Design a Western Blot for LAMP2

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 · LAMP-2A, LAMP-2B and LAMP-2C are listed. LAMP-2B replaces residues 367–410 and LAMP-2C replaces residues 366–410 in UniProt coordinates. Check whether the antibody epitope lies in these differing regions before comparing isoform signals.

PTM · UniProt lists N-linked sites, including polylactosaminoglycan annotations at residues 32, 38 and 307, plus O-linked sites at residues 195–213. O-linked sites 207, 209 and 213 are annotated as partial. These are UniProt sequence coordinates. The annotations do not establish which sites are occupied in your sample or whether they cause a visible 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 A01573 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 · Use a consistent band selection and normalization approach across samples. LAMP2 has three isoforms and 26 annotated glycosylation sites, so confirm that the antibody recognizes the forms you intend to compare. The supplied record gives no empirical band position for defining an integration region.

Interpretation · The 45 kDa value is a predicted mass. UniProt lists 26 glycosylation sites and a signal peptide at residues 1–28, both relevant when interpreting migration. These features alone cannot establish an apparent band size or explain a specific difference; no observed band was supplied.

UniProt annotates a signal peptide at residues 1–28. If an antibody targets this region, check whether its epitope is present in the processed protein. Use UniProt sequence coordinates when comparing antibody and feature annotations.

UniProt reports LAMP2 as a monomer, homodimer, homotrimer and member of larger homooligomers. These associations make oligomers a possibility to investigate, but the feature record does not show that any particular Western-blot band is an oligomer. Compare bands under your sample preparation conditions before assigning them.

Check antibody epitope coverage of the isoform-specific regions at residues 366–410, consider the annotated glycosylation sites, and compare with the 1–28 signal peptide annotation. UniProt also reports oligomers. None of these features alone identifies an observed band, and no empirical band size was supplied.