Anti-Ki67 抗体 [37C7-12] (ab245113)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [37C7-12] to Ki67
- Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Ki67 antibody [37C7-12]
Ki67 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [37C7-12] to Ki67 -
由来種
Mouse -
アプリケーション
適用あり: Flow Cyt (Intra), ICC/IF, IHC-Pmore details
適用なし: WB -
種交差性
交差種: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- IHC-P: Human breast carcinoma and tonsil tissue. ICC/IF: Hap1-Ki67 cells Flow Cyt (intra): HAP1 cells.
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特記事項
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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
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 long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
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
37C7-12 -
アイソタイプ
IgG2a -
軽鎖の種類
kappa -
研究分野
関連製品
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Alternative Versions
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Related Products
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ab245113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) |
Use 0.02µg for 106 cells.
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ICC/IF | (1) |
Use a concentration of 1 - 2 µg/ml.
If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min. |
IHC-P |
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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特記事項 |
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Flow Cyt (Intra)
Use 0.02µg for 106 cells. |
ICC/IF
Use a concentration of 1 - 2 µg/ml. If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min. |
IHC-P
1/800. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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
- Omim: 176741 Human
- SwissProt: P46013 Human
- Unigene: 689823 Human
- Unigene: 80976 Human
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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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ab245113 staining Ki67 in Hap1-Ki67 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 1h. The cells were then incubated overnight at 4°C with ab245113 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Ki67 with ab245113 at 1/800 dilution, followed by a ready to use secondary. Nuclear staining on human breast carcinoma tissue is observed. The section was incubated with ab245113 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use secondary. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Ki67 with ab245113 at 1/100 dilution, followed by Goat Anti-Mouse IgG (Alexa Fluor® 488) (ab150113) secondary antibody at 1/1000 dilution (green). Confocal image showing nucleolus staining on HeLa cell line. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab179504 Anti-beta IV Tubulin antibody - Microtubule Marker at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 594) (ab150080) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab245113 at 1/50 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 594) (ab150080) secondary antibody at 1/1000 dilution.
-ve control 2: ab179504 at 1/200 dilution, followed by ab150113 AlexaFluor®488 Goat anti-Mouse secondary at 1/1000 dilution. -
Intracellular Intracellular Flow Cytometry overlay histogram showing wild-type HAP1 (green line) and MKI67 knockout HAP1 cells stained with ab245113 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab245113) (1x106 in 100 µl at 0.2 µg/ml) for 30 min at 22°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 22°C.
Isotype control antibody was mouse IgG2a&kappa (ab18413) used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line MKI67 knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Ki67 with ab245113 at 1/800 dilution, followed by a ready to use secondary. Nuclear staining on human tonsil tissue is observed. The section was incubated with ab245113 for 30 mins at RT. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use secondary. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. -
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HAP1 (human chronic myelogenous leukemia near-haploid cell line) cells labeling Ki67 with ab245113 at 1/100 dilution, followed by Goat Anti-Mouse IgG (Alexa Fluor® 488) (ab150113) secondary antibody at 1/1000 dilution (green). Confocal image showing nucleolus staining on HAP1 cell line. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab179504 Anti-beta IV Tubulin antibody - Microtubule Marker at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 594) (ab150080) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab245113 at 1/50 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 594) (ab150080) secondary antibody at 1/1000 dilution.
-ve control 2: ab179504 at 1/200 dilution, followed by ab150113 AlexaFluor®488 Goat anti-Mouse secondary at 1/1000 dilution.
プロトコール
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.
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 (17)
ab245113 は 17 報の論文で使用されています。
- Park JY et al. A microphysiological model of human trophoblast invasion during implantation. Nat Commun 13:1252 (2022). PubMed: 35292627
- Zhang L et al. Targeting the lncRNA FGD5-AS1/miR-497-5p/PD-L1 Axis Inhibits Malignant Phenotypes in Colon Cancer (CC). Biomed Res Int 2022:1133332 (2022). PubMed: 35845947
- Jiang H et al. Pyruvate Kinase M2 Mediates Glycolysis in the Lymphatic Endothelial Cells and Promotes the Progression of Lymphatic Malformations. Am J Pathol 191:204-215 (2021). PubMed: 33130045
- Huang Y et al. A Novel, Personalized Drug-Screening System for Platinum-Resistant Ovarian Cancer Patients: A Preliminary Clinical Report. Cancer Manag Res 13:2849-2867 (2021). PubMed: 33833569
- Gu Y & Zhou Z Berberine inhibits the proliferation, invasion and migration of endometrial stromal cells by downregulating miR-429. Mol Med Rep 23:N/A (2021). PubMed: 33846796