Anti-p16 ARC antibody [EP1551Y] (ab51243)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1551Y] to p16 ARC
- Suitable for: ICC/IF, IP, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-p16 ARC antibody [EP1551Y]
See all p16 ARC primary antibodies -
Description
Rabbit monoclonal [EP1551Y] to p16 ARC -
Host species
Rabbit -
Specificity
This antibody is specific to the p16 ARC protein, database link: O15511. There is no observed cross reactivity to the CDKN2A/p16INK4a protein.
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Tested Applications & Species
See all applications and species dataApplication Species ICC/IF MouseIHC-P HumanIP HumanWB MouseRatHuman -
Immunogen
Synthetic peptide within Human p16 ARC aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: O15511 -
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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1551Y -
Isotype
IgG -
Research areas
Images
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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.
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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.
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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).
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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.
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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 kDaBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
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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.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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