Lane 3 : Mouse kidney tissue lysate
      Lane 4 : Rat hippocampus tissue lysate
      Lane 5 : Rat kidney tissue lysate

      Lysates/proteins at 40 µg per lane.

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

      Predicted band size: 262 kDa
      Observed band size: 257 kDa why is the actual band size different from the predicted?



      This data was developed using ab288766, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Lane 1: 103 seconds
      Lane 2-5: 70 seconds

      The molecular weight and expression pattern is consistent to what has been described in previous reports (PMID: 31704946, PMID: 30846929)

      Samples are non-boiled as boiling may cause protein aggregates. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded mouse striatum tissue labelling CACNA1E with ab288766 at 1/500 (0.956 µg/ml) followed by ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining in mouse striatum. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation. 

      Mouse primary neuron cell line was stained with primary ab288766 at 1/500 (Red). Rabbit monoclonal IgG (ab172730) (Black) was used as a isotype control at 1/2000 dilution. Cells were fixed in 4% paraformaldehyde and permeabilized with 90% methanol. 

      Secondary antibody used is Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at a 1/2000 dilution.

    • Immunoprecipitation - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Immunoprecipitation - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation.

      CACNA1E was immunoprecipitated from 0.35 mg mouse brain tissue lysate 10 µg with ab288766 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab288766 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse brain tissue lysate 10 µg

      Lane 2: ab288766 IP in mouse brain tissue lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab288766 in mouse brain tissue lysate

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

      Exposure time: 67 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labelling CACNA1E with ab288766 at 1/500 (0.956 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in rat hippocampus. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded rat brain tissue labelling CACNA1E with ab288766 at 1/500 (0.956 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in rat brain, and strong immunoreactivity was seen in the superficial layer of the cortex, hippocampus, and striatum. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded (A) HEK-293T transfected with a CACNA1E expression vector containing a his tag and (B) HEK-293T cells transfected with empty vector containing a his tag. Labelling CACNA1E with ab288766 at 1/500 (0.956 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T cells transfected with a CACNA1E expression vector containing a his tag. No staining on (B) HEK-293T cells transfected with empty vector containing a his tag. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labelling CACNA1E with ab288766 at 1/500 (0.956 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining in mouse kidney. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded rat kidney tissue labelling CACNA1E with ab288766 at 1/500 (0.956 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining in rat kidney. The section was incubated with ab288766 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Flow Cytometry (Intracellular) - Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

      This data was developed using ab288766, the same antibody clone in a different buffer formulation. 

      Rat primary neuron cell line was stained with primary ab288766 at 1/500 (Red). Rabbit monoclonal IgG (ab172730) (Black) was used as a isotype control at 1/2000. Cells were fixed in 4% paraformaldehyde and permeabilized with 90% methanol.

      Secondary antibody used is Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at a 1/2000 dilution.

    • Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)
      Anti-CACNA1E antibody [EPR25385-129] - BSA and Azide free (ab288772)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab288772? Please let us know so that we can cite the reference in this datasheet.

    ab288772 has not yet been referenced specifically in any publications.

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