Recombinant Anti-ARPC5/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 ARPC5/p16 ARC
- Suitable for: ICC/IF, IP, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ARPC5/p16 ARC antibody [EP1551Y]
See all ARPC5/p16 ARC primary antibodies -
Description
Rabbit monoclonal [EP1551Y] to ARPC5/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
Suitable for: ICC/IF, IP, WB, IHC-Pmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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.
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
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
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 |
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ICC/IF |
1/50 - 1/100.
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IP |
1/20 - 1/70.
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WB | (5) |
1/1000 - 1/10000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).
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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. |
Notes |
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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. |
Target
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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
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Database links
- Entrez Gene: 10092 Human
- Entrez Gene: 67771 Mouse
- Entrez Gene: 360854 Rat
- Omim: 604227 Human
- SwissProt: O15511 Human
- SwissProt: Q9CPW4 Mouse
- SwissProt: Q4KLF8 Rat
- Unigene: 518609 Human
see all -
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
see all
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-ARPC5/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-ARPC5/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-ARPC5/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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab51243 has been referenced in 9 publications.
- Wang Y et al. 27-Hydroxycholesterol-Induced Dysregulation of Cholesterol Metabolism Impairs Learning and Memory Ability in ApoE ε4 Transgenic Mice. Int J Mol Sci 23:N/A (2022). PubMed: 36232940
- Indra I et al. Sensing Actin Dynamics through Adherens Junctions. Cell Rep 30:2820-2833.e3 (2020). PubMed: 32101754
- Wang Y et al. 27-Hydroxycholesterol Alters Synaptic Structural and Functional Plasticity in Hippocampal Neuronal Cultures. J Neuropathol Exp Neurol 78:238-247 (2019). PubMed: 30753597
- Choi J et al. Pimozide suppresses cancer cell migration and tumor metastasis through binding to ARPC2, a subunit of the Arp2/3 complex. Cancer Sci 110:3788-3801 (2019). PubMed: 31571309
- Moriya Y et al. Tumor suppressive microRNA-133a regulates novel molecular networks in lung squamous cell carcinoma. J Hum Genet 57:38-45 (2012). PubMed: 22089643
- Kinoshita T et al. Actin-related protein 2/3 complex subunit 5 (ARPC5) contributes to cell migration and invasion and is directly regulated by tumor-suppressive microRNA-133a in head and neck squamous cell carcinoma. Int J Oncol 40:1770-8 (2012). PubMed: 22378351
- Chang YF et al. Interaction between microRNAs and actin-associated protein Arpc5 regulates translational suppression during male germ cell differentiation. Proc Natl Acad Sci U S A 109:5750-5 (2012). ICC/IF ; Mouse . PubMed: 22447776
- Hubert T et al. Actin and Arp2/3 localize at the centrosome of interphase cells. Biochem Biophys Res Commun 404:153-8 (2011). ICC/IF ; Human . PubMed: 21108927
- Schmidt S et al. Kindlin-3-mediated signaling from multiple integrin classes is required for osteoclast-mediated bone resorption. J Cell Biol 192:883-97 (2011). ICC/IF ; Mouse . PubMed: 21357746