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RecombinantRabMAb

Recombinant Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
  • Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25332-124] to DNAJC6
  • Suitable for: IHC-P, WB, IHC-Fr, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-DNAJC6 antibody [EPR25332-124]
    See all DNAJC6 primary antibodies

  • Description

    Rabbit monoclonal [EPR25332-124] to DNAJC6

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, IHC-Fr, Flow Cyt (Intra)more details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • Flow cyt: U-87 MG and K-562 cells. IHC-P: Human brain tissue; Mouse brain tissue, Rat brain tissue. IHC-Fr: Mouse cortex tissue; Rat cortex tissue.

  • General notes

    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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR25332-124

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab289711 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-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 110 kDa (predicted molecular weight: 99 kDa).
IHC-Fr
1/50.
Flow Cyt (Intra)
1/500.
Notes
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 110 kDa (predicted molecular weight: 99 kDa).
IHC-Fr
1/50.
Flow Cyt (Intra)
1/500.
Application notes
Is unsuitable for IP.

Target

  • Relevance

    DNAJC6 belongs to the evolutionarily conserved DNAJ/HSP40 family of proteins, which regulate molecular chaperone activity by stimulating ATPase activity. DNAJ proteins may have up to 3 distinct domains: a conserved 70-amino acid J domain, usually at the N terminus, a glycine/phenylalanine (G/F)-rich region, and a cysteine-rich domain containing 4 motifs resembling a zinc finger domain.

  • Database links

    • Alternative names

      • auxilin antibody
      • DJC6 antibody
      • DnaJ (Hsp40) homolog, subfamily B, member 6 antibody
      • DnaJ (Hsp40) homolog, subfamily C, member 6 antibody
      • DnaJ homolog subfamily C member 6 antibody
      • EC 3.1.3.48 antibody
      • KIAA0473 antibody
      • MGC129914 antibody
      • MGC129915 antibody
      • MGC48436 antibody
      • putative tyrosine-protein phosphatase auxilin antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of paraffin-embedded Human brain tissue labelling DNAJC6 with ab289711 at 1/4000(0.114ug/ml) dilution  followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab289711 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 (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cortex (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution (Green). Positive staining on rat cortex. The nuclear conterstain is DAPI (Blue).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution.

       

    • Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast cell line) cells labeling DNAJC6 with ab289711 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081), at 1/2000 dilution was used as the secondary antibody.

    • Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      All lanes : Anti-DNAJC6 antibody [EPR25332-124] (ab289711) at 1/1000 dilution

      Lane 1 : Human spinal cord tissue lysate
      Lane 2 : Human liver tissue lysate
      Lane 3 : Human skeletal muscle tissue lysate
      Lane 4 : Mouse cerebral cortex tissue lysate
      Lane 5 : Mouse brain tissue lysate
      Lane 6 : Mouse liver tissue lysate
      Lane 7 : Mouse skeletal muscle tissue lysate
      Lane 8 : Rat brain tissue lysate
      Lane 9 : Rat liver tissue lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 99 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST

      Negative control: Liver, Skeletal muscle (PMID: 23211418)
      The molecular weight observed is consistent with what has been described in the literature (PMID: 1973169)

      Exposure times:

      Lane 1-7: 7.75 seconds
      Lane 8-9: 37 seconds

    • Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cortex (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution (Green). Positive staining on mouse cortex. The nuclear conterstain is DAPI (Blue).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution.

       

    • Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      All lanes : Anti-DNAJC6 antibody [EPR25332-124] (ab289711) at 1/1000 dilution

      Lane 1 : U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate
      Lane 2 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 99 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?


      Exposure time: 48 seconds


      Fresh lysates were used in this WB.
      The molecular weight observed is consistent with what has been described in the literature (PMID: 1973169). 

      5% NFDM/TBST was used as a blocking and diluting buffer. 

    • Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling DNAJC6 with ab289711 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081), at 1/2000 dilution was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labelling DNAJC6 with ab289711 at 1/4000(0.114ug/ml) dilution  followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum. The section was incubated with ab289711 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-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of paraffin-embedded Rat brain tissue labelling DNAJC6 with ab289711 at 1/4000(0.114ug/ml) dilution  followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab289711 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 (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution (Green). Negative control (PMID: 23211418). No staining on rat liver. The nuclear conterstain is DAPI (Blue).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling DNAJC6 with ab289711 at 1/4000(0.114ug/ml) dilution  followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: no staining on human liver (PMID: 25446072). The section was incubated with ab289711 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 (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution (Green). Negative control (PMID: 23211418). No staining on mouse liver. The nuclear conterstain is DAPI (Blue).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody at 1/1000 dilution.

    • Anti-DNAJC6 antibody [EPR25332-124] (ab289711)
      Anti-DNAJC6 antibody [EPR25332-124] (ab289711)

    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 ab289711? Please let us know so that we can cite the reference in this datasheet.

    ab289711 has not yet been referenced specifically in any publications.

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