Overview

  • Product name

    Anti-RCN1/RCN antibody [EPR19193]
    See all RCN1/RCN primary antibodies
  • Description

    Rabbit monoclonal [EPR19193] to RCN1/RCN
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse RCN1/RCN aa 50 to the C-terminus. The exact sequence is proprietary.
    Database link: Q05186

  • Positive control

    • WB: Human fetal liver, heart, kidney and spleen lysates; HeLa, HEK-293, U-87 MG, C6, SH-SY5Y, Neuro-2a, RAW 264.7, PC-12 and NIH/3T3 cell lysates; E14 mouse and rat embryo lysates; Rat cerebral cortex, brain, heart and spleen lysates; Mouse brain, heart and spleen lysates. IHC-P: Human cerebrum and thyroid cancer tissues; Mouse and rat cerebral cortex tissues. IHC-Fr: Mouse hippocampus tissue. ICC/IF: SH-SY5Y, Neuro-2a and PC-12 cells. Flow Cyt: SH-SY5Y and Neuro-2a cells. IP: NIH/3T3 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

    EPR19193
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab184441 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 1/2000. Detects a band of approximately 44, 46 kDa (predicted molecular weight: 38 kDa).
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/150.
ICC/IF 1/500.
IHC-Fr 1/250.

Antigen retrieval: Heated citric acid solution (10mM citric acid PH 6.0 + 0.05% Tween-20)

IP 1/40.

Target

Images

  • All lanes : Anti-RCN1/RCN antibody [EPR19193] (ab184441) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human fetal spleen 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 1/10000 dilution

    Predicted band size: 38 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, 2 and 3: 30 seconds; Lane 4: 3 minutes.

  • All lanes : Anti-RCN1/RCN antibody [EPR19193] (ab184441) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
    Lane 4 : E14 mouse embryo lysate
    Lane 5 : E14 rat embryo lysate
    Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 7 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
    Lane 8 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
    Lane 9 : Rat cerebral cortex 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: 38 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 1-6: 8 seconds; Lane 7 and 8: 30 seconds; Lane 9: 3 minutes.

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

  • All lanes : Anti-RCN1/RCN antibody [EPR19193] (ab184441) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 9 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 10 : NIH/3T3 (Mouse embryonic fibroblast cell line) 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: 38 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 1-6: 3 minutes Lane 7 and 8: 30 seconds; Lane 9 and 10: 1 second.

  • Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling RCN1/RCN with ab184441 at 1/4000 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 thyroid cancer tissue labeling RCN1/RCN with ab184441 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on tumor cells of Human thyroid cancer 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 ab184441 at 1/4000 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 cerebral cortex tissue labeling RCN1/RCN with ab184441 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on neurons of rat 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 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus tissue labeling RCN1/RCN with ab184441 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The result showed cytoplasmic staining on mouse hippocampus. The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 100% methanol-fixed SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling RCN1/RCN with ab184441 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 counter stain 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: ab184441 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 100% methanol-fixed Neuro-2a (Mouse neuroblastoma cell line) cells labeling RCN1/RCN with ab184441 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 counter stain 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: ab184441 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 100% methanol-fixed PC-12 (Rat adrenal gland pheochromocytoma cell line) cells labeling RCN1/RCN with ab184441 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 PC-12 cell line. The nuclear counter stain 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: ab184441 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 Neuro-2a (Mouse neuroblastoma cell line) cells labeling RCN1/RCN with ab184441 at 1/150 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.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling RCN1/RCN with ab184441 at 1/150 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.

  • RCN1/RCN was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab184441 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184441 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: NIH/3T3 whole cell lysate, 10µg (Input).

    Lane 2: ab184441 IP in NIH/3T3 whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab184441 in NIH/3T3 whole cell lysate.

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

    Exposure time: 1 second.

References

This product has been referenced in:

  • Chen X  et al. Overexpression of RCN1 correlates with poor prognosis and progression in non-small cell lung cancer. Hum Pathol 83:140-148 (2019). Read more (PubMed: 30172915) »
See 1 Publication for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab184441.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up