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- Table of Contents
GRN / Progranulin · Western blot design guide
Design a Western Blot for GRN
Real validated GRN Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GRN WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
GRN Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~63.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated GRN Western Blot Protocols
The A00893 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of A-431 cells, (catalog A00893) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A00893) |
| Primary antibody | A00893 · 1:1000 (catalog A00893) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A00893) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A00893) |
Section 2
What Is the Expected GRN Western Blot Band Size?
Full-length GRN is predicted at 63.5 kDa; glycosylation, cleavage, homodimerization and isoforms may affect bands, but their migration is not demonstrated here.
| Band near 63.5 kDa | consistent with the predicted full-length precursor; confirm identity with controls |
| Band above 63.5 kDa | could reflect N-linked glycosylation; migration is not established |
| Higher band near twice the monomer size | could reflect a homodimer if it survives electrophoresis |
| Band below 63.5 kDa | could reflect signal-peptide cleavage or proteolysis |
| Several bands at different positions | could reflect isoforms 1, 2 and 3 or proteolysis; their separation is not established |
| Little or no band in whole-cell lysate | may reflect secretion, although progranulin is also lysosomal |
💡Expected GRN appearanceThe predicted full-length mass is 63.5 kDa; no empirical band size is supplied, and glycosylation, signal-peptide cleavage or proteolysis could alter migration, so confirm band identity with appropriate controls.
| Predicted full-length mass | 63.5 kDa provides the precursor reference size |
| N-linked glycosylation at Asn118 | could increase apparent size; no shift is established |
| N-linked glycosylation at Asn236 | could increase apparent size; no shift is established |
| N-linked glycosylation at Asn265 | could increase apparent size; no shift is established |
| N-linked glycosylation at Asn368 | could increase apparent size; no shift is established |
| N-linked glycosylation at Asn530 | could increase apparent size; no shift is established |
| Secreted homodimer | could appear near twice the monomer size if the dimer survives electrophoresis |
| Isoforms 1, 2 and 3 | may differ in size, but their masses and separation are not supplied |
| Signal peptide at residues 1–17 | cleavage makes mature progranulin smaller than the full-length precursor |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | progranulin is secreted | check conditioned medium alongside lysate with a positive control |
| Band higher than expected | N-linked glycosylation or a homodimer that survives electrophoresis | compare deglycosylated and untreated samples, and check reducing denaturing conditions |
| Band lower than expected | signal-peptide cleavage or proteolysis | compare fresh samples with a positive control and check which region the antibody recognizes |
| Broad smear instead of sharp band | heterogeneous N-linked glycosylation is possible | compare deglycosylated and untreated samples |
| Multiple bands | isoforms or proteolytic products are possible | compare antibody epitopes and a positive control; assess sample degradation |
| Fragments below expected size | progranulin can undergo proteolysis | process samples promptly and compare with protease-inhibited samples |
Experiment evidence
Western blot images showing GRN band
A00893 · GRN
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GRN Western blot
🧪For positive controls for GRN in Western blot, you can use a validated GRN-positive sample; the supplied HPA data identify no positive tissue or cell line.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: GRN is secreted, so conditioned medium may be useful; the supplied HPA data cannot establish tissue controls.
HPA tissue expression evidence for GRN
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced GRN Western Blot Tips
Deeper troubleshooting and optimisation questions for GRN, answered from its protein features.
How should GRN band migration be interpreted?
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.
Could GRN isoforms produce different bands?
Isoforms · Three isoforms are listed. Relative to canonical isoform 1, isoform 2 lacks residues 377–531; isoform 3 replaces residues 1–71 and lacks residues 72–251. These UniProt coordinates describe canonical sequence positions. Check which isoform your sample and antibody can detect before assigning bands.
Which GRN glycosylation sites matter when interpreting bands?
PTM · UniProt lists N-linked sites at residues 118, 236, 265, 368, and 530, using canonical sequence coordinates. Consider glycosylation when comparing band patterns, but do not assign any band to a particular site from position alone.
Does this guide establish induction of GRN?
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.
How should transfer be checked for GRN?
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.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00893 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.
What should GRN quantitation account for?
Quantitation · Define the band and sample compartment consistently. GRN has three isoforms, five annotated N-linked sites, and secreted and lysosomal locations; these features can complicate comparisons across bands or sample types. Quantify only bands whose identity is supported in the assay.
Why might GRN migrate differently from its predicted 63.5 kDa?
Interpretation · The 63.5 kDa prediction does not account for processing of the 1–17 signal peptide or the five annotated N-linked glycosylation sites. These features can affect migration, but they do not establish a visible shift or a specific apparent mass. No observed band size was supplied.
GRN has alternative isoforms, and its interaction with SLPI protects it from proteolysis. Either isoform differences or cleavage could be relevant to a lower band. The supplied features do not identify a specific cleavage product, so confirm band identity before labeling it.
UniProt places GRN in both secreted and lysosomal compartments and notes delivery to lysosomes after endocytosis by SORT1. Specify the sample compartment when comparing signals; a change in one compartment alone does not establish a change in total GRN.
UniProt lists 12 disulfide bonds, so reduction may change how GRN migrates. Its reported secreted homodimer does not establish a disulfide-linked dimer. Compare reducing and nonreducing lanes if a higher band needs investigation, without assigning its identity from migration alone.
GRN has alternative isoforms, and its interaction with SLPI protects it from proteolysis. Either isoform differences or cleavage could be relevant to a lower band. The supplied features do not identify a specific cleavage product, so confirm band identity before labeling it.
UniProt places GRN in both secreted and lysosomal compartments and notes delivery to lysosomes after endocytosis by SORT1. Specify the sample compartment when comparing signals; a change in one compartment alone does not establish a change in total GRN.
UniProt lists 12 disulfide bonds, so reduction may change how GRN migrates. Its reported secreted homodimer does not establish a disulfide-linked dimer. Compare reducing and nonreducing lanes if a higher band needs investigation, without assigning its identity from migration alone.
Boster reagents
GRN Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Granulins GRN Antibody
Cat # A00893
A00893 is listed for human and mouse GRN and includes a Western blot image from A-431 cell extracts. The supplied evidence shows one sample context; it does not establish performance across other specimens.
Which to pick: A00893 is the only listed option. Its WB image uses A-431 extracts at 25 µg per lane and a 1:1000 primary dilution. Mouse reactivity is listed, but the supplied caption does not show a mouse sample.
Source: BosterBio GRN gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P28799.
- Human Protein Atlas. GRN tissue expression.
- PMC2581433 — target-verified WB comparison
- PMC11496013 — target-verified WB comparison
- PMC10312473 — target-verified WB comparison