Antibody data
- Antibody Data
- Antigen structure
- References [8]
- Comments [0]
- Validations
- Immunocytochemistry [1]
- Other assay [5]
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- Product number
- 42-5699-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Ki-67 Monoclonal Antibody (20Raj1), eFluor™ 615, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The monoclonal antibody 20Raj1 recognizes the human Ki-67 protein. Two isoforms of Ki-67 exist, a 345 and 395 kDa form that are expressed in dividing cells. Ki-67 is expressed in all cell types and is detectable during active phases of the cell cycle (G1, S, G2, and mitosis) but is absent from resting cells (G0). During interphase, Ki-67 expression is localized to the nucleus but redistributes to the chromosomes during mitosis and has specifically been found to associate with heterochromatin-bound proteins such as chromobox protein homolog 3 (CBX3). In studies of tumor cells, Ki-67 expression has been used as a marker for determining the fraction of proliferating cells within a given population of tumor cells. This monoclonal antibody 20Raj1 recognizes canine Ki-67. Applications Reported: This 20Raj1 antibody has been reported for use in immunocytochemical and immunohistochemical staining of paraffin-embedded tissue (IHC-P). Applications Tested: This 20Raj1 antibody has been tested by immunocytochemistry on formaldehyde-fixed and permeabilized HeLa cells, and immunohistochemistry on FFPE human skin using low pH antigen retrieval. For immunocytochemistry and immunohistochemistry, this can be used at less than or equal to 1 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Filter Recommendation: When using this eFluor® 615 antibody conjugate, we recommend a filter that will capture the 615 emission wavelength. A standard Alexa Fluor® 594 filter is acceptable. Excitation: 595 nm; Emission: 615 nm. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human, Canine
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 20Raj1
- Vial size
- 100 µg
- Concentration
- 0.2 mg/mL
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Core Transcription Factors, MicroRNAs, and Small Molecules Drive Transdifferentiation of Human Fibroblasts Towards The Cardiac Cell Lineage.
S100A6 Regulates Endothelial Cell Cycle Progression by Attenuating Antiproliferative Signal Transducers and Activators of Transcription 1 Signaling.
Biliary tree stem cells, precursors to pancreatic committed progenitors: evidence for possible life-long pancreatic organogenesis.
Ki-Mcm6, a new monoclonal antibody specific to Mcm6: comparison of the distribution profile of Mcm6 and the Ki-67 antigen.
Detection of growth fraction in tumors by Ki67 monoclonal antibody in cytologic smears: a prospective study of 40 cases.
The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins.
Ki-67 immunostaining in uveal melanoma. The effect of pre-enucleation radiotherapy.
Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67.
Christoforou N, Chakraborty S, Kirkton RD, Adler AF, Addis RC, Leong KW
Scientific reports 2017 Jan 10;7:40285
Scientific reports 2017 Jan 10;7:40285
S100A6 Regulates Endothelial Cell Cycle Progression by Attenuating Antiproliferative Signal Transducers and Activators of Transcription 1 Signaling.
Lerchenmüller C, Heißenberg J, Damilano F, Bezzeridis VJ, Krämer I, Bochaton-Piallat ML, Hirschberg K, Busch M, Katus HA, Peppel K, Rosenzweig A, Busch H, Boerries M, Most P
Arteriosclerosis, thrombosis, and vascular biology 2016 Sep;36(9):1854-67
Arteriosclerosis, thrombosis, and vascular biology 2016 Sep;36(9):1854-67
Biliary tree stem cells, precursors to pancreatic committed progenitors: evidence for possible life-long pancreatic organogenesis.
Wang Y, Lanzoni G, Carpino G, Cui CB, Dominguez-Bendala J, Wauthier E, Cardinale V, Oikawa T, Pileggi A, Gerber D, Furth ME, Alvaro D, Gaudio E, Inverardi L, Reid LM
Stem cells (Dayton, Ohio) 2013 Sep;31(9):1966-79
Stem cells (Dayton, Ohio) 2013 Sep;31(9):1966-79
Ki-Mcm6, a new monoclonal antibody specific to Mcm6: comparison of the distribution profile of Mcm6 and the Ki-67 antigen.
Heidebrecht HJ, Buck F, Endl E, Kruse ML, Adam-Klages S, Andersen K, Frahm SO, Schulte C, Wacker HH, Parwaresch R
Laboratory investigation; a journal of technical methods and pathology 2001 Aug;81(8):1163-5
Laboratory investigation; a journal of technical methods and pathology 2001 Aug;81(8):1163-5
Detection of growth fraction in tumors by Ki67 monoclonal antibody in cytologic smears: a prospective study of 40 cases.
Rishi M, Schwarting R, Kovatich AJ, Ehya H
Diagnostic cytopathology 1993;9(1):52-6; dicussion 57-8
Diagnostic cytopathology 1993;9(1):52-6; dicussion 57-8
The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins.
Schlüter C, Duchrow M, Wohlenberg C, Becker MH, Key G, Flad HD, Gerdes J
The Journal of cell biology 1993 Nov;123(3):513-22
The Journal of cell biology 1993 Nov;123(3):513-22
Ki-67 immunostaining in uveal melanoma. The effect of pre-enucleation radiotherapy.
Mooy CM, de Jong PT, Van der Kwast TH, Mulder PG, Jager MJ, Ruiter DJ
Ophthalmology 1990 Oct;97(10):1275-80
Ophthalmology 1990 Oct;97(10):1275-80
Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67.
Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H
Journal of immunology (Baltimore, Md. : 1950) 1984 Oct;133(4):1710-5
Journal of immunology (Baltimore, Md. : 1950) 1984 Oct;133(4):1710-5
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry of fixed and permeabilized HeLa cells using 1 µg/mL of Anti-Human Ki-67 eFluor® 615.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 2 Immunofluorescence characterization of transdifferentiated iCM (cardiac TF and microRNA). ( A ) Expression of ACTN2 and TNNT2 or ( B ) Expression of TNNT2 and Ki67 in HDF transdifferentiated for 2 weeks using induced expression of GATA4, TBX5, MEF2C, MYOCD, NKX2-5 and transfection with hsa-miR-1 and hsa-miR-133a. (Controls: M2rtTA only, microRNA only, cardiac TF only). ( C ) ACTN2. Panels on the right show varying degrees of cytoskeletal organization of ACTN2. ( D ) TNNT2. Panels on the right show varying degrees of cytoskeletal organization of TNNT2. ( E ) ACTN2 and GJA1. ( F ) TNNT2 and Ki67 expression in iCM. ( G ) VIM and MYH6/7. ( H ) TAGLN. ( I ) SMA. ( J ) MYH11. ( K ) ACTN2 expression in iCM 4 weeks following initiation of transdifferentiation. Panels on the right show varying degrees of cytoskeletal organization of ACTN2. ( L ) TNNT2 expression in iCM 4 weeks following initiation of transdifferentiation. Panels on the right show varying degrees of cytoskeletal organization of TNNT2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 6 Determining the effect of small molecule inhibitors on transdifferentiation efficiency. ( A ) Immunofluorescence of transdifferentiated cells (ACTN2/TNNT2) during exposure to small molecule inhibitors: Janus protein tyrosine kinase 1 (JAK1i), Sodium Butyrate (HDACi), SB431542 (TGFbetai), CHIR99021 (GSK3i). ( B ) Number of nuclei, ( C ) Number of ACTN2 + cells, and ( D ) Number of TNNT2 + cells (per mm 2 ). ( E ) Number of Ki67 + nuclei normalized to the total number of nuclei. ( F ) Number of ACTN2 + cells normalized to the total number of nuclei. ( G ) Number of TNNT2 + cells normalized to the total number of nuclei. Experiment performed in triplicate. 4 images were analyzed for each experiment. Error bar represents calculated standard deviation. Significant difference between two values was calculated using t-test (two-tailed distribution, two sample unequal variance).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 7 Determining the effect of protein ligands on transdifferentiation efficiency. ( A ) Immunofluorescence of transdifferentiated cells (ACTN2/TNNT2) during exposure to protein ligands: IGF1, EGF, NRG1, SDF1A. ( B ) Number of nuclei, ( C ) Number of ACTN2 + cells, and ( D ) Number of TNNT2 + cells (per mm 2 ). ( E ) Number of Ki67 + nuclei normalized to the total number of nuclei. ( F ) Number of ACTN2 + cells normalized to the total number of nuclei. ( G ) Number of TNNT2 + cells normalized to the total number of nuclei. Experiment performed in triplicate. 4 images were analyzed for each experiment. Error bar represents calculated standard deviation. Significant difference between two values was calculated using t-test (two-tailed distribution, two sample unequal variance).