I have bottleneck in 2nd antibody-QuantumDots for my Hela cells.
It is okey for me to use 2nd-antibody-rhodamine as positive control to detect my nulear antigens. But I fail in 2nd antibody-QuantumDots under the same conndition.
Is there one who has the experience in using 2nd-antibody-QuantumDots?
Here is my condition:
2.Primary antibody: purchased from Abcam
3.2nd antibody with rhodamine conjugated(goat anti-mouse): purchased from JacksonIR
4.2nd antibody with QuantumDots conjugated:Qdot 525 Goat F(ab')2 anti-Mouse IgG, purchased from Quantum Dots
5.Fixed HeLa with Histochoice followed by tritonX100
Most of the Quantumdots are in cytoplasm, in stead of nuclear.
If there is anyone with Quantum DOts experience, pls tell me how to solve my bottleneck.
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Problem with QuantumDots lebleing
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