Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3610] to Ki67
- Suitable for: WB, IHC-P, Flow Cyt, ICC
- Knockout validated
- Reacts with: Human
参阅全部 Ki67 一抗
描述兔单克隆抗体[EPR3610] to Ki67
经测试应用适用于: WB, IHC-P, Flow Cyt, ICCmore details
不与反应: Mouse, Rat
Synthetic peptide within Human Ki67 aa 1050-1150. The exact sequence is proprietary.
Database link: P46013-1
- WB: HeLa and ramos cell lysates. IHC-P: Human tonsil, colon, ovarian carcinoma, squamous cell carcinoma of cervix and colonic adenocarcinoma tissues. ICC: HeLa, HT-29 cells, HAP1 cells. Flow Cyt: Ramos cells.
This product is not suitable for xenograft experiments. For further information please contact our Customer Services team.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
解离常数（KD）KD = 1.24 x 10 -11 M Learn more about KD
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
纯度Immunogen affinity purified
- Anti-Ki67 antibody [EPR3610] (Alexa Fluor® 647) (ab196907)
- Anti-Ki67 antibody [EPR3610] (Alexa Fluor® 488) (ab197234)
- Anti-Ki67 antibody [EPR3610] - BSA and Azide free (ab209897)
- Anti-Ki67 antibody [EPR3610] (Alexa Fluor® 568) (ab211968)
- Anti-Ki67 antibody [EPR3610] (HRP) (ab212215)
- Anti-Ki67 antibody [EPR3610] (Alexa Fluor® 555) (ab215226)
- Anti-Ki67 antibody [EPR3610] (Alexa Fluor® 594) (ab216709)
Our Abpromise guarantee covers the use of ab92742 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000. Predicted molecular weight: 359 kDa.
For unpurified use at 1/500 - 1/1000.
|IHC-P||1/500 - 1/1000.
For unpurified use at 1/500 - 1/1000.
Antigen retrieval: Heat in citrate buffer pH 6 for 20-30 min or enzymatic (trypsin, proteinase K) necessary if fixed in PFA.
Positive Control: Hu tonsil tissue, Ms - tumour, embryonic skin tissue, Rt - esophagus, small intestine and liver tissue
|Flow Cyt||1/100 - 1/150.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC||Use a concentration of 1 µg/ml.
For nuclear proteins, fixing cells in 4% PFA (20 min, room temp) is recommended.
It is recommended to incubate cells with 0.1% Triton-X for 5 min to detect nuclear antigen.
Positive Control: HeLa and HAP1 cells / Rt cardiomyocytes
功能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.
序列相似性Contains 1 FHA domain.
Contains 16 K167R repeats.
Contains 1 PP1-binding domain.
发展阶段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).
翻译后修饰Phosphorylated. Hyperphosphorylated in mitosis (PubMed:10502411, PubMed:10653604). Hyperphosphorylated form does not bind DNA.
细胞定位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
- Antigen identified by monoclonal antibody Ki 67 antibody
- Antigen identified by monoclonal antibody Ki-67 antibody
- Antigen KI-67 antibody
Comparison between RNApII-S2P-/low cells and Ki-67- cells
a: Regulation of Ki-67 and RNApII-S2P during proliferation and quiescence in T98G glioblastoma cells. T98G cells were grown in culture medium containing 10% (v/v) fetal bovine serum (FBS), were induced to become quiescent by serum starvation in medium supplemented with 0.5% (v/v) FBS for 14 days, and then were re-stimulated by being split 1:5 into new medium containing 10% (v/v) FBS and cultured for 3 days. The cells were detached from dishes with trypsin-EDTA solution, fixed in 10% (v/v) neutral buffered formalin, and centrifuged. Paraffin sections of the pellet were cut, and expression of Ki-67 and RNApII-S2P was examined by single (brown; a1, a2, a4, a5, a7, a8) or double immunostaining (Ki-67, brown; RNApII-S2P, red; a3, a6, a9). Hematoxylin (blue) was used as a nuclear stain. Ki-67- RNApII-S2P-/low cells (blue cells in the double stained sections) emerged only in the quiescent condition (a6, arrows). Scale bar, 10 μm. b: Single-color immunostaining for Ki-67 (b1) and RNApII-S2P (b2) in serial sections of glioblastoma tissue. Ki-67- tumor cells were frequently found, whereas only a few RNApII-S2P-/low cells (arrows) were observed around necrotic area. N, necrotic area; V, blood vessels. Scale bars, 50 μm.
Ki67 detected using ab92742.
(From Figure S2 of Ishii et al)
ab92742 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 1h. The cells were then incubated with ab92742 at 1µg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled in blue with DAPI.
Anti-Ki67 antibody [EPR3610] (ab92742) at 1/5000 dilution (purified) + Ramos (Human Burkitt's lymphoma cell line) cell lysate at 20 µg
Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 359 kDa
Observed band size: 395 kDa why is the actual band size different from the predicted?
Blocking and dilution buffer: 5% NFDM/TBST.
Overlay histogram showing Ramos (Human Burkitt's lymphoma cell line) cells stained with unpurified ab92742 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (unpurified ab92742, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events was performed.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human squamous cell carcinoma of cervix tissue labeling Ki67 with purified ab92742 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Counterstained with hematoxylin.
Negative control using PBS instead of primary antibody (inset).
Chromogenic triple immunostaining for estrogen receptor (ER), progesterone receptor (PgR), and Ki-67 in breast cancer tissue to verify the triple immunostaining detection method.
Panel e: ER+ PgR- Ki-67- cells were stained red (short arrow), ER- PgR+ Ki-67- cells were stained blue (black arrowhead), ER+ PgR+ Ki-67- cells were stained purple (long arrow), and Ki-67+ cells were stained brown (white arrowhead). These colors are easily distinguishable. Scale bars, 25 μm.
Deparaffinized sections were pretreated for antigen retrieval by boiling in antigen retrieval solution, pH 9. Sections were incubated with rabbit monoclonal antibody against Ki67 ab92742 at a 1/1000 dilution. After the reaction with (HRP)-conjugated secondary antibodies color was developed with (DAB) and sections were counterstained with hematoxylin.
Immunocytochemistry analysis of HT-29 (Human colorectal adenocarcinoma cell line) cells labeling Ki67 with purified ab92742 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human normal colon tissue labeling Ki67 with unpurified ab92742. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic adenocarcinoma tissue labeling Ki67 with unpurified ab92742. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma cell line) cells lablling Ki67 with purified ab92742 at 1/150 (red). Cells were fixed with 2% paraformaldehyde. An FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labeling Ki67 with unpurified ab92742. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Anti-Ki67 antibody [EPR3610] (ab92742) at 1/500 dilution (unpurified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg
HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 359 kDa
Observed band size: 395 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling Ki67 with unpurified ab92742. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
ab92742 staining Ki67 in human adenocarcinoma cells by ICC (Immunocytochemistry).
Cells were fixed with paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS and blocked with 5% serum for 1 hour at 21°C. Samples were incubated with primary antibody (1/1000) for 12 hours at 4°C. A Cy3® conjugated donkey anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/200.
Immunocytochemistry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Ki67 with unpurified ab92742 at a dilution of 1/250.
ab92742 被引用在 133 文献中.
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