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STING1 / Stimulator of interferon genes protein · Western blot design guide
Design a Western Blot for STING1
Source-linked STING1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-STING1 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
STING1 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 | ~42.2 kDa | |
| Observed band | ~36 kDa | |
| Gel | 10% (catalog A01871-3) | |
| Positive control | Bronchus (IHC candidate; verify WB) +4 more | |
| Negative control | Adrenal gland (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked STING1 Western Blot Protocol Options
The A01871-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, mouse spleen, mouse thymus, mouse MH-S (catalog A01871-3) |
| Gel % | 10% (catalog A01871-3) |
| Load | 30 ug; reducing conditions (catalog A01871-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01871-3) |
| Membrane | nitrocellulose membrane (catalog A01871-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01871-3) |
| Primary antibody | A01871-3 · 0.5 μg/mL (catalog A01871-3) |
| Primary incubation | overnight at 4°C (catalog A01871-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01871-3) |
| Secondary incubation | 1.5 hour at RT (catalog A01871-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01871-3) |
| Detection | ECL (catalog A01871-3) |
Section 2
What Is the Expected STING1 Western Blot Band Size?
STING1 is predicted at 42.2 kDa, but an empirical band appears near 36 kDa; the cause of this difference is not established.
| Band near 36 kDa | Empirical STING1 band reported in reducing whole-cell and tissue lysates |
| Band near 42 kDa | Near the 42.2 kDa predicted monomer mass; identity requires antibody controls |
| Band near twice the monomer mass | Possible homodimer if it survives sample preparation |
| Faint or absent band in lysate | ER membrane localization may limit recovery |
💡Expected STING1 appearanceSTING1 has a predicted mass of 42.2 kDa, while antibody QC reports a band near 36 kDa; the cause of this difference is unestablished, so confirm identity with antibody controls.
| Predicted monomer mass | 42.2 kDa predicted; the reported band is near 36 kDa |
| Homodimer formation | Could produce a band near twice the monomer mass if retained during preparation |
| Homodimer in the absence of cyclic nucleotide | May favor a higher-mass species if the dimer remains intact |
| Lys-63-linked ubiquitination at Lys-150 | Required for homodimerization, which could yield a higher-mass species if retained |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ER membrane protein may be poorly recovered | Check membrane extraction and a positive-control lysate |
| Band higher than expected | Homodimer may remain intact | Compare denaturing preparation and a validated STING1 control |
| Band lower than expected | The reported 36 kDa band runs below the 42.2 kDa prediction; the reason is unestablished | Compare the antibody QC control and verify band identity |
| Multiple bands | Monomer and retained homodimer are possible | Compare sample preparation conditions and antibody controls |
| Weak or no signal | Incomplete recovery of ER membrane protein | Check extraction efficiency and sample loading |
Experiment evidence
Western blot images showing STING1 band
A01871-3 · STING1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for STING1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for STING1 in Western blot, you can use bronchus tissue, which HPA rates as highly expressed.
Positive control: Bronchus (IHC candidate; verify WB)
Negative control: Adrenal gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: STING1 is an ER membrane protein, so use lysates that retain membrane proteins.
HPA tissue expression evidence for STING1
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 |
|---|---|---|---|---|
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Fallopian tube | ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Nasopharynx | basal cells | High | Protein (IHC) | HPA → |
| Skin | endothelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced STING1 Western Blot Tips
Deeper troubleshooting and optimisation questions for STING1, answered from its protein features.
How should STING1 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 additional STING1 bands represent isoforms?
Isoforms · This record lists one isoform and supplies no alternative sequence. Do not assign additional bands to documented isoforms on this evidence alone.
Which STING1 phosphorylation sites matter when probing a blot?
PTM · UniProt lists Thr229, Ser241, Thr354, Ser355 (by MAP3K7), Thr356, Ser358 (by TBK1), and Ser366 (by TBK1). These are UniProt coordinates; check the numbering used by the antibody or paper. Their presence does not establish a visible band shift.
Measure a site-specific phospho signal alongside total STING1 in matched samples, then compare their ratio. Keep the band assignment consistent across samples; listed phosphorylation sites alone do not establish protein abundance or a mobility shift.
Measure a site-specific phospho signal alongside total STING1 in matched samples, then compare their ratio. Keep the band assignment consistent across samples; listed phosphorylation sites alone do not establish protein abundance or a mobility shift.
Does this guide establish induction of STING1?
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.
What transfer method to use for STING1 Western blot?
Transfer · STING1 is an endoplasmic reticulum membrane protein with transmembrane regions. Check recovery on the membrane and retention in the gel when setting up transfer for its predicted 42.2 kDa mass. The supplied features do not identify a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01871-3 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 STING1 be quantified?
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.
Does the reported 36 kDa band match STING1's predicted mass?
Interpretation · STING1's predicted mass is 42.2 kDa, while the reported apparent band is about 36 kDa. The listed features do not establish the cause of that difference. Confirm the band's identity with a STING1-specific control before assigning it.
UniProt reports homodimerization and Lys-63-linked ubiquitination at Lys150 required for homodimerization. Those features suggest possibilities, but do not identify an extra band on a blot. Confirm STING1 specificity and test the proposed modification before labeling the band a dimer or ubiquitinated species.
UniProt reports homodimerization and Lys-63-linked ubiquitination at Lys150 required for homodimerization. Those features suggest possibilities, but do not identify an extra band on a blot. Confirm STING1 specificity and test the proposed modification before labeling the band a dimer or ubiquitinated species.
Boster reagents
STING1 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-TMEM173/STING Antibody Picoband®
Cat # A01871-3
The listed anti-STING1 antibody, A01871-3, reports human, mouse, and rat reactivity. Its Western blot image shows human Jurkat and mouse spleen, thymus, and MH-S lysates, with a band near 36 kDa versus an expected 42 kDa. No rat blot is shown.
Which to pick: A01871-3 is the only listed option. Its Western blot image provides examples for human Jurkat and three mouse samples. For rat samples, the listing reports reactivity but supplies no rat blot image.
Source: BosterBio STING1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q86WV6.
- Human Protein Atlas. STING1 tissue expression.
- PMC12175898 — target-verified WB comparison
- PMC10842782 — target-verified WB comparison
- PMC11410371 — target-verified WB comparison