Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E254] to c-Jun - ChIP Grade
  • Suitable for: WB, IHC-P, IP, ChIP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-c-Jun antibody [E254] - ChIP Grade
    See all c-Jun primary antibodies

  • Description

    Rabbit monoclonal [E254] to c-Jun - ChIP Grade

  • Host species

    Rabbit

  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested Applications & Species

    Application Species
    ChIP
    Rat
    IHC-P
    Human
    IP
    Mouse
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human c-Jun aa 1-100 (N terminal). The exact sequence is proprietary.

  • Epitope

    ab32137 reacts with an epitope located in the N terminal region of c-Jun.

  • Positive control

    • WB: NIH 3T3 and HeLa cell lysate. IHC-P: Human lung carcinoma tissue IP: HEK-293 and NIH3T3 cells.

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    All lanes : Anti-c-Jun antibody [E254] - ChIP Grade (ab32137) at 1/200 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in 1% SDS Hot lysis method.
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates prepared in RIPA lysis method
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates prepared in 1% SDS Hot lysis method 15ug.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 36 kDa
    Observed band size: 39 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and diluting buffer: 5% NFDM/TBST

    For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling c-Jun with purified ab32137 at 1:100 dilution (1.2 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Western blot - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Anti-c-Jun antibody [E254] - ChIP Grade (ab32137) at 1/5000 dilution + PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 36 kDa
    Observed band size: 43 kDa why is the actual band size different from the predicted?

  • Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)

    ab32137 (purified) at 1/60 dilution (20 µg/mL) immunoprecipitating c-Jun in HEK-293 whole cell lysate.
    Lane 2 (+): HEK-293(Human embronic kidney epithelial cell) whole cell lysate 10 µg
    Lane 3 (-): ab32137 & HEK-293 whole cell lysateRabbit monoclonal IgG (ab172730) instead of ab32137 in HEK-293 whole cell lysate
    For western blotting, ab32137 at 1/500 dilution (2.29 µg/mL) and
    VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM /TBST .

  • ChIP - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    ChIP - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Chromatin was prepared from PC-12 (starve overnight) + NGF ( 50 ng/ml 2h) cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab32137 (blue), and 20µl of protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5µg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
  • Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Immunoprecipitation - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    c-Jun was immunoprecipitated using 0.5mg NIH3T3 whole cell extract, 5µg of Rabbit polyclonal to c-Jun and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, NIH3T3 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32137.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 45kDa; c-Jun
  • OI-RD Scanning - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    OI-RD Scanning - Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)
    Anti-c-Jun antibody [E254] - ChIP Grade (ab32137)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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