This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SPRED1 / Sprouty-related, EVH1 domain-containing protein 1 · Western blot design guide
Design a Western Blot for SPRED1
Source-linked SPRED1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SPRED1 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
SPRED1 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 | ~50.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| 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 | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SPRED1 Western Blot Protocol Options
The A03613 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse brain tissue lysate (catalog A03613) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03613 · (A) 1 and (B) 2 μg/mL (catalog A03613) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected SPRED1 Western Blot Band Size?
SPRED1 has a predicted 50.5 kDa monomer; homodimerization and documented modifications are possible band factors, but their electrophoretic effects are not demonstrated here.
| Band near 50.5 kDa | consistent with the predicted SPRED1 monomer; confirm identity with antibody controls |
| Band near 101 kDa | could reflect a homodimer that remains associated during sample preparation |
| Bands near 50.5 and 101 kDa | could reflect monomer and retained homodimer |
| Weak band in whole-cell lysate | may reflect SPRED1 distribution between peripheral membranes and nucleus |
💡Expected SPRED1 appearanceUniProt predicts a 50.5 kDa SPRED1 monomer, but no empirical band size is supplied; use antibody specificity and band-identity controls to assess any observed band.
| UniProt predicted monomer mass | sets the full-length reference at 50.5 kDa |
| Homodimer formation | could yield a band near twice the monomer mass if the complex remains associated |
| N6-methyllysine at residue 224 | is documented, but no apparent-size effect is established |
| Phosphoserine at residues 238 and 308 | is documented, but no resolvable migration difference is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SPRED1 occupies peripheral membrane and nuclear compartments | Check membrane and nuclear fractions alongside whole-cell lysate |
| Band higher than expected | A SPRED1 homodimer may remain associated | Compare denaturing conditions and confirm band identity |
| Band lower than expected | The supplied features do not establish a smaller mature product | Check antibody specificity and sample integrity |
| Multiple bands | A retained homodimer could accompany the monomer; other band identities are unestablished | Compare denaturing conditions and use a SPRED1-specific identity control |
| Weak or no signal | SPRED1 may be underrepresented in the sampled cell fraction | Check membrane and nuclear fractions and verify antibody performance |
Experiment evidence
Western blot images showing SPRED1 band
A03613 · SPRED1
A03613-1 · SPRED1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SPRED1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SPRED1 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: SPRED1 is membrane-associated, so membrane enrichment may improve detection in tissue lysates.
HPA tissue expression evidence for SPRED1
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | 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 → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SPRED1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SPRED1, answered from its protein features.
How should SPRED1 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 an alternative SPRED1 isoform explain another band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning another band to a listed isoform. Verify an additional band before interpreting it as SPRED1.
Which SPRED1 modifications should guide band interpretation?
PTM · UniProt lists N-acetylserine at position 2, N6-methyllysine at 224, and phosphoserines at 238 and 308. These are UniProt coordinates; paper or antibody numbering may differ. None of these annotations alone establishes a resolvable Western-blot shift.
Phosphoserines are listed at UniProt positions 238 and 308, but no induction condition or treatment response is supplied. If a band changes after treatment, compare matched samples and verify phosphorylation before attributing the change to either site.
Phosphoserines are listed at UniProt positions 238 and 308, but no induction condition or treatment response is supplied. If a band changes after treatment, compare matched samples and verify phosphorylation before attributing the change to either site.
Does this guide establish induction of SPRED1?
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 SPRED1?
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 A03613 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 SPRED1 localization affect quantitation?
Quantitation · SPRED1 is annotated at the cell membrane, including caveolae, and in the nucleus. Compare equivalent sample preparations and fractions across lanes; a change in one fraction could reflect redistribution rather than a change in total SPRED1.
Should SPRED1 run exactly at its predicted 50.5 kDa?
Interpretation · 50.5 kDa is the predicted mass; no observed band position is supplied. The listed modifications do not establish a visible shift or explain any difference between apparent and predicted mass. Use the predicted mass as a reference, then verify band identity experimentally.
SPRED1 is annotated as a homodimer and as a heterodimer with SPRED2. That interaction alone does not establish that a dimer survives Western-blot sample preparation or produces a particular band. Verify the higher band's identity before assigning it to a dimer.
No signal peptide or propeptide is listed, so the supplied features do not predict a processed form from either. A lower band needs independent identification; these features alone do not establish its origin.
SPRED1 is annotated as a homodimer and as a heterodimer with SPRED2. That interaction alone does not establish that a dimer survives Western-blot sample preparation or produces a particular band. Verify the higher band's identity before assigning it to a dimer.
No signal peptide or propeptide is listed, so the supplied features do not predict a processed form from either. A lower band needs independent identification; these features alone do not establish its origin.
Boster reagents
SPRED1 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-Spred1 Antibody
Cat # A03613
Real WB data
Anti-SPRED1 Antibody Picoband®
Cat # A03613-1
Two the supplier anti-SPRED1 antibodies have WB images: A03613 in mouse brain lysate at 1–2 μg/mL, and A03613-1 in human K562, PC-3, and HEL lysates at 0.5 μg/mL. The shown tests cover these samples and conditions.
Which to pick: For mouse brain WB, consider A03613, whose listed reactivity includes human, mouse, and rat. For human cell lysates, consider A03613-1, which has a WB image showing an approximately 50 kDa band in K562, PC-3, and HEL samples.
Source: BosterBio SPRED1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q7Z699.
- Human Protein Atlas. SPRED1 tissue expression.
- PMC5308578 — target-verified WB comparison
- PMC13229218 — target-verified WB comparison
- PMC13383870 — target-verified WB comparison