diff --git a/python/packages/azure-ai/agent_framework_azure_ai/_client.py b/python/packages/azure-ai/agent_framework_azure_ai/_client.py
index cc00a34aa0..67bd0bae6a 100644
--- a/python/packages/azure-ai/agent_framework_azure_ai/_client.py
+++ b/python/packages/azure-ai/agent_framework_azure_ai/_client.py
@@ -155,7 +155,11 @@ def __init__(
         self.credential = async_credential
         self.model_id = azure_ai_settings.model_deployment_name
         self.conversation_id = conversation_id
-        self._should_close_client = should_close_client  # Track whether we should close client connection
+
+        # Track whether the application endpoint is used
+        self._is_application_endpoint = "/applications/" in project_client._config.endpoint  # type: ignore
+        # Track whether we should close client connection
+        self._should_close_client = should_close_client
 
     async def setup_azure_ai_observability(self, enable_sensitive_data: bool | None = None) -> None:
         """Use this method to setup tracing in your Azure AI Project.
@@ -316,9 +320,11 @@ async def prepare_options(
         """Take ChatOptions and create the specific options for Azure AI."""
         prepared_messages, instructions = self._prepare_input(messages)
         run_options = await super().prepare_options(prepared_messages, chat_options, **kwargs)
-        agent_reference = await self._get_agent_reference_or_create(run_options, instructions)
 
-        run_options["extra_body"] = {"agent": agent_reference}
+        if not self._is_application_endpoint:
+            # Application-scoped response APIs do not support "agent" property.
+            agent_reference = await self._get_agent_reference_or_create(run_options, instructions)
+            run_options["extra_body"] = {"agent": agent_reference}
 
         conversation_id = chat_options.conversation_id or self.conversation_id
 
diff --git a/python/packages/azure-ai/tests/test_azure_ai_client.py b/python/packages/azure-ai/tests/test_azure_ai_client.py
index 5d48e7b2be..8a31abcf35 100644
--- a/python/packages/azure-ai/tests/test_azure_ai_client.py
+++ b/python/packages/azure-ai/tests/test_azure_ai_client.py
@@ -87,6 +87,7 @@ def create_test_azure_ai_client(
     client.use_latest_version = use_latest_version
     client.model_id = azure_ai_settings.model_deployment_name
     client.conversation_id = conversation_id
+    client._is_application_endpoint = False  # type: ignore
     client._should_close_client = should_close_client  # type: ignore
     client.additional_properties = {}
     client.middleware = None
@@ -305,6 +306,84 @@ async def test_azure_ai_client_prepare_options_basic(mock_project_client: MagicM
         assert run_options["extra_body"]["agent"]["name"] == "test-agent"
 
 
+@pytest.mark.parametrize(
+    "endpoint,expects_agent",
+    [
+        ("https://example.com/api/projects/my-project/applications/my-application/protocols", False),
+        ("https://example.com/api/projects/my-project", True),
+    ],
+)
+async def test_azure_ai_client_prepare_options_with_application_endpoint(
+    mock_azure_credential: MagicMock, endpoint: str, expects_agent: bool
+) -> None:
+    client = AzureAIClient(
+        project_endpoint=endpoint,
+        model_deployment_name="test-model",
+        async_credential=mock_azure_credential,
+        agent_name="test-agent",
+        agent_version="1",
+    )
+
+    messages = [ChatMessage(role=Role.USER, contents=[TextContent(text="Hello")])]
+    chat_options = ChatOptions()
+
+    with (
+        patch.object(client.__class__.__bases__[0], "prepare_options", return_value={"model": "test-model"}),
+        patch.object(
+            client,
+            "_get_agent_reference_or_create",
+            return_value={"name": "test-agent", "version": "1", "type": "agent_reference"},
+        ),
+    ):
+        run_options = await client.prepare_options(messages, chat_options)
+
+    if expects_agent:
+        assert "extra_body" in run_options
+        assert run_options["extra_body"]["agent"]["name"] == "test-agent"
+    else:
+        assert "extra_body" not in run_options
+
+
+@pytest.mark.parametrize(
+    "endpoint,expects_agent",
+    [
+        ("https://example.com/api/projects/my-project/applications/my-application/protocols", False),
+        ("https://example.com/api/projects/my-project", True),
+    ],
+)
+async def test_azure_ai_client_prepare_options_with_application_project_client(
+    mock_project_client: MagicMock, endpoint: str, expects_agent: bool
+) -> None:
+    mock_project_client._config = MagicMock()
+    mock_project_client._config.endpoint = endpoint
+
+    client = AzureAIClient(
+        project_client=mock_project_client,
+        model_deployment_name="test-model",
+        agent_name="test-agent",
+        agent_version="1",
+    )
+
+    messages = [ChatMessage(role=Role.USER, contents=[TextContent(text="Hello")])]
+    chat_options = ChatOptions()
+
+    with (
+        patch.object(client.__class__.__bases__[0], "prepare_options", return_value={"model": "test-model"}),
+        patch.object(
+            client,
+            "_get_agent_reference_or_create",
+            return_value={"name": "test-agent", "version": "1", "type": "agent_reference"},
+        ),
+    ):
+        run_options = await client.prepare_options(messages, chat_options)
+
+    if expects_agent:
+        assert "extra_body" in run_options
+        assert run_options["extra_body"]["agent"]["name"] == "test-agent"
+    else:
+        assert "extra_body" not in run_options
+
+
 async def test_azure_ai_client_initialize_client(mock_project_client: MagicMock) -> None:
     """Test initialize_client method."""
     client = create_test_azure_ai_client(mock_project_client)
diff --git a/python/samples/getting_started/agents/azure_ai/README.md b/python/samples/getting_started/agents/azure_ai/README.md
index 0513dab1df..17a944524a 100644
--- a/python/samples/getting_started/agents/azure_ai/README.md
+++ b/python/samples/getting_started/agents/azure_ai/README.md
@@ -16,6 +16,7 @@ This folder contains examples demonstrating different ways to create and use age
 | [`azure_ai_with_code_interpreter.py`](azure_ai_with_code_interpreter.py) | Shows how to use the `HostedCodeInterpreterTool` with Azure AI agents to write and execute Python code for mathematical problem solving and data analysis. |
 | [`azure_ai_with_existing_agent.py`](azure_ai_with_existing_agent.py) | Shows how to work with a pre-existing agent by providing the agent name and version to the Azure AI client. Demonstrates agent reuse patterns for production scenarios. |
 | [`azure_ai_with_existing_conversation.py`](azure_ai_with_existing_conversation.py) | Demonstrates how to use an existing conversation created on the service side with Azure AI agents. Shows two approaches: specifying conversation ID at the client level and using AgentThread with an existing conversation ID. |
+| [`azure_ai_with_application_endpoint.py`](azure_ai_with_application_endpoint.py) | Demonstrates calling the Azure AI application-scoped endpoint. |
 | [`azure_ai_with_explicit_settings.py`](azure_ai_with_explicit_settings.py) | Shows how to create an agent with explicitly configured `AzureAIClient` settings, including project endpoint, model deployment, and credentials rather than relying on environment variable defaults. |
 | [`azure_ai_with_file_search.py`](azure_ai_with_file_search.py) | Shows how to use the `HostedFileSearchTool` with Azure AI agents to upload files, create vector stores, and enable agents to search through uploaded documents to answer user questions. |
 | [`azure_ai_with_hosted_mcp.py`](azure_ai_with_hosted_mcp.py) | Shows how to integrate hosted Model Context Protocol (MCP) tools with Azure AI Agent. |
diff --git a/python/samples/getting_started/agents/azure_ai/azure_ai_with_application_endpoint.py b/python/samples/getting_started/agents/azure_ai/azure_ai_with_application_endpoint.py
new file mode 100644
index 0000000000..89bb77af11
--- /dev/null
+++ b/python/samples/getting_started/agents/azure_ai/azure_ai_with_application_endpoint.py
@@ -0,0 +1,39 @@
+# Copyright (c) Microsoft. All rights reserved.
+
+import asyncio
+import os
+
+from agent_framework import ChatAgent
+from agent_framework.azure import AzureAIClient
+from azure.ai.projects.aio import AIProjectClient
+from azure.identity.aio import AzureCliCredential
+
+"""
+Azure AI Agent with Application Endpoint Example
+
+This sample demonstrates working with pre-existing Azure AI Agents by providing
+application endpoint instead of project endpoint.
+"""
+
+
+async def main() -> None:
+    # Create the client
+    async with (
+        AzureCliCredential() as credential,
+        # Endpoint here should be application endpoint with format:
+        # /api/projects/<project-name>/applications/<application-name>/protocols
+        AIProjectClient(endpoint=os.environ["AZURE_AI_PROJECT_ENDPOINT"], credential=credential) as project_client,
+        ChatAgent(
+            chat_client=AzureAIClient(
+                project_client=project_client,
+            ),
+        ) as agent,
+    ):
+        query = "How are you?"
+        print(f"User: {query}")
+        result = await agent.run(query)
+        print(f"Agent: {result}\n")
+
+
+if __name__ == "__main__":
+    asyncio.run(main())