Enter your Stock Antibody Concentration, Desired Final Concentration, and Total Volume Needed, then set your Starting Dilution and Dilution Factor to find the Antibody Volume to pull, plus Buffer Volume, Dilution Ratio, and Calculated Titer.
Antibody Volume Needed
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Buffer Volume Needed
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Dilution Ratio
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Calculated Titer
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An antibody titer is the highest dilution of an antibody solution that still produces a detectable signal. It helps determine the optimal concentration for experiments and ensures reproducible results while minimizing background noise.
Start with the manufacturer's recommended dilution range, typically 1:100 to 1:1000 for primary antibodies. If no recommendation exists, begin with 1:100 and perform a serial dilution to find the optimal concentration.
Primary antibodies bind directly to your target protein and are typically used at 1:100-1:5000 dilutions. Secondary antibodies bind to primary antibodies and are usually more concentrated, requiring higher dilutions of 1:1000-1:10000.
Include 6-8 dilution points spanning 2-3 log orders to capture the full dynamic range. This ensures you find both the optimal working dilution and the point where signal drops to background levels.
Use 2-fold or 3-fold dilutions for fine titration when working near the optimal range. Use 5-fold or 10-fold dilutions for broader screening or when you need to cover a wider concentration range quickly.
Avoid storing diluted antibodies when possible, as dilution reduces stability. If storage is necessary, add BSA or glycerol as stabilizers, store at 4°C, and use within 24-48 hours for best results.
Use the formula C1×V1 = C2×V2, where C1 is stock concentration, V1 is stock volume needed, C2 is final concentration, and V2 is final volume. This calculator automates this calculation for you.