Validated using a knockout cell line
Recombinant
RabMAb

Anti-p16 ARC antibody [EP1551Y] (ab51243)

Overview

  • Product name
    Anti-p16 ARC antibody [EP1551Y]
    See all p16 ARC primary antibodies
  • Description
    Rabbit monoclonal [EP1551Y] to p16 ARC
  • Tested applications
    Suitable for: ICC/IF, IP, WB, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human p16 ARC aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control
    • WB: Human fetal, mouse and rat brain tissue lysates and HeLa, HepG2, SKBR-3 and MCF-7 cell lysates. IHC-P: Human uterus adenocarcinoma and spleen tissues. ICC/IF: Neuro-2a cells. IP: Human fetal brain tissue lysate.
  • General notes

    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.

    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. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EP1551Y
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab51243 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
ICC/IF 1/50 - 1/100.
IP 1/20 - 1/70.
WB 1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

We have obtained results that indicate IHC-P is unsuitable for mouse and rat species.  

  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Function
      Functions as component of the Arp2/3 complex which is involved in regulation of actin polymerization and together with an activating nucleation-promoting factor (NPF) mediates the formation of branched actin networks.
    • Sequence similarities
      Belongs to the ARPC5 family.
    • Cellular localization
      Cytoplasm > cytoskeleton. Cell projection.
    • Information by UniProt
    • Database links
    • Alternative names
      • Actin related protein 2/3 complex subunit 5 (16 kD) antibody
      • Actin related protein 2/3 complex subunit 5 antibody
      • Actin related protein 2/3 complex, subunit 5 16kDa antibody
      • Actin-related protein 2/3 complex subunit 5 antibody
      • ARC16 antibody
      • Arp2/3 complex 16 kDa subunit antibody
      • Arp2/3 protein complex subunit p16 antibody
      • ARPC 5 antibody
      • Arpc5 antibody
      • ARPC5_HUMAN antibody
      • dJ127C7.3 antibody
      • MGC88523 antibody
      • p16 Arc antibody
      • p16-ARC antibody
      • RP1 127C7.3 antibody
      see all

    Images



    • Predicted band size : 16 kDa

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: p16 ARC knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)

       

      Lanes 1 - 4: Merged signal (red and green). Green - ab51243 observed at 16 kDa. Red - loading control, ab8245, observed at 37 kDa.

       

      ab51243 was shown to specifically react with p16 ARC when p16 ARC knockout samples were used. Wild-type and p16 ARC knockout samples were subjected to SDS-PAGE. Ab51243 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/Immunofluorescence analysis of Neuro-2a cells labelling p16 ARC with purified ab51243 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    • All lanes : Anti-p16 ARC antibody [EP1551Y] (ab51243) at 1/5000 dilution (purified)

      Lane 1 : HeLa whole cell lysate
      Lane 2 : SKBR-3 whole cell lysate
      Lane 3 : MCF-7 whole cell lysate
      Lane 4 : HepG2 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 16 kDa
      Observed band size : 16 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • All lanes : Anti-p16 ARC antibody [EP1551Y] (ab51243) at 1/5000 dilution (purified)

      Lane 1 : Human fetal brain tissue lysate
      Lane 2 : Mouse brain tissue lysate
      Lane 3 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 16 kDa
      Observed band size : 16 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling p16 ARC with purified ab51243 at a dilution of 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • ab51243 (purified) at a dilution of 1/20 immunoprecipitating p16 ARC in human fetal brain tissue lysate.

      Lane 1 (input): Human fetal brain tissue lysate (10µg)

      Lane 2 (+): ab51243 + human fetal brain tissue lysate.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51243 in human fetal brain tissue lysate.

      For western blotting, a HRP-conjugated anti-rabbit specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-p16 ARC antibody [EP1551Y] (ab51243) at 1/2000 dilution (unpurified)

      Lane 1 : Human fetal brain tissue lysate
      Lane 2 : Mouse brain tissue lysate
      Lane 3 : Rat brain tissue lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 16 kDa

      Blocking buffer and concentration: 5% NFDM/TBST

      Diluting buffer and concentration: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human uterus adenocarcinoma tissue labelling p16 ARC with unpurified ab51243 at a dilution of 1/50.

    References

    This product has been referenced in:
    • Cheng XY  et al. AMP-activated protein kinase reduces inflammatory responses and cellular senescence in pulmonary emphysema. Oncotarget 8:22513-22523 (2017). WB ; Mouse . Read more (PubMed: 28186975) »
    • Yu T  et al. CBX7 is a glioma prognostic marker and induces G1/S arrest via the silencing of CCNE1. Oncotarget 8:26637-26647 (2017). WB . Read more (PubMed: 28460453) »

    See all 18 Publications for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain and Spleen)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Specification
    Brain and Spleen
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Ester Canto Puig

    Verified customer

    Submitted Jul 04 2016

    Application
    Western blot
    Loading amount
    25 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Mouse Tissue lysate - whole (mouse lung)
    Specification
    mouse lung
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 27 2015

    Thank you for contacing us. I have placed the protocol below, please let me know if you have any further questions.



    2. Protocol
    2.1. Deparaffinization/Rehydration
    2.1.1. Heat slides in an oven at 65 °C for 1 hour. Read More

    Thank you for contacting us.

    We do have control lysates for both recommended positive controls, human fetal brain tissue (ab29467) and human uterus adenocarcinoma tissue (ab44053).

    I hope this information is helpful to you. Please...

    Read More

    Thank you for contacting us.
    We have not specifically tested fat tissue in rat, however it is guaranteed to work in rat and should give a signal as long as the tissue expresses p16.

    I hope this information is helpful to you. Please do not ...

    Read More

    Thank you for your enquiry which has been forwarded to the scientific support team.

    I can confirm that ab51243 Anti-p16 ARC antibody [EP1551Y] has already been tested in IHC-P and WB. We also have images for these applications. Therefore, I ...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MCF7)
    Loading amount
    100000 cells
    Specification
    MCF7
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Username

    Abcam user community

    Verified customer

    Submitted May 17 2012

    Thank you for contacting us. While we do not currently offer a discount program specifically to XXXXXX, we do offer discountd off purchases of five products or more. Our customer service representative would be able to provide futher details at us.o...

    Read More

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused...

    Read More

    Thank you for contacting us. That is correct. I will process that for you as soon as you are able to send me the ooriginal order number or PO used to purchase this. I hope this information is helpful to you. Please do not hesitate to contact us if you ...

    Read More

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