Recombinant Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1614Y] to Cytokeratin 15
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cytokeratin 15 antibody [EPR1614Y]
See all Cytokeratin 15 primary antibodies -
Description
Rabbit monoclonal [EPR1614Y] to Cytokeratin 15 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human Cytokeratin 15 aa 400-500 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: Mouse skin tissue lysates; A-431 cell lysate. IHC-P: Human squamous cervical carcinoma, normal skin, cervical carcinoma tissue ICC/IF: HeLa cells. Flow Cyt (intra): A-431 cells.
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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. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 5.60 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1614Y -
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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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab52816 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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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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ICC/IF
1/50 - 1/100. |
Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Tissue specificity
Expressed in a discontinuous manner in the basal cell layer of adult skin epidermis, but continuously in the basal layer of fetal skin epidermis and nail. Also expressed in the outer root sheath above the hair bulb in hair follicle (at protein level). Expressed homogeneously in all cell layers of the esophagus and exocervix, but detected in the basal cell layer only of oral mucosa, skin and in the basal plus the next two layers of the suprabasal epithelium of the palate. -
Sequence similarities
Belongs to the intermediate filament family. - Information by UniProt
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Database links
- Entrez Gene: 3866 Human
- Entrez Gene: 16665 Mouse
- Omim: 148030 Human
- SwissProt: P19012 Human
- SwissProt: Q61414 Mouse
- Unigene: 654570 Human
- Unigene: 440010 Mouse
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Alternative names
- AI528832 antibody
- CK 15 antibody
- CK-15 antibody
see all
Images
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All lanes : Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816) at 1/10000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : KRT15 knockout A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab52816 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab52816 was shown to react with Cytokeratin 15 in wild-type A431 cells in Western blot with loss of signal observed in KRT15 knockout cell line ab263922 (knockout cell lysate ab262484). Wild-type A431 and KRT15 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab52816 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
ab52816 at 1/100 dilution staining Cytokeratin 15 in human squamous cervical carcinoma by Immunohistochemistry, Paraffin embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816) at 1/100000 dilution + Mouse skin tissue lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 45 kDa
Observed band size: 45 kDaBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM /TBSTExposure time: 5.5 seconds
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Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816) at 1/10000 dilution + fetal thymus lysate at 10 µg
Secondary
goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDa -
ab52816 at 1/50 dilution staining Cytokeratin 15 in HeLa cells by Immunocytochemistry.
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Overlay histogram showing A431 cells stained with ab52816 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52816, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
Fluorescent immunohistochemical analysis of paraffin-embedded human normal skin tissue using ab52816. Green-CK15 red-PI
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 15 antibody [EPR1614Y] (ab52816)
Fluorescent immunohistochemical analysis of paraffin-embedded human cervical carcinoma tissue using ab52816. Green-CK15 red-PI.
Perform heat mediated antigen retrieval with citrate buffer pH 6 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 (40)
ab52816 has been referenced in 40 publications.
- Zhong P et al. Low KRT15 expression is associated with poor prognosis in patients with breast invasive carcinoma. Exp Ther Med 21:305 (2021). PubMed: 33717248
- Morgun EI et al. Expression of Protein Kinases RIPK-1 and RIPK-3 in Mouse and Human Hair Follicle. Dokl Biochem Biophys 494:252-255 (2020). PubMed: 33119828
- Polec A et al. PML Regulates the Epidermal Differentiation Complex and Skin Morphogenesis during Mouse Embryogenesis. Genes (Basel) 11:N/A (2020). PubMed: 32992884
- Moses RL et al. Novel epoxy-tiglianes stimulate skin keratinocyte wound healing responses and re-epithelialization via protein kinase C activation. Biochem Pharmacol 178:114048 (2020). PubMed: 32446889
- Mertsch S et al. Decellularized human corneal stromal cell sheet as a novel matrix for ocular surface reconstruction. J Tissue Eng Regen Med 14:1318-1332 (2020). PubMed: 32652796