Design a Western Blot for NUP153

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 · UniProt lists three isoforms. Relative to the canonical sequence, isoform 2 lacks residues 574–615, while isoform 3 replaces E465 with ERQGLTVLPKLISSSCAQAIIPSWPLKVLRLQ. Check whether the antibody epitope is retained in each isoform before interpreting multiple bands.

In canonical UniProt numbering, phosphothreonine 588 and phosphoserines 607 and 614 lie within residues 574–615, which are missing from isoform 2. Check isoform identity before interpreting signals from antibodies directed at these sites. Coordinates for residues after the deletion differ between isoforms.

PTM · UniProt lists N-acetylalanine at position 2 and N6-acetyllysine at positions 384, 718 and 954, using canonical coordinates. If an antibody recognizes a region containing one of these residues, check whether its validation covers the modified form. The site listings alone do not show that acetylation changes the Western blot band.

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 · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.

Blocking · Standard workflow guidance: follow the M02183 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 · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.

Interpretation · 153.9 kDa is the predicted mass; no observed band position is supplied. UniProt lists phosphorylation, acetylation and O-linked GlcNAc, but their presence alone does not establish a visible shift or explain any difference from predicted mass. Confirm band identity before assigning a cause.

UniProt lists O-linked GlcNAc at serines 534, 544, 908, 909 and 1113, and threonine 1156. These are canonical UniProt coordinates; confirm the numbering convention used for any site-specific antibody. These annotations alone do not predict a visible band shift.

NUP153 is listed at the nucleus, nuclear membrane and nuclear pore complex. Compare samples prepared with the same fractionation method, since different fractions may contain different amounts of NUP153. Use the same band identification criteria across samples, especially if multiple bands appear.

First compare them with the predicted 153.9 kDa mass and check antibody epitope coverage across the three isoforms. Isoform 2 lacks canonical residues 574–615; isoform 3 changes the sequence at 465. Listed modifications may affect detection, but the supplied features do not establish the identity or migration of an unexpected band.