Recombinant
RabMAb

Recombinant Anti-RCN1/RCN antibody [EPR17163-117] (ab210404)

Overview

  • Product name

    Anti-RCN1/RCN antibody [EPR17163-117]
    See all RCN1/RCN primary antibodies
  • Description

    Rabbit monoclonal [EPR17163-117] to RCN1/RCN
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    within Human RCN1/RCN aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: Q15293

  • Positive control

    • WB: Human placenta and fetal kidney lysates; HEK-293, SH-SY5Y, HeLa, Neuro-2a, C6, PC-12 and NIH/3T3 whole cell lysates; E14 mouse embryo lysate; E14 rat embryo lysate; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart, kidney and spleen lysates. IHC-P: Human cerebrum and lung adenocarcinoma tissues; Mouse cerebral cortex tissue; Rat hippocampus tissue. ICC/IF: SH-SY5Y, Neuro-2a and NIH/3T3 cells. Flow Cyt: NIH/3T3 and SH-SY5Y cells. IP: 293T whole cell lysate.
  • General notes

     This product was previously labelled as RCN1

     

    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

  • 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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17163-117
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab210404 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 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 44, 46 kDa (predicted molecular weight: 39 kDa).
ICC/IF 1/500.
Flow Cyt 1/700.

Target

Images

  • All lanes : Anti-RCN1/RCN antibody [EPR17163-117] (ab210404) at 1/5000 dilution

    Lane 1 : Human placenta lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
    Lane 5 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 6 : E14 mouse embryo lysate
    Lane 7 : E14 rat embryo lysate
    Lane 8 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 39 kDa
    Observed band size: 44,46 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 8416973).

  • All lanes : Anti-RCN1/RCN antibody [EPR17163-117] (ab210404) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Rat kidney lysate
    Lane 8 : Rat spleen lysate
    Lane 9 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 10 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 11 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 39 kDa
    Observed band size: 44,46 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-8: 1 minute; Lane 9,10 and 11: 10 seconds.

  • Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on neurons of Human cerebrum is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Human lung adenocarcinoma tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on tumor cells of Human lung adenocarcinoma is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on neurons of mouse cerebral cortex is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labeling RCN1/RCN with ab210404 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasm staining on neurons of rat hippocampus is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling RCN1/RCN with ab210404 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on SH-SY5Y cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab210404 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (Mouse neuroblastoma cell line) cells labeling RCN1/RCN with ab210404 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on Neuro-2a cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab210404 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling RCN1/RCN with ab210404 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on NIH/3T3 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab210404 at 1/500 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling RCN1/RCN with ab210404 at 1/700 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

    Cells were permeabilised with 90% methanol (diluted with 1X PBS).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling RCN1/RCN with ab210404 at 1/700 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

    Cells were permeabilised with 90% methanol (diluted with 1X PBS).

  • RCN1/RCN was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab210404 at 1/50 dilution.

    Western blot was performed from the immunoprecipitate using ab210404 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: 293T whole cell lysate, 10µg (Input).

    Lane 2: ab210404 IP in 293T whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab210404 in 293T whole cell lysate.

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

    Exposure time: 1 second.

References

ab210404 has not yet been referenced specifically in any publications.

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