Invitrogen Ki-67 Monoclonal An

Invitrogen Ki-67 Monoclonal Antibody (SolA15), Biotin 应用：免疫印迹 (WB)、免疫组化 (IHC)、免疫组化（石蜡） (IHC (P))、免疫组化（冰冻） (IHC (F))、免疫细胞化学 (ICC/IF)、流式细胞分析 (Flow)等 

产品详细信息

Description: The monoclonal antibody SolA15 recognizes mouse and rat Ki-67, a 300 kDa nuclear protein. Ki-67 is present during all active phases of the cell cycle (G1, S, G2, and mitosis), but is absent from resting cells (G0). Ki-67 is detected within the nucleus during interphase but redistributes to the chromosomes during mitosis. Ki-67 is used as a marker for determining the growth fraction of a given population of cells. In studies of tumor cells, the "Ki-67 labeling index" refers to the number of Ki-67 positive cells within the population and this is used to predict outcome of particular cancer types. Ki-67 has been shown to interact with the DNA-bound protein chromobox protein homolog 3 (CBX3) (heterochromatin).

The SolA15 antibody also recognizes human, non-human primate and canine Ki-67.

Applications Reported: This SolA15 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, immunohistochemical staining of frozen tissue sections, immunohistochemical staining of formalin-fixed paraffin embedded tissue sections, microscopy, and immunocytochemistry.

Applications Tested: This SolA15 antibody has been tested immunocytochemistry of fixed and permeabilized C2C12 cells and can be used at less than or equal to 5 µg/mL or intracellular staining and flow cytometric analysis of stimulated mouse esplenocytes cells using the Foxp3/Transcription Factor Buffer Set (Product # 00-5523-00) and protocol. Please see BestProtocols® Section (Staining intracellular Antigens for Flow Cytometry) for staining protocol (refer to Protocol B: One-step protocol for intracellular (nuclear) proteins). This can be used at less than or equal to 0.125 µg per test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Filtration: 0.2 µm post-manufacturing filtered.

靶标信息

Ki-67 is a nuclear protein that is expressed during various stages in the cell cycle, particularly during late G1, S, G2, and M phases. The protein has a forkhead associated domain (FHA) through which it associates with euchromatin at the perichromosomal layer, the centromeric heterochromatin, and the nucleolus. Ki-67 is shown to have a cell cycle dependent topographical distribution with perinucleolar expression at G1, expression in the nuclear matrix at G2, and expression on the chromosomes during M phase. Ki-67 is commonly used as a proliferation marker because it is not detected in G0 cells, but increases steadily from G1 through mitosis. Ki-67 antibodies are useful in establishing the cell growing fraction in neoplasms. In neoplastic tissues, the prognostic value is comparable to the tritiated thymidine-labelling index. The correlation between low Ki-67 index and histologically low-grade tumors is strong. Ki-67 is routinely used as a neuronal marker of cell cycling and proliferation.

应用案例

Ki-67 Antibody (13-5698-82) in WB

eLife 2016 - Figure 3--figure supplement 9. Ki-67-mutant NIH-3T3 cells proliferate normally. ( A ) Left, analysis of the indicated protein levels by Western blotting in NIH 3T3 WT clone 4 (W4) and Ki-67 mutant clones 14, 21 and 38. LC, loading control. ( B ) Left, growth curves of NIH 3T3 WT clone W4 and Ki-67 mutant clones 14, 21 and 38. NIH 3T3 WT and mutant cells were counted every day for 4 days. Right, cell cycle distribution of the WT clone W4 and 14, 21, 38 Ki67 mutant clones as analysed by FACS. ( C ) qRT-PCR analysis of Ki-67 mRNA in NIH 3T3 WT clone W4 and NIH Ki-67 mutant clones 14, 21 and 38. Normalized by mRNA expression of B2m (beta-2-microglobulin).

 

Ki-67 Antibody (13-5698-82) in ICC/IF

Neural regeneration research 2023 - Figure 5 PLO has no significant effects on OPC proliferation and does not interfere with the proliferative effect of FN. (A) Representative immunofluorescence images of OPCs stained for NG2 (red, stained with Alexa Fluor555, an OPC marker), Ki67 (green, stained with Alexa Fluor488, a proliferation marker), and DAPI (blue, staining for nuclei), cultured for 2 days in proliferation medium (2 d PM) under different coating conditions. The proliferation rate of OPCs in the PLO + FN group was similar to that in the FN group, which was higher than that in the PDL or PLO groups. Scale bar: 100 um. (B) Quantitative analysis of A, showing the proportion of Ki67 and NG2 co-labeled cells to NG2 cells. (C) Representative western blots of CyclinD1 protein from oligodendrocytes cultured for 2 days in proliferation medium (2 d PM). (D) Quantification of the CyclinD1 protein expression level (normalized by the PDL group) in C. Data were collected from at least three independent western blots. Data are expressed as mean +- SD from three independent experiments. * P < 0.05, ** P < 0.01 (one-way analysis of variance followed by Tukey's multiple comparison test). DAPI: 4',6-Diamidino-2-phenylindole; FN: fibronectin; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MBP: myelin basic protein; NG2: nerve-glia antigen 2; Olig2: oligodendrocyte transcription factor 2; OPC: oligodendrocyte precursor cell; PDL: poly-D-lysine; PLO: poly-L-ornithine; PM: proliferation medium.

 

Ki-67 Antibody (13-5698-82) in WB, ICC/IF

eLife 2016 - Figure 4. Cell proliferation without Ki-67. ( A ) Schematic representation of strategy for TALEN-mediated generation of Mki67 null allele. ( B ) qRT-PCR analysis of Ki-67 mRNA levels in NIH-3T3 WT clone W4 and Ki-67-negative 60, 65, 99 clones. ( C ) Top: Western blot of Ki-67 and Cyclin A in NIH-3T3 WT clone W4 and Ki-67-negative mutant clones 60, 65, 99; LC, loading control; below, Ki-67 immunofluorescence; bar, 10 um. ( D ) Left, growth curves of WT and Ki-67 null cell lines 60, 65 and 99; right, cell cycle distribution analysed by flow cytometry. ( E ) Cell cycle length of WT clone W4 and Ki-67 null clones 60 and 65 as determined by time-lapse videomicroscopy. ( F ) Cells of clone 65 show altered chromosomal periphery in mitosis. The Ki-67 staining is deliberately overexposed to demonstrate absence of detectable Ki-67 in clone 65, even in metaphase. Bar, 5 um.  1. Generation of NIH-3T3 cells lacking Ki-67. Top, schematic representation of the wild type (WT), knock-out or eGFP knock-in Mki67 locus and the predicted insertion of tandem repeats of the eGFP insert upstream of Mki67 locus. Bottom, Southern-blot of two NIH-3T3 WT clones (WT, W4) and six NIH-3T3 Ki-67 mutant clones (60,63,65,82,99). Clones were digested with PstI or SphI and probed with 5' or 3' probe, respectively.