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|Antigen||Kinesin Heavy Chain Member 2A (KIF2A) Antibodies|
|Reactivity||Human, Mammalian, Mouse (Murine), Pig (Porcine), Rat (Rattus) Alternatives|
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP), Simple Western (SimWes), Western Blotting (WB)
|13 references available|
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|Immunogen||A recombinant segment of the N-terminal domain of human Kif2a. [UniProt# O00139]|
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|Alternative Name||Kif2a (KIF2A Antibody Abstract)|
|Background||Gene Symbol: KIF2A|
|Molecular Weight||Theoretical MW: 110 kDa|
|Pathways||Microtubule Dynamics, Ribonucleoprotein Complex Subunit Organization|
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|Application Notes||Western Blot 1:10000, Simple Western 1:1000, Immunocytochemistry/Immunofluorescence 1:10000, Immunoprecipitation 1:10-1:500This Kif2a antibody is useful for Western blot, Immunoprecipitation, and Immunocytochemistry/Immunofluorescence. A band at approx. 110 kDa can be detected by Western blot. In Simple Western only 10 - 15 μL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.|
The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.
Western blot Protocol for Kif2a antibody
Western Blot Protocol:
1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute rabbit anti-Kif2a primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.ICC/IF Protocol for Kif2a antibody Immunocytochemistry ProtocolCulture cells to appropriate density on suitable glass coverslips in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 5-10 minutes.
. Remove the formalin and add 0.5 % Triton-X 100 in TBS to permeabilize the cells. Incubate for 5-10 minutes.
. Remove the permeabilization buffer and add wash buffer (i.e. PBS or PBS with 0.1 % Tween-20). Be sure to not let the specimen dry out. Gently wash three times for 10 minutes.
. Alternatively, cells can be fixed with -20C methanol for 10 min at room temperature. Remove the methanol and rehydrate in PBS for 10 min before proceeding.
. To block nonspecific antibody binding incubate in 10 % normal goat serum for 1 hour at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature for 1 hour or at 4 degrees C overnight.
. Remove primary antibody and replace with wash buffer. Gently wash three times for 10 minutes.
. Add secondary antibody at the appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash buffer. Gently wash three times for 10 minutes.
. Nuclei can be staining with 4',6' diamino phenylindole (DAPI) at 0.1 µg/ml, or coverslips can be directly mounted in media containing DAPI.
. Cells can now be viewed with a fluorescence microscope.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow proper laboratory procedures for the disposal of formalin.
|Restrictions||For Research Use only|
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Buffer contains: No Preservative
|Handling Advice||Avoid freeze-thaw cycles|
|Storage||-80 °C,-20 °C|
|Storage Comment||Aliquot and store at -20°C or -80°C. Avoid freeze-thaw cycles.|
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|Product cited in:||
Park, Song, Kwon, Jang: "Ska1 cooperates with DDA3 for spindle dynamics and spindle attachment to kinetochore." in: Biochemical and biophysical research communications, Vol. 470, Issue 3, pp. 586-92, 2016 (Sample species: Human). Further details: Immunocytochemistry,Western Blotting,Immunofluorescence
Kwon, Park, Song, Jang: "DDA3 and Mdp3 modulate Kif2a recruitment onto the mitotic spindle to control minus-end spindle dynamics." in: Journal of cell science, Vol. 129, Issue 14, pp. 2719-25, 2016 Further details: Immunocytochemistry,Immunofluorescence
Uematsu, Okumura, Tonogai, Joo-Okumura, Alemayehu, Nishikimi, Fukui, Nakatsukasa, Kamura: "ASB7 regulates spindle dynamics and genome integrity by targeting DDA3 for proteasomal degradation." in: The Journal of cell biology, Vol. 215, Issue 1, pp. 95-106, 2016
Bendre, Rondelet, Hall, Schmidt, Lin, Brouhard, Bird: "GTSE1 tunes microtubule stability for chromosome alignment and segregation by inhibiting the microtubule depolymerase MCAK." in: The Journal of cell biology, Vol. 215, Issue 5, pp. 631-647, 2016 Method employed by authors: Immunohistochemistry (IHC) (Sample species: Mouse (Murine)).
Chen, Liu, Wang, Liu, Xu, Zhang, Li, Xu, Lin, He, Liao, Fu, Wang, Yang, Wang: "KIF2A regulates the spindle assembly and the metaphase I-anaphase I transition in mouse oocyte." in: Scientific reports, Vol. 6, pp. 39337, 2016 Method employed by authors: Immunofluorescence (fixed cells) (IF/ICC) (Sample species: Mouse (Murine)).
Yi, Ma, Liang, Zhang, Xu, Meng, Ouyang, Hou, Schatten, Sun, Quan: "Kif2a regulates spindle organization and cell cycle progression in meiotic oocytes." in: Scientific reports, Vol. 6, pp. 38574, 2016 Method employed by authors: Western Blotting (WB), Immunofluorescence (fixed cells) (IF/ICC) (Sample species: Mouse (Murine)).
Zaganjor, Weil, Gonzales, Minna, Cobb: "Ras transformation uncouples the kinesin-coordinated cellular nutrient response." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, Issue 29, pp. 10568-73, 2014 (Sample species: Human). Further details: Western Blotting
Ma, Tulu, Ferenz, Fagerstrom, Wilde, Wadsworth: "Poleward transport of TPX2 in the mammalian mitotic spindle requires dynein, Eg5, and microtubule flux." in: Molecular biology of the cell, Vol. 21, Issue 6, pp. 979-88, 2010 (Sample species: Pig (Porcine)). Further details: Immunocytochemistry,Immunofluorescence
Jang, Coppinger, Seki, Yates, Fang: "Plk1 and Aurora A regulate the depolymerase activity and the cellular localization of Kif2a." in: Journal of cell science, Vol. 122, Issue Pt 9, pp. 1334-41, 2009
Jang, Wong, Coppinger, Seki, Yates, Fang: "DDA3 recruits microtubule depolymerase Kif2a to spindle poles and controls spindle dynamics and mitotic chromosome movement." in: The Journal of cell biology, Vol. 181, Issue 2, pp. 255-67, 2008 (Sample species: Human). Further details: Immunocytochemistry,Western Blotting,Immunofluorescence
Ferenz, Wadsworth: "Prophase microtubule arrays undergo flux-like behavior in mammalian cells." in: Molecular biology of the cell, Vol. 18, Issue 10, pp. 3993-4002, 2007
Ganem, Compton: "The KinI kinesin Kif2a is required for bipolar spindle assembly through a functional relationship with MCAK." in: The Journal of cell biology, Vol. 166, Issue 4, pp. 473-8, 2004
Homma, Takei, Tanaka, Nakata, Terada, Kikkawa, Noda, Hirokawa: "Kinesin superfamily protein 2A (KIF2A) functions in suppression of collateral branch extension." in: Cell, Vol. 114, Issue 2, pp. 229-39, 2003
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