This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SRGN / Serglycin · Western blot design guide
Design a Western Blot for SRGN
Source-linked SRGN Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SRGN 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
SRGN Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~17.7 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SRGN Western Blot Protocol Options
The A06069 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A06069) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A06069) |
| Primary antibody | A06069 · 1:1000 (catalog A06069) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A06069) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A06069) |
Section 2
What Is the Expected SRGN Western Blot Band Size?
SRGN has a predicted 17.7 kDa precursor; signal-peptide cleavage and O-linked glycosaminoglycans could affect migration, but no empirical band position is supplied.
| Band near 17.7 kDa | Compatible with the predicted full-length precursor; confirm its identity |
| Band below 17.7 kDa | May reflect removal of the 27-residue signal peptide |
| Higher band or broad smear | May reflect variable O-linked glycosaminoglycans on serglycin |
| Several bands at different positions | May reflect differences in signal-peptide processing or glycosaminoglycan substitution |
| Little or no band in whole-cell lysate | Serglycin can be secreted into extracellular space |
💡Expected SRGN appearanceUniProt predicts a 17.7 kDa precursor, but signal-peptide cleavage and O-linked glycosaminoglycans may change migration; no empirical band size is supplied, so confirm candidate bands with identity controls.
| UniProt predicted precursor mass | Sets a 17.7 kDa sequence-based reference, not a validated blot position |
| Signal peptide at residues 1–27 | Its removal produces a smaller mature protein; apparent mass is unspecified |
| O-linked glycosaminoglycan sites at Ser94 and Ser96 | Substitution may increase or vary apparent mass |
| O-linked glycosaminoglycan sites at Ser100 and Ser102 | Substitution may increase or vary apparent mass |
| O-linked glycosaminoglycan sites at Ser104 and Ser106 | Substitution may increase or vary apparent mass |
| O-linked glycosaminoglycan sites at Ser108 and Ser110 | Substitution may increase or vary apparent mass |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Serglycin may be secreted or retained in granules | Check conditioned medium and a granule-containing fraction alongside lysate |
| Band higher than expected | O-linked glycosaminoglycan substitution may alter migration | Compare untreated and glycosaminoglycan-digested samples with an identity control |
| Band lower than expected | Signal-peptide removal can reduce protein size | Check whether an independent SRGN antibody detects the same band |
| Broad smear instead of sharp band | Variable glycosaminoglycan substitution is possible | Compare untreated and glycosaminoglycan-digested samples |
| Multiple bands | Processing or glycosaminoglycan substitution may differ among molecules | Compare fractions and verify bands with an independent SRGN antibody |
| Weak or no signal | Serglycin may be distributed between granules and extracellular medium | Examine both cellular and conditioned-medium samples |
Experiment evidence
Western blot images showing SRGN band
A06069 · SRGN
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SRGN Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SRGN in Western blot, you can use bone marrow tissue, which HPA rates highly positive.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Because SRGN is secreted and granule-associated, lysate signal may vary with sample preparation.
HPA tissue expression evidence for SRGN
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Skin | lymphocytes | High | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | High | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Lung | macrophages | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SRGN Western Blot Tips
Deeper troubleshooting and optimisation questions for SRGN, answered from its protein features.
How should SRGN 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 SRGN isoforms explain multiple bands?
Isoforms · The supplied features list one isoform and no alternative sequence. Do not assign additional bands to an annotated SRGN isoform on this evidence alone. Assess signal peptide processing and glycosaminoglycan modification when interpreting them.
Which SRGN modifications matter when interpreting a Western blot?
PTM · UniProt annotates O-linked glycosaminoglycan attachment at serines 94, 96, 100, 102, 104, 106, 108, and 110. These are UniProt sequence coordinates; paper or antibody numbering may differ. Consider these sites when assessing heterogeneous migration, but their annotation does not establish a visible shift.
Does this guide establish induction of SRGN?
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 SRGN?
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 A06069 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.
How should SRGN bands be quantified across samples?
Quantitation · Compare bands from the same sample fraction and preparation, since SRGN is annotated in intracellular compartments and extracellular space. If measuring both cellular and secreted SRGN, report those pools separately rather than treating one as the total.
Why might SRGN migrate differently from its predicted 17.7 kDa mass?
Interpretation · The 17.7 kDa prediction describes the unmodified sequence. SRGN has a signal peptide at residues 1–27 and eight annotated glycosaminoglycan attachment sites, either of which may affect the species measured. These features alone cannot establish an apparent band size or explain a particular mass difference; no observed band is supplied.
SRGN has a signal peptide at residues 1–27. A processed species may differ from the full 158-residue sequence used for the 17.7 kDa prediction. The supplied features do not establish its apparent migration, so avoid assigning a band solely from calculated mass.
UniProt places SRGN in cytoplasmic and cytolytic granules, the Golgi apparatus, and extracellular space. Choose the sample fraction for the pool you intend to measure, and keep that fraction consistent across comparisons.
UniProt lists one disulfide bond. Comparing matched reducing and nonreducing preparations may help assess whether disulfide-dependent structure contributes to differing migration. The annotation alone does not predict a particular band pattern.
SRGN has a signal peptide at residues 1–27. A processed species may differ from the full 158-residue sequence used for the 17.7 kDa prediction. The supplied features do not establish its apparent migration, so avoid assigning a band solely from calculated mass.
UniProt places SRGN in cytoplasmic and cytolytic granules, the Golgi apparatus, and extracellular space. Choose the sample fraction for the pool you intend to measure, and keep that fraction consistent across comparisons.
UniProt lists one disulfide bond. Comparing matched reducing and nonreducing preparations may help assess whether disulfide-dependent structure contributes to differing migration. The annotation alone does not predict a particular band pattern.
Boster reagents
SRGN 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-Serglycin SRGN Antibody
Cat # A06069
The catalog reports A06069, an anti-SRGN antibody with stated human and mouse reactivity. Its WB image shows extracts from various cell lines at 1:1000 dilution, with 25 µg protein per lane; the caption does not identify the cell lines.
Which to pick: A06069 is the only listed option. It has a WB image and stated human and mouse reactivity; check whether its reported cell-line conditions fit your samples.
Source: BosterBio SRGN gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P10124.
- Human Protein Atlas. SRGN tissue expression.
- PMC12748857 — target-verified WB comparison
- PMC7957973 — target-verified WB comparison
- PMC12080381 — target-verified WB comparison