Immunofluorescence analysis of CRMP5 was done on 70% confluent log phase SH-SY5Y 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 CRMP5 (CR-3) Rat Monoclonal Antibody (MA3700) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A11006) 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, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rat / IgG2a|
|Immunogen||CRMP5 protein purified from human brain.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:50 - 1:400|
|Western Blot (WB)||1:50 - 1:1,000|
* 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.
|Immunohistochemistry (IHC)||See 1 publications below|
MA3-700 detects human, bovine, rat and mouse N-terminal region of CRMP5.
MA3-700 has been successfully used in Western blot and immunohistochemical procedures. By Western blot, this antibody detects an ~ 62 kDa protein corresponding to human CRMP5 from brain extract.
The MA3-700 antigen is a purified protein from human brain. This antibody is specific for the N-terminal region, amino acids 1-64. Not reactive with other CRMP isoforms.
The collapsin response mediator protein (CRMP) family of proteins (also referred to as ULIP6) are a group of neurologic proteins that have been found to play key roles in growth cone guidance during neural development. CRMP5 has relatively low sequence homology with the other four members of the CRMP family. CRMP5 is expressed in the developing nervous system in an expression pattern that resembles CRMP2 and has a peak expression during the first postnatal week. Elevated levels of autoantibody to CRMP5 have been shown to be linked to paraneoplastic autoimmune optic neuritis and other paraneoplastic neoplasms.
Auto-immunity against CRMP5 expression (predominantly the N-terminal epitopes) is also being investigated as an oncological marker in the respiratory system and in the thymus.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Paraneoplastic autoimmune optic neuritis with retinitis defined by CRMP-5-IgG.
MA3-700 was used in immunohistochemistry to study the prognostic utility of CRMP-5-IgG in paraneoplastic autoimmune optic neuritis
|Cross SA,Salomao DR,Parisi JE,Kryzer TJ,Bradley EA,Mines JA,Lam BL,Lennon VA||Annals of neurology (54:38)||2003|