Design a Western Blot for CXCL11

Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.

Isoforms · Only one isoform is supplied, with no alternative sequence. Do not assign additional bands to annotated isoforms on this evidence. Check their identity experimentally before interpreting them as CXCL11.

PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.

Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.

Transfer · CXCL11 is small: its supplied predicted mass is 10.4 kDa, and it has a 1–21 signal peptide. Use transfer conditions suited to retaining small proteins, such as a 0.2 µm membrane, and verify transfer and retention with appropriate controls.

Blocking · Standard workflow guidance: follow the M01833 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.

Quantitation · CXCL11 is annotated as secreted. For conditioned-medium samples, keep collection time and medium volume consistent, and normalize the signal to a relevant measure of the producing cells. State whether the reported signal comes from medium or cell lysate.

Interpretation · The observed band is about 15 kDa, while the supplied predicted mass is 10.4 kDa. CXCL11 has a signal peptide at residues 1–21, but its cleavage would reduce the mature protein’s calculated mass. The listed features alone do not explain the higher apparent mass; confirm band identity before assigning a cause.

CXCL11 is secreted and has a signal peptide at residues 1–21. A processed form would lack those residues, so compare any mass estimate with the appropriate sequence. Signal peptide cleavage alone does not account for an apparent band near 15 kDa.

UniProt lists citrullination by PAD2 at position 27, numbered in the full 94-residue sequence. If numbering starts after removal of the 1–21 signal peptide, that corresponds to mature residue 6. This feature establishes a modification site, but does not establish a visible band shift.

UniProt lists two disulfide bonds. Compare reducing and nonreducing samples if band behavior changes between preparations. The feature alone does not establish that either condition produces a particular band or accounts for the observed 15 kDa signal.

CXCL11 interacts with TNFAIP6 through its Link domain, but that annotation does not show that a complex persists during Western blotting. Verify unexpected bands with suitable specificity controls and compare reducing conditions before attributing them to CXCL11 or its interaction partner.