Overview

  • Product name
    Anti-p16 ARC antibody [EP1551Y]
    See all p16 ARC primary antibodies
  • Description
    Rabbit monoclonal [EP1551Y] to p16 ARC
  • Host species
    Rabbit
  • 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 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 IHC antigen retrieval protocols.

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

    • 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.

    • 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.

    • 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).

    • 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 1/5000 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
      All lanes : 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
      All lanes : 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 1/2000 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
      All lanes : 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:
    • Taniguchi K  et al. Depletion of gamma-glutamylcyclotransferase in cancer cells induces autophagy followed by cellular senescence. Am J Cancer Res 8:650-661 (2018). Read more (PubMed: 29736310) »
    • Chen HQ & Gao D Inhibitory effect of microRNA-154 targeting WHSC1 on cell proliferation of human skin squamous cell carcinoma through mediating the P53 signaling pathway. Int J Biochem Cell Biol 100:22-29 (2018). WB ; Human . Read more (PubMed: 29727714) »
    See all 36 Publications for this product

    Customer reviews and Q&As

    1-10 of 13 Abreviews or Q&A

    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

    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

    Abcam user community

    Verified customer

    Submitted Feb 27 2015

    Answer

    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.
    2.1.2. De-paraffinize/hydrate using the following series of washes: two Xylene washes (5 min each), followed by two 100% ethanol rinses (5 min each), followed by 95% ethanol, 70% ethanol, 50% ethanol, 30% ethanol, followed by H2O and a TBST wash for 5 min on a shaker.
    2.2. Antigen Retrieval
    2.2.1 Immerse slides into staining dish containing Antigen Retrieval Solution (citrate pH 6 was used)
    2.2.2 Place covered staining dish into the rice cooker. Add 120 mL dH2O and press “cook”.
    2.2.3 When “cook” is turned to “warm” (about 20–30 min), unplug the cooker and remove the staining dish to the bench top.
    2.2.4 Allow to cool down, without cover, for 20 min.
    2.3. Staining
    2.3.1. Wash slides with TBST for 5 min on a shaker.
    2.3.2. Inactivate endogenous peroxidase by covering tissue with 3% hydrogen peroxide for 10 min.
    2.3.3. Wash slides three times with TBST (3 min each on a shaker).
    2.3.4. Block slides with the blocking solution for 1 hour.
    2.3.5. Dilute primary antibody in the blocking buffer per recommendation on the data sheet.
    2.3.6. Apply primary antibody to each section 1/10 dilution and incubate overnight in the humidified chamber (4 ºC).
    2.3.7. Wash slides three times with TBST (3 min each on a shaker).
    2.3.8. Apply to each section secondary HRP-conjugated anti-rabbit antibody diluted in the blocking solution per manufacturer’s recommendation; incubate for 1 hour at room temperature.
    2.3.9. Wash slides three times with TBST (3 min each on a shaker).
    2.3.10. Add freshly prepared DAB substrate to the sections.
    2.3.11. Incubate tissue sections with the substrate at room temperature until suitable staining develops (generally 2–5 min).
    2.3.12. Rinse sections with water.
    2.3.13. Counterstain with Hematoxylin.
    2.3.14. Rinse sections with water.
    2.3.15. Dehydrate samples using two rinses with 100% Ethanol (20 dips per rinse) followed by two rinses with Xylene (30 dips per rinse).
    2.3.16. Mount coverslips on slides using Permount medium.

    Read More

    Answer

    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 do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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    Answer

    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 hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    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 am sorry we are regrettably not able to provide any testing discounts for data and images from these applications for this particular antibody.

    We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

    As an alternative, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

    To find out more about our Abreview system, please see the following link:

    https://www.abcam.com/abreviews

    I hope this information is helpful. Please do not hesitate to contact me again if you have any other questions.

    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

    Abcam user community

    Verified customer

    Submitted May 17 2012

    Question
    Answer

    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.orders@abcam.com or by calling 1-888-772-2226. I'm also pleased to let you know that Abcam has generous loyalty schemes and promotions which seek to reward our valued customers thanking them for choosing Abcam as a supplier. Abpoints are awarded for Abreviews and more information can be found on our website. www.abcam.com/abreviews Please do not hesitate to contact us should you have further questions.

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    Answer

    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. As requested, I have issued a free of charge replacement with the order number XXXX. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

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    Answer

    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 need any more advice or information.

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