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- Table of Contents
STC2 / Stanniocalcin-2 · Western blot design guide
Design a Western Blot for STC2
Real validated STC2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-STC2 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
STC2 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 | ~33.2 kDa | |
| Observed band | ~33 kDa | |
| Gel | 5–20% (catalog A05017) | |
| Positive control | Adrenal gland (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 + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated STC2 Western Blot Protocols
The A05017 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human Caco-2, rat PC-12 (catalog A05017) |
| Gel % | 5–20% (catalog A05017) |
| Load | 30 ug; reducing conditions (catalog A05017) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05017) |
| Membrane | nitrocellulose membrane (catalog A05017) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05017) |
| Primary antibody | A05017 · 0.5 μg/mL (catalog A05017) |
| Primary incubation | overnight at 4°C (catalog A05017) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05017) |
| Secondary incubation | 1.5 hour at RT (catalog A05017) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05017) |
| Detection | ECL (catalog A05017) |
Section 2
What Is the Expected STC2 Western Blot Band Size?
STC2 is predicted at 33.2 kDa and observed at ~33 kDa; these values agree approximately, while feature-dependent migration effects remain unproven.
| Band at ~33 kDa under reducing conditions | Matches the empirical STC2 band and is close to the 33.2 kDa predicted precursor mass. |
| Band near twice the monomer size under nonreducing conditions | Could represent the disulfide-linked STC2 homodimer. |
| Band below the precursor position | Could reflect removal of the 1–24 signal peptide; its mature band position is not supplied. |
| Little or no band in whole-cell lysate | STC2 is secreted and may be more readily detected in conditioned medium. |
💡Expected STC2 appearanceSTC2 has a predicted precursor mass of 33.2 kDa and an empirical band at ~33 kDa under reducing conditions; confirm band identity with an appropriate control.
| Predicted precursor mass | 33.2 kDa provides a reference for the empirical ~33 kDa band. |
| N-linked glycosylation at Asn73 | May affect apparent mass, but the site alone does not establish a visible shift. |
| Disulfide-linked homodimer | May appear near twice the monomer mass if disulfide bonds remain intact. |
| Signal peptide at residues 1–24 | Its removal makes mature STC2 smaller than the precursor; no mature band size is supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Secreted STC2 may be depleted from whole-cell lysate. | Probe conditioned medium and use a band-identity control. |
| Band higher than expected | Incomplete reduction may preserve disulfide-linked homodimers. | Compare reducing and nonreducing samples. |
| Band lower than expected | Signal-peptide removal could lower the mass relative to the precursor. | Check band identity with STC2 depletion or another specific control. |
| Broad smear instead of sharp band | Variation in N-linked glycosylation at Asn73 is possible; the smear cause is unproven. | Compare untreated and deglycosylated samples with an identity control. |
| Multiple bands | Monomer and incompletely reduced homodimer may coexist. | Repeat with complete reduction and confirm STC2 identity. |
| Weak or no signal | Secretion may leave little STC2 in the sampled cell fraction. | Concentrate conditioned medium and include a positive control. |
Experiment evidence
Western blot images showing STC2 band
A05017 · STC2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for STC2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for STC2 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: STC2 is secreted, so conditioned medium may give a stronger signal than whole-cell lysate.
HPA tissue expression evidence for STC2
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | 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 → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced STC2 Western Blot Tips
Deeper troubleshooting and optimisation questions for STC2, answered from its protein features.
How should STC2 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 STC2 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Assess other listed features, including glycosylation, phosphorylation, signal peptide processing, and disulfide-linked homodimerization, without assigning a band to any one feature solely from its presence.
Could glycosylation affect STC2 band interpretation?
PTM · UniProt lists one N-linked glycosylation site, Asn73, in canonical STC2 numbering. Consider glycosylation when assessing band variation, but the listed site alone does not establish whether a shift will be visible.
UniProt lists Ser250 and Ser251 as phosphoserines and Thr254 as a phosphothreonine, all modified by FAM20C. These are canonical sequence coordinates; antibody or paper numbering may differ. Their presence does not establish a visible mobility shift.
UniProt lists Ser250 and Ser251 as phosphoserines and Thr254 as a phosphothreonine, all modified by FAM20C. These are canonical sequence coordinates; antibody or paper numbering may differ. Their presence does not establish a visible mobility shift.
Does this guide establish induction of STC2?
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 STC2?
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 A05017 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.
Where should STC2 be sampled for quantitation?
Quantitation · STC2 is annotated as secreted and has a signal peptide at residues 1–24. If measuring secreted STC2, include the extracellular sample in the collection plan. Keep sample type and collection conditions consistent across comparisons.
Why is the STC2 band near its predicted mass?
Interpretation · The observed band is approximately 33 kDa, close to the 33.2 kDa predicted mass. STC2 has a signal peptide at residues 1–24 and an N-linked glycosylation site at Asn73. These features alone do not establish a visible shift or explain the precise apparent mass.
UniProt describes STC2 as a disulfide-linked homodimer. Compare reducing and nonreducing samples when evaluating a higher apparent mass band; a change would support involvement of disulfide-linked species, but band position alone does not identify the species.
STC2 has a signal peptide spanning residues 1–24. Processing could change the length of the detected protein, but the feature alone does not establish the position or identity of a smaller band. State whether a mass estimate uses the full 302-residue sequence or the sequence after signal peptide removal.
UniProt describes STC2 as a disulfide-linked homodimer. Compare reducing and nonreducing samples when evaluating a higher apparent mass band; a change would support involvement of disulfide-linked species, but band position alone does not identify the species.
STC2 has a signal peptide spanning residues 1–24. Processing could change the length of the detected protein, but the feature alone does not establish the position or identity of a smaller band. State whether a mass estimate uses the full 302-residue sequence or the sequence after signal peptide removal.
Boster reagents
STC2 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-Stanniocalcin 2/STC2 Antibody Picoband®
Cat # A05017
A05017 is a rabbit polyclonal anti-STC2 antibody with reported human, mouse, and rat reactivity. Its supplied Western blot image shows an approximately 33 kDa band in MCF-7, Caco-2, PC-12, and Raw264.7 whole-cell lysates; no independent validation is supplied.
Which to pick: A05017 is the only listed option. Its Western blot image includes human, rat, and mouse cell lysates under reducing conditions, making those reported sample contexts the practical starting point for choosing it.
Source: BosterBio STC2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O76061.
- Human Protein Atlas. STC2 tissue expression.
- PMC10375666 — target-verified WB comparison