diff --git a/src/OpenTelemetry.Instrumentation.AspNetCore/AspNetCoreTraceInstrumentationOptions.cs b/src/OpenTelemetry.Instrumentation.AspNetCore/AspNetCoreTraceInstrumentationOptions.cs
index 6caadf3db90..f0fc396c969 100644
--- a/src/OpenTelemetry.Instrumentation.AspNetCore/AspNetCoreTraceInstrumentationOptions.cs
+++ b/src/OpenTelemetry.Instrumentation.AspNetCore/AspNetCoreTraceInstrumentationOptions.cs
@@ -17,9 +17,7 @@
 using System.Diagnostics;
 using Microsoft.AspNetCore.Http;
 using Microsoft.Extensions.Configuration;
-#if NET6_0_OR_GREATER
 using OpenTelemetry.Internal;
-#endif
 
 namespace OpenTelemetry.Instrumentation.AspNetCore;
 
@@ -40,12 +38,10 @@ internal AspNetCoreTraceInstrumentationOptions(IConfiguration configuration)
     {
         Debug.Assert(configuration != null, "configuration was null");
 
-#if NET6_0_OR_GREATER
-        if (configuration.TryGetBoolValue("OTEL_DOTNET_EXPERIMENTAL_ENABLE_GRPC_INSTRUMENTATION", out var enableGrpcInstrumentation))
+        if (configuration.TryGetBoolValue("OTEL_DOTNET_EXPERIMENTAL_ASPNETCORE_ENABLE_GRPC_INSTRUMENTATION", out var enableGrpcInstrumentation))
         {
             this.EnableGrpcAspNetCoreSupport = enableGrpcInstrumentation;
         }
-#endif
     }
 
     /// <summary>
@@ -101,7 +97,6 @@ internal AspNetCoreTraceInstrumentationOptions(IConfiguration configuration)
     /// </remarks>
     public bool RecordException { get; set; }
 
-#if NET6_0_OR_GREATER
     /// <summary>
     /// Gets or sets a value indicating whether RPC attributes are added to an Activity when using Grpc.AspNetCore. Default is true.
     /// </summary>
@@ -109,5 +104,4 @@ internal AspNetCoreTraceInstrumentationOptions(IConfiguration configuration)
     /// https://github.com/open-telemetry/semantic-conventions/blob/main/docs/rpc/rpc-spans.md.
     /// </remarks>
     internal bool EnableGrpcAspNetCoreSupport { get; set; }
-#endif
 }
diff --git a/src/OpenTelemetry.Instrumentation.AspNetCore/CHANGELOG.md b/src/OpenTelemetry.Instrumentation.AspNetCore/CHANGELOG.md
index 71d9a7bd189..6e71b160e4c 100644
--- a/src/OpenTelemetry.Instrumentation.AspNetCore/CHANGELOG.md
+++ b/src/OpenTelemetry.Instrumentation.AspNetCore/CHANGELOG.md
@@ -2,16 +2,18 @@
 
 ## Unreleased
 
-* Added support for gRPC instrumentation under an experimental feature flag
-`OTEL_DOTNET_EXPERIMENTAL_ENABLE_GRPC_INSTRUMENTATION`. Note that the support
-  was removed in `1.6.0-rc.1` version of the package and versions released
-  before `1.6.0-rc.1` had gRPC instrumentation enabled by default. From now
-  onwards, gRPC instrumentation will only be enabled when
-  `OTEL_DOTNET_EXPERIMENTAL_ENABLE_GRPC_INSTRUMENTATION` is set to `True`. The
-  change is introduced in order to support stable release of `http`
-  instrumentation. Semantic conventions for RPC is still
+* Re-introduced support for gRPC instrumentation as an opt-in experimental
+  feature. From now onwards, gRPC can be enabled by setting
+  `OTEL_DOTNET_EXPERIMENTAL_ASPNETCORE_ENABLE_GRPC_INSTRUMENTATION` to `True`.
+  `OTEL_DOTNET_EXPERIMENTAL_ASPNETCORE_ENABLE_GRPC_INSTRUMENTATION` can be set
+  as an environment variable or via IConfiguration. The change is introduced in
+  order to support stable release of `http` instrumentation. Semantic
+  conventions for RPC is still
   [experimental](https://github.com/open-telemetry/semantic-conventions/tree/main/docs/rpc)
-  and hence the package will only support it as an experimental feature.
+  and hence the package will only support it as an opt-in experimental feature.
+  Note that the support was removed in `1.6.0-rc.1` version of the package and
+  versions released before `1.6.0-rc.1` had gRPC instrumentation enabled by
+  default.
   ([#5130](https://github.com/open-telemetry/opentelemetry-dotnet/pull/5130))
 
 ## 1.6.0-rc.1
diff --git a/src/OpenTelemetry.Instrumentation.AspNetCore/TracerProviderBuilderExtensions.cs b/src/OpenTelemetry.Instrumentation.AspNetCore/TracerProviderBuilderExtensions.cs
index d61d5b6d4dd..c0cca518f40 100644
--- a/src/OpenTelemetry.Instrumentation.AspNetCore/TracerProviderBuilderExtensions.cs
+++ b/src/OpenTelemetry.Instrumentation.AspNetCore/TracerProviderBuilderExtensions.cs
@@ -76,9 +76,7 @@ public static TracerProviderBuilder AddAspNetCoreInstrumentation(
                 services.Configure(name, configureAspNetCoreTraceInstrumentationOptions);
             }
 
-#if !NETSTANDARD2_0
             services.RegisterOptionsFactory(configuration => new AspNetCoreTraceInstrumentationOptions(configuration));
-#endif
         });
 
         if (builder is IDeferredTracerProviderBuilder deferredTracerProviderBuilder)
diff --git a/test/OpenTelemetry.Instrumentation.Grpc.Tests/GrpcTests.server.cs b/test/OpenTelemetry.Instrumentation.Grpc.Tests/GrpcTests.server.cs
index 172c5e73300..b94e66f3159 100644
--- a/test/OpenTelemetry.Instrumentation.Grpc.Tests/GrpcTests.server.cs
+++ b/test/OpenTelemetry.Instrumentation.Grpc.Tests/GrpcTests.server.cs
@@ -58,7 +58,7 @@ public void GrpcAspNetCoreInstrumentationAddsCorrectAttributes(string enableGrpc
             .AddInMemoryCollection(new Dictionary<string, string>
             {
                 [SemanticConventionOptInKeyName] = "http",
-                ["OTEL_DOTNET_EXPERIMENTAL_ENABLE_GRPC_INSTRUMENTATION"] = enableGrpcAspNetCoreSupport,
+                ["OTEL_DOTNET_EXPERIMENTAL_ASPNETCORE_ENABLE_GRPC_INSTRUMENTATION"] = enableGrpcAspNetCoreSupport,
             })
             .Build();
 
@@ -131,7 +131,7 @@ public void GrpcAspNetCoreInstrumentationAddsCorrectAttributesWhenItCreatesNewAc
            .AddInMemoryCollection(new Dictionary<string, string>
            {
                [SemanticConventionOptInKeyName] = "http",
-               ["OTEL_DOTNET_EXPERIMENTAL_ENABLE_GRPC_INSTRUMENTATION"] = enableGrpcAspNetCoreSupport,
+               ["OTEL_DOTNET_EXPERIMENTAL_ASPNETCORE_ENABLE_GRPC_INSTRUMENTATION"] = enableGrpcAspNetCoreSupport,
            })
            .Build();