Immunofluorescence analysis of Kinesin 5A was done on 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Kinesin 5A Rabbit Polyclonal Antibody (PA1642) at 1ug/ml in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Human, Mouse|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C(1007) G Y E A E D Q A K L F P L H Q E T A A S(1027)|
|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 *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||0.5-2 µg/ml|
* 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 3 publications below|
PA1-642 detects kinesin 5A from mouse, human, and bovine samples. This antibody is specific for kinesin 5A and does not detect other kinesin isotypes.
PA1-642 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~110 kDa protein representing kinesin 5A from mouse neuronal cell lysate and human retinal extracts.
The PA1-642 immunizing peptide corresponds to amino acid residues 1007-1027 from mouse kinesin 5A. This sequence is completely conserved in human. This peptide (Cat. # PEP-197) 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.
Neurochemical profile of dementia pugilistica.
PA1-642 was used in western blot to study the biochemical/molecular changes associated with dementia pugilistica
|Kokjohn TA,Maarouf CL,Daugs ID,Hunter JM,Whiteside CM,Malek-Ahmadi M,Rodriguez E,Kalback W,Jacobson SA,Sabbagh MN,Beach TG,Roher AE||Journal of neurotrauma (30:981)||2013|
APP anterograde transport requires Rab3A GTPase activity for assembly of the transport vesicle.
PA1-642 was used in western blot to investigate the role of the Rab3A GTPase during the APP anterograde transport.
|Szodorai A,Kuan YH,Hunzelmann S,Engel U,Sakane A,Sasaki T,Takai Y,Kirsch J,Müller U,Beyreuther K,Brady S,Morfini G,Kins S||The Journal of neuroscience : the official journal of the Society for Neuroscience (29:14534)||2009|
Conventional kinesin holoenzymes are composed of heavy and light chain homodimers.
PA1-642 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|