i think this will help you,
<asp:textbox id="TextBox1" runat="server"></asp:textbox>
<asp:requiredfieldvalidator id="RequiredFieldValidator1" runat="server" errormessage="1st one" validationgroup="1st" controltovalidate="TextBox1">
</asp:requiredfieldvalidator>
<asp:button id="Button1" runat="server" text="Button" validationgroup="1st" causesvalidation="true" /><br />
<asp:textbox id="TextBox2" runat="server"></asp:textbox>
<asp:requiredfieldvalidator id="RequiredFieldValidator2" runat="server" errormessage="2nd one" validationgroup="2nd" controltovalidate="TextBox2">
</asp:requiredfieldvalidator>
<asp:button id="Button2" runat="server" text="Button" validationgroup="2nd" causesvalidation="true" />