Design a Western Blot for MATR3

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 two isoforms. In isoform 2, residues 1–304 of the canonical sequence are replaced by a 16-residue sequence, so the isoforms could migrate differently. Compare bands with antibodies recognizing a region shared by both isoforms, such as canonical residues 305–847.

Check its epitope against the isoform sequences. Isoform 2 replaces canonical residues 1–304, which contain several listed phosphosites, including Ser-4 and Tyr-158. A reagent targeting a site in that replaced region should not be assumed to recognize isoform 2. Site numbers here use UniProt canonical coordinates.

PTM · UniProt lists 44 modified residues, including phosphorylation at Ser-4, Thr-150 and Tyr-158, and acetylation at Ser-2 and Lys-571. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. The listed sites do not establish that a particular modification produces a visible band shift.

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 A02931-2 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 · 94.6 kDa is the predicted mass; the reported Western blot band is about 125 kDa. UniProt lists phosphorylation and acetylation sites, but their presence alone does not explain this difference. Use the observed band position as a reference and verify band identity independently.

MATR3 is annotated in the nucleus matrix. Keep sample preparation and fraction choice consistent across samples, then quantify the same validated band in each lane. If multiple bands are present, determine which represent MATR3 before combining their signals.

Isoform 2 is a candidate because it replaces canonical residues 1–304 with 16 residues. Band position alone cannot establish its identity. Check whether the antibody recognizes a sequence retained in isoform 2 and compare the candidate band with the canonical MATR3 band.

Not by itself. UniProt lists phosphorylation, acetylation and a Ubl-conjugation keyword, but the supplied features do not assign a modification to a band or specify a Ubl-conjugation site. Validate the band's identity before interpreting its migration as a modification.