Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [32/N-Cadherin] to N Cadherin
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-N Cadherin antibody [32/N-Cadherin]
    See all N Cadherin primary antibodies

  • Description

    Mouse monoclonal [32/N-Cadherin] to N Cadherin

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control

    • WB: 292T, HeLa, HepG2, Mouse brain, kidney, Rat heart, C6, PC-12 lysates. IP: HeLa 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.

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.
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    32/N-Cadherin

  • Isotype

    IgG1

Images

  • Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    All lanes : Anti-N Cadherin antibody [32/N-Cadherin] (ab280375) at 1/1000 dilution

    Lane 1 : Wild-type 293T (human embryonic kidney epithelial cell), whole cell lysate
    Lane 2 : CDH2 knockout 293T (human embryonic kidney epithelial cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution

    Predicted band size: 99 kDa
    Observed band size: 140 kDa
    why is the actual band size different from the predicted?



    Blocking and Diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    Lanes 1 - 2: Merged signal (red and green). Green - ab280375 observed at 130 kDa. Red - loading control ab181602(Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

    Lanes 1-2: ab280375 Anti-CDH2 antibody was shown to react with CDH2 in 293T cells in Western blot. Loss of signal was observed when CDH2 knockout sample was used. Wild-type and CDH2 knockout samples were subjected to SDS-PAGE.

    ab280375 and Anti-GAPDH antibody [EPR16891] (ab181602) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    All lanes : Anti-N Cadherin antibody [32/N-Cadherin] (ab280375) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 99 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?



    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    Exposure time 26 seconds

    Negative control: MCF7 (PMID: 9177902)

  • Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    All lanes : Anti-N Cadherin antibody [32/N-Cadherin] (ab280375) at 1/5000 dilution

    Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
    Lane 2 : 293T (human embryonic kidney epithelial cell), whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 99 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?



    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    Exposure time 15 seconds

    Lanes 1-2 used the fresh lysates to minimize protein degradation.

  • Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    Western blot - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    All lanes : Anti-N Cadherin antibody [32/N-Cadherin] (ab280375) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Rat heart lysate
    Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
    Lane 5 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

    Predicted band size: 99 kDa
    Observed band size: 140 kDa why is the actual band size different from the predicted?



    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    Exposure time Lanes 1-2: 15 seconds

    Lanes 3: 10 seconds

    Lanes 4-5: 26 seconds

  • Immunoprecipitation - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    Immunoprecipitation - Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)

    N Cadherin was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab280375 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280375 at 1/1000 dilution. HRP-conjuagted mouse IgG for IP (ab131368) was used at 1/1000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

    Lane 2: ab280375 IP in HeLa whole cell lysate

    Lane 3: Mouse monoclonal IgG1 (ab18443) instead of ab280375 in HeLa whole cell lysate

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

    Exposure time: 23 seconds

    The molecular weight observed is consistent with what has been described in the literature (PMID: 22553038)

  • Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)
    Anti-N Cadherin antibody [32/N-Cadherin] (ab280375)

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