Recombinant Alexa Fluor® 488 Anti-Ki67 antibody [SP6] (ab281847)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Alexa Fluor® 488 Rabbit monoclonal [SP6] to Ki67
- Suitable for: IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Human
- Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm
Related conjugates and formulations
Overview
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Product name
Alexa Fluor® 488 Anti-Ki67 antibody [SP6]
See all Ki67 primary antibodies -
Description
Alexa Fluor® 488 Rabbit monoclonal [SP6] to Ki67 -
Host species
Rabbit -
Conjugation
Alexa Fluor® 488. Ex: 495nm, Em: 519nm -
Tested applications
Suitable for: IHC-P, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human and mouse colon tissues. ICC: HeLa cells. Flow Cyt (Intra): HeLa 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.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
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. Store In the Dark. -
Storage buffer
pH: 7.2
Preservative: 0.02% Sodium azide
Constituents: 30% Glycerol (glycerin, glycerine), 1% BSA, 68.98% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
SP6 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Dyes/Markers
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Immunohistochemistry reagents
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab281847 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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IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/50.
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Flow Cyt (Intra) |
1/5000.
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Notes |
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IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/50. |
Flow Cyt (Intra)
1/5000. |
Target
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Function
Required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly (PubMed:27362226). Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the chromosome surface (PubMed:27362226). Prevents chromosomes from collapsing into a single chromatin mass by forming a steric and electrostatic charge barrier: the protein has a high net electrical charge and acts as a surfactant, dispersing chromosomes and enabling independent chromosome motility (PubMed:27362226). Binds DNA, with a preference for supercoiled DNA and AT-rich DNA (PubMed:10878551). Does not contribute to the internal structure of mitotic chromosomes (By similarity). May play a role in chromatin organization (PubMed:24867636). It is however unclear whether it plays a direct role in chromatin organization or whether it is an indirect consequence of its function in maintaining mitotic chromosomes dispersed. -
Sequence similarities
Contains 1 FHA domain.
Contains 16 K167R repeats.
Contains 1 PP1-binding domain. -
Developmental stage
Expression occurs preferentially during late G1, S, G2 and M phases of the cell cycle, while in cells in G0 phase the antigen cannot be detected (at protein level) (PubMed:6206131). Present at highest level in G2 phase and during mitosis (at protein level). In interphase, forms fiber-like structures in fibrillarin-deficient regions surrounding nucleoli (PubMed:2674163, PubMed:8799815). -
Post-translational
modificationsPhosphorylated. Hyperphosphorylated in mitosis (PubMed:10502411, PubMed:10653604). Hyperphosphorylated form does not bind DNA. -
Cellular localization
Chromosome. Nucleus. Nucleus, nucleolus. Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the mitotic chromosome surface (PubMed:27362226). Associates with satellite DNA in G1 phase (PubMed:9510506). Binds tightly to chromatin in interphase, chromatin-binding decreases in mitosis when it associates with the surface of the condensed chromosomes (PubMed:15896774, PubMed:22002106). Predominantly localized in the G1 phase in the perinucleolar region, in the later phases it is also detected throughout the nuclear interior, being predominantly localized in the nuclear matrix (PubMed:22002106). - Information by UniProt
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Database links
- Entrez Gene: 4288 Human
- Entrez Gene: 17345 Mouse
- Omim: 176741 Human
- SwissProt: P46013 Human
- SwissProt: E9PVX6 Mouse
- SwissProt: Q61769 Mouse
- Unigene: 689823 Human
- Unigene: 80976 Human
see all -
Alternative names
- Antigen identified by monoclonal antibody Ki 67 antibody
- Antigen identified by monoclonal antibody Ki-67 antibody
- Antigen KI-67 antibody
see all
Images
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Ki67 with ab281847.
Positive staining on human colon. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab281847 at 1/50 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling Ki67 with ab281847 at 1/50 dilution (10.0 µg/ml) (Green).
Confocal image showing nucleolar staining in HeLa cells. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma cell) cells labelling Ki67 with ab281847 at 1/5000 dilution (0.01µg) (Right) compared with a Rabbit monoclonal isotype control - Alexa Fluor® 488 (Left) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Cells were co-stained with DRAQ5 to differentiate cell cycle phase.
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Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Ki67 with ab281847.
Positive staining on mouse colon. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab281847 at 1/50 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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 (1)
ab281847 has been referenced in 1 publication.
- Zhang Y et al. Nanohydroxyapatite Hydrogel Can Promote the Proliferation and Migration of Chondrocytes and Better Repair Talar Articular Cartilage. Comput Math Methods Med 2022:8388473 (2022). PubMed: 35664641