WebResource.axd Error 500 & Webform_OnSubmit method issue while validating.

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ASP.NET

IIS

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I am trying to deploy a web application on Window Server 2008 R2,

I have some Required field validator's and some Regular Expression validators on the page.
and I am validating those through validation summary by javascript in Webform_onsubmit method.

but problem is that , neither that method is working properly and unable to show the validation summary with validators.

Further I have found "WebResource.axd Internal Server Error 500" in Network tab of Chrome Debugger.

after googling alot, yet I am unable to find any solution.

Can anybody guide me please???

NOTE: I am running application from VS 2010 and also tried after deploying on IIS 7 but still the same issue.