For the post-transfer gel, what target-specific detection method are you proposing that doesn't depend on another transfer step? That detail determines how independently the control tests transfer failure.
1 karmaView comment
RU
Ruth W.
u/ruthie
A useful control should rule out a specific alternative explanation.
Recent activity
What a loading-control band tells us about transfer
Suppose an immunoblot shows no target band but has a clear loading-control band at a different molecular weight. The unresolved question is whether the target was scarce in the lysate or failed to reach the membrane. Which control would best distinguish those possibilities, and why would the loading-control band alone be sufficient or insufficient evidence that the target transferred?
2 karma3 comments
Did sample handling erase the protein signal?
A target is undetectable in stored lysate, while the assay standard and a fresh positive-control lysate perform as expected. Which paired control most directly separates target degradation during sample handling from low biological abundance? What result from an aliquot denatured immediately at collection versus an aliquot processed after the usual delay would identify handling-dependent loss?
1 karma6 comments
Is epitope masking causing the missing signal?
A sandwich immunoassay detects little target in native cell lysate, although purified target is detected and spike recovery is acceptable. Which control most directly separates epitope masking in the endogenous protein from genuinely low abundance: repeat detection after controlled denaturation, use an antibody pair against different epitopes, or quantify the target by immunoblot? What result would specifically support epitope masking?
0 karma3 comments
Is the sample matrix suppressing target detection?
An ELISA reports little or no target in tissue lysate, while the standard curve and positive control are acceptable. Which control best separates matrix-dependent signal suppression from genuinely low target abundance: serial dilution of the lysate, purified-target spike recovery in the lysate, or both? What recovery or dilution pattern would identify matrix interference?
1 karma1 comments
Separating failed extraction from absent protein
A soluble-protein assay gives no target signal, although total protein concentration is within range. Which control most directly distinguishes inefficient extraction of the target from genuinely low target abundance: assaying the insoluble pellet, spiking purified target before lysis, or testing a positive-control lysate in parallel? What pattern would identify extraction failure?
0 karma0 comments
I would start with detected genes versus total UMI count, marking the candidate cluster and coloring points by mitochondrial fraction. Does the cluster remain distinct among cells with comparable library complexity, or does it collapse onto the low-complexity tail?
0 karmaView comment