Immunofluorescent analysis of Kinesin 5B (green) in HeLa cells. Cells fixed in 3.2% paraformaldehyde and permeabilized with 0.2% Triton X-100 for 30 minutes at 37°C were blocked with 3% BSA in PBS for 30 minutes at room temperature. Cells were probed with a Kinesin 5B polyclonal antibody (Product # PA1-643) at a dilution of 1:250 overnight at 4°C, washed with PBS containing 0.2% Triton X-100, and incubated with a fluorescently-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:2000 for 1 hour at room temperature. Images were taken on wide-field fluorescent microscope at 63X magnification. Data courtesy of the Innovators Program.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C I(376) D E Q F D K E K A N L E A F T V D K D I(396)|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||5 µg|
|Western Blot (WB)||1:4000-1:6000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA1-643 detects kinesin 5B from mouse and human samples. This antibody is specific for kinesin 5B and does not detect other kinesin isotypes.
PA1-643 has been successfully used in Western blot and immunofluorescent procedures. By Western blot, this antibody detects an ~110 kDa protein representing kinesin 5B protein.
The PA1-643 immunizing peptide corresponds to amino acid residues 376-396 from human kinesin 5B. This sequence is 95% and 90% conserved for mouse and rat, respectively. This peptide (Cat. # PEP-198) is available for use in neutralization and control experiments.
Kinesins are a superfamily of microtubule-associated motor proteins involved in a variety of cellular processes including membranous organelle transport and cell division. Kinesin has been found in a variety of organisms and cell types and is subject to spatial and temporal regulation. These proteins have a modular structure including a conserved motor domain of approximately 350 amino acids, which is responsible for microtubule binding and ATP hydrolysis. In addition to the motor domain, subfamily members share common domain organization, exhibit sequence similarity, motility properties, and cellular functions outside of the motor domain.
There are currently three known Kinesin 5 family members denoted as A, B, and C. Kinesin 5A and kinesin 5C appear to be exclusively neuronal, whereas kinesin 5B appears to be ubiquitous in its expression.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Conventional kinesin holoenzymes are composed of heavy and light chain homodimers.
PA1-643 was used in western blot to investigate the composition of the kinesin holoenzymes
|DeBoer SR,You Y,Szodorai A,Kaminska A,Pigino G,Nwabuisi E,Wang B,Estrada-Hernandez T,Kins S,Brady ST,Morfini G||Biochemistry (47:4535)||2008|