Recombinant Anti-Ki67 antibody [SP6] - BSA free (ab231172)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP6] to Ki67 - BSA free
- Suitable for: mIHC, IHC-P, Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Ki67 antibody [SP6] - BSA free
See all Ki67 primary antibodies -
Description
Rabbit monoclonal [SP6] to Ki67 - BSA free -
Host species
Rabbit -
Tested applications
Suitable for: mIHC, IHC-P, Flow Cyt (Intra), WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Common marmoset -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Epitope
C-terminus -
Positive control
- WB: Ramos and HeLa cell lysates. IHC-P: Human tonsil and testis tissue. Rat esophagus, small intestine and liver tissue. Mouse embryonic skin tissue. Transgenic mouse spinal cord tissue. Human colon carcinoma. ICC/IF: HAP1 cells. Human cardiac stem cells. HEp-2 cells. Flow Cyt (intra): HAP1 cells.
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General notes
ab231172 is the carrier-free version of ab16667.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Affinity purified -
Clonality
Monoclonal -
Clone number
SP6 -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab231172 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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mIHC |
Use at an assay dependent concentration.
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IHC-P | (1) |
Use at an assay dependent concentration.
Use at an assay dependent concentration. Antigen retrieval: heat mediated antigen retrieval with sodium citrate buffer (pH 6) |
Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab172730, Rabbit monoclonal isotype, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa.
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ICC/IF |
Use at an assay dependent concentration.
Use at an assay dependent concentration. If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min. |
Notes |
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mIHC
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Use at an assay dependent concentration. Antigen retrieval: heat mediated antigen retrieval with sodium citrate buffer (pH 6) |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730, Rabbit monoclonal isotype, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa. |
ICC/IF
Use at an assay dependent concentration. Use at an assay dependent concentration. If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min. |
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
- Entrez Gene: 246042 Rat
- Omim: 176741 Human
- SwissProt: P46013 Human
- SwissProt: E9PVX6 Mouse
- SwissProt: Q61769 Mouse
- Unigene: 689823 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (ab231172)
This data was developed using the same antibody clone in a different buffer formulation (ab16667).
IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Human colon carcinoma tissue. The section incubated with ab16667 at 1/200 (0.145 μg/ml) dilution and ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (ab214880) was used as secondary antobody. Positive staining on human colon carcinoma. The section was incubated with ab16667 at 4°C overnight. The section was counterstained with haematoxylin.
Heat mediated antigen retrieval was performed using ab93678 (citrate buffer, pH 6.0).
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This data was developed using the same antibody clone in a different buffer formulation (ab16667).
Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).
Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.
This image was generated from the hybridoma version.
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All lanes : Anti-Ki67 antibody [SP6] (ab16667) at 1/100 dilution
Lane 1 : Ramos cell lysate
Lane 2 : Wild-type HeLa cell lysate
Lane 3 : MKI67 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 358 kDa
Observed band size: 359 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab16667).
Lanes 1-3: Merged signal (red and green). Green - ab16667 observed at 359 kDa. Red - loading control, ab130007 observed at 125 kDa.
ab16667 was shown to react with Ki67 in wild-type HeLa. Loss of signal was observed when knockout sample ab263762 was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. ab16667 and Anti-Vinculin antibody VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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This data was developed using the same antibody clone in a different buffer formulation (ab16667).
ab16667 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol for 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab16667 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (ab231172)
This data was developed using the same antibody clone in a different buffer formulation (ab16667).
IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BONDTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20mins. The section was then incubated with ab16667, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research CentreThis image was generated from the hybridoma version.
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This data was developed using the same antibody clone in a different buffer formulation (ab16667).
Overlay histogram showing HAP1 wildtype (green line) and HAP1-MKI67 knockout cells (red line) stained with ab16667. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab16667, 1/1000) for 30 min at 22°C. The secondary antibody used wasGoat Anti-Rabbit IgG H&L (Alexa Fluor®488) preadsorbed (ab150081) secondary antibodyat 1/2000 dilution for 30 min at 22°C. A Rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-MKI67 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. This image was generated from the hybridoma version.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA free (ab231172)Image courtesy of Jing Ma by Abreview.
This data was developed using the same antibody clone in a different buffer formulation (ab16667).
ab16667 staining Ki67 in rat small intestine tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 10 mM citrate buffer pH 6.0. Samples were then blocked with 10% serum for 20 minutes at room temperature followed by incubation with the undiluted primary antibody for 30 minutes. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/2000 dilution.
This image was generated from the hybridoma version.
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Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA free (ab231172)This image is courtesy of an Abreview submitted by Peter Zentis
This data was developed using the same antibody clone in a different buffer formulation (ab16667).
ab16667 staining Ki67 - Proliferation Marker in human HEp-2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with Triton X-100 0.25% in PBS. Samples were incubated with primary antibody (1/50 in DPBS) for 1 hour at 21°C. An Atto488-conjugated Donkey anti-rabbit polyclonal (1/50) was used as the secondary antibody.
This image was generated from the hybridoma version.
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 (21)
ab231172 has been referenced in 21 publications.
- Wei L et al. CMTM6 knockdown prevents glioma progression by inactivating the mTOR pathway. Ann Transl Med 10:181 (2022). PubMed: 35280358
- Li Y et al. CircRNA hsa_circ_0018289 exerts an oncogenic role in cervical cancer progression through miR-1294/ICMT axis. J Clin Lab Anal 36:e24348 (2022). PubMed: 35312113
- Cao B et al. Remodelling of tumour microenvironment by microwave ablation potentiates immunotherapy of AXL-specific CAR T cells against non-small cell lung cancer. Nat Commun 13:6203 (2022). PubMed: 36261437
- Tan L & Chen Z miR-193a-5p Enhances the Radioresistance of Pancreatic Cancer Cells by Targeting ZFP57 and Activating the Wnt Pathway. J Oncol 2022:8071343 (2022). PubMed: 36276285
- Li J et al. LncRNA LINC00473 is involved in the progression of invasive pituitary adenoma by upregulating KMT5A via ceRNA-mediated miR-502-3p evasion. Cell Death Dis 12:580 (2021). PubMed: 34091587