Anti-Ki67 抗体 [SP6] (ab16667)
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
- Suitable for: Flow Cyt (Intra), IHC-P, WB, mIHC, ICC
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Ki67 antibody [SP6]
Ki67 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [SP6] to Ki67 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), IHC-P, WB, mIHC, ICCmore details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Common marmoset -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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エピトープ
C-terminus -
ポジティブ・コントロール
- WB: HeLa cell lysate. IHC-P: Human tonsil, rat and mouse spleen tissues. ICC: HeLa and HAP1 cells. Flow Cyt (intra): HAP1 cells.
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特記事項
ab16667 was switched from a hybridoma to recombinant production method on 24th October 2019.
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.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.1% Sodium azide
Constituents: 1% BSA, PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
SP6 -
アイソタイプ
IgG -
研究分野
関連製品
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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
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KO cell pellets
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Related Products
- Ki67 peptide (ab15581)
- Fix & Perm / Cell Fixation & Permeabilization Kit (Flow Cytometry) (ab185917)
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812)
- Anti-PCNA antibody [PC10] (ab29)
- Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker (ab6326)
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab16667の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
1/1000.
ab172730, Rabbit monoclonal isotype, is suitable for use as an isotype control with this antibody. |
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IHC-P | (62) |
1/200.
Antigen retrieval: Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). Primary antibody condition: primary antibody incubation overnight at +4°C is recommended. |
WB | (2) |
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa.
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mIHC |
Use at an assay dependent concentration.
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ICC | (5) |
1/250.
If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min. |
特記事項 |
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Flow Cyt (Intra)
1/1000. ab172730, Rabbit monoclonal isotype, is suitable for use as an isotype control with this antibody. |
IHC-P
1/200. Antigen retrieval: Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). Primary antibody condition: primary antibody incubation overnight at +4°C is recommended. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa. |
mIHC
Use at an assay dependent concentration. |
ICC
1/250. If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min. |
ターゲット情報
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機能
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. -
配列類似性
Contains 1 FHA domain.
Contains 16 K167R repeats.
Contains 1 PP1-binding domain. -
発生段階
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). -
翻訳後修飾
Phosphorylated. Hyperphosphorylated in mitosis (PubMed:10502411, PubMed:10653604). Hyperphosphorylated form does not bind DNA. -
細胞内局在
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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参照データベース
- 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 -
別名
- Antigen identified by monoclonal antibody Ki 67 antibody
- Antigen identified by monoclonal antibody Ki-67 antibody
- Antigen KI-67 antibody
see all
画像
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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.
This image was generated from the hybridoma version.
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All lanes : Anti-Ki67 antibody [SP6] (ab16667)
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.
Predicted band size: 358 kDaLanes 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 cells. Loss of signal was observed when knockout cell line ab255407 (knockout cell lysate 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil sections labeling Ki67 with ab16667 at 1/200 (0.156 µg/mL).
Image A:
Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer)
Human tonsil tissue incubated with ab16667 overnight at +4°C.
Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).
Nuclear staining on human tonsil. The section was incubated with ab16667 overnight at +4°C.Image B:
Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection)
Human tonsil tissue incubated with ab16667 on a Leica Biosystems BOND® RX instrument.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution2) for 20 mins. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Rat spleen tissue. The section was pre-treated using heat mediated antigen retrieval with (citrate buffer, pH 6.0). The section was then incubated with ab16667, 1/200 (0.156 µg/mL) dilution and detected using ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on rat spleen. The section was incubated with ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Human tonsil tissue. The section was pre-treated using heat mediated antigen retrieval with ab93678 (citrate buffer, pH 6.0). The section was then incubated with ab16667, 1/200 (0.156 µg/ml) dilution and detected using ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on human tonsil is observed. The section was incubated with ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.
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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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Mouse spleen tissue. The section was pre-treated using heat mediated antigen retrieval with ab93678 (citrate buffer, pH 6.0). The section was then incubated with ab16667, 1/200 (0.156 µg/mL) dilution and detected using a ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on mouse spleen.The section was incubated with ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.
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Immunofluorescent analysis of 100% methanol-fixed, None permeabilized HeLa cells labelling Ki67 with ab16667 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in HeLa cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).
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Immunofluorescent analysis of 100% methanol-fixed, None permeabilized parental HAP1 cells labelling Ki67 with ab16667 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in parental HAP1cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized parental HAP1 (Wildtype control Human chronic myelogenous leukemia near-haploid cell line) cells labelling Ki67 with ab16667 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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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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Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)This image is courtesy of an anonymous abreview.
Immunocytochemistry/ Immunofluorescence analysis of human cardiac stem cells labeling Ki67 with ab16667 at 1/250 dilution. Cells were fixed in paraformaldehyde and permeabilized with Triton x-100, 0.01%. Cells were blocked in BSA for 1 hour at room temperature. A polyclonal chicken anti-rabbit Alex Fluor® 488 secondary antibody was used at 1/500 dilution.
This image was generated from the hybridoma version.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)Image Courtesy of Carl Hobbs, King College London, U.K.
ab16667 staining Ki67 in human testis by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% BSA for 10 minutes at 21ºC followed by incubation with the primary antibody for 2 hours at 1/100. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.
This image was generated from the hybridoma version.
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Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)This image is courtesy of an Abreview submitted by Peter Zentis
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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (1744)
ab16667 は 1744 報の論文で使用されています。
- Zhu X et al. HDAC1/2 Control Proliferation and Survival in Adult Epidermis and Pre?Basal Cell Carcinoma through p16 and p53. J Invest Dermatol 142:77-87.e10 (2022). PubMed: 34284046
- Yang W et al. Single-Cell Transcriptomic Analysis Reveals a Hepatic Stellate Cell-Activation Roadmap and Myofibroblast Origin During Liver Fibrosis in Mice. Hepatology 74:2774-2790 (2021). PubMed: 34089528
- Liu M et al. miR-140-5p inhibits the proliferation of multiple myeloma cells by targeting VEGFA. Mol Med Rep 23:N/A (2021). PubMed: 33200797
- Messal HA et al. Antigen retrieval and clearing for whole-organ immunofluorescence by FLASH. Nat Protoc 16:239-262 (2021). PubMed: 33247285
- Wang C et al. ER-a36 Promotes the Malignant Progression of Cervical Cancer Mediated by Estrogen via HMGA2. Front Oncol 11:712849 (2021). PubMed: 34336701