Immunofluorescence analysis of CRMP2 was performed using 70% confluent log phase Neuro-2a cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CRMP2 Rabbit Polyclonal Antibody (PA3120) at 2ug/ml in 0.1% 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 (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 represents the merged image showing cytoplasmic and membranous localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Mouse, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues Y(499) D G P V F D L T T T P K(511) of TUC-4a (residues 612-624 of TUC-4b).|
|Storage buffer||whole serum|
|Contains||0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||1:2000|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1-3 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.
|Miscellaneous PubMed (MISC)||See 2 publications below|
PA3-120 detects TOAD-64 in rat samples.
PA3-120 has been successfully used in Western blot, immunoprecipitation and immunohistochemistry (frozen) procedures. By Western blot, this antibody detects an 64 kDa protein representing TOAD-64 in rat samples.
The PA3-120 immunogen is a synthetic peptide corresponding to residues Y(499) D G P V F D L T T T P K(511) of TUC-4a (residues 612-624 of TUC-4b).
TOAD-64 is a 64 kDa intracellular neural protein, which belongs to a family of genes that inhibit nerve growth. It also appears to be involved in a role of axon outgrowth. Preliminary results indicate inhibition of TOAD-64 expression poteintiates neurite outgrowth, and may lead to the design of therapies that promote axonal regeneration. Antibodies to TOAD-64 render neurons insensitive to extracellular compounds that inhibit neuron growth. Very few reagents are able to identify newly born neurons as well as antibodies to TOAD-64.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
TOAD-64, a gene expressed early in neuronal differentiation in the rat, is related to unc-33, a C. elegans gene involved in axon outgrowth.
PA3-120 was used in immunohistochemistry and western blot to investigate the role of TOAD-64 in axonal outgrowth in vivo
|Minturn JE,Fryer HJ,Geschwind DH,Hockfield S||The Journal of neuroscience : the official journal of the Society for Neuroscience (15:6757)||1995|
|Not Applicable||Not Cited||
Early postmitotic neurons transiently express TOAD-64, a neural specific protein.
PA3-120 was used in immunohistochemistry, immunoprecipitation and western blot to investigate the expression and cellular distribution of TOAD-64 in postmitotic neurons during development
|Minturn JE,Geschwind DH,Fryer HJ,Hockfield S||The Journal of comparative neurology (355:369)||1995|