SMURF2 / E3 ubiquitin-protein ligase SMURF2 · Western blot design guide

Design a Western Blot for SMURF2

Real validated SMURF2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SMURF2 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

Western blot protocol sheet for SMURF2: expected band ~86.2 kDa, hero antibody A02585, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **SMURF2** Western blot protocol sheet — expected band ~86.2 kDa, antibody A02585, controls and PMC citations. Open the full SMURF2 WB guide →

Navigate to: Protocols Band Controls Antibodies

SMURF2 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	~86.2 kDa
	Gel	8–10% (standard starting point)
	Positive control ⓘ	Testis (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	Ubl conjugation
	Caveat	Modification-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	1 isoform(s)

Section 1

Real Curated SMURF2 Western Blot Protocols

The A02585 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	C2C12 cell lysate (catalog A02585)
Gel %	8–10% (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	A02585 · 1 μg/ml (catalog A02585)
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 SMURF2 Western Blot Band Size?

SMURF2 is predicted at 86.2 kDa; no empirical band is supplied, and its listed features do not demonstrate a migration shift.

What am I looking at on my blot?
Single band near 86.2 kDa in whole-cell lysate	Consistent with predicted full-length SMURF2; confirm identity with controls
Band near 86.2 kDa in a nuclear fraction	Consistent with SMURF2's reported nuclear location
Band near 86.2 kDa in a cytoplasmic fraction	Consistent with SMURF2's reported cytoplasmic location
Band near 86.2 kDa in a membrane fraction	Consistent with SMURF2's reported membrane location

💡Expected SMURF2 appearanceFull-length SMURF2 has a predicted mass of 86.2 kDa, but no empirical band size is supplied; confirm any candidate band with antibody-specific identity controls.

How each factor affects band size
Predicted full-length mass	Sets 86.2 kDa as the sequence-based reference, not a measured band position
Predicted mass in whole-cell lysate	Provides an 86.2 kDa reference for a candidate full-length band
Predicted mass in nuclear and cytoplasmic fractions	Provides the same 86.2 kDa reference in either fraction
Predicted mass in membrane fractions	Provides the same 86.2 kDa reference in this reported location

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	SMURF2 signal may be below detection or lost during extraction	Check antibody performance with a positive-control lysate
Band higher than expected	Identity or migration of the band is unverified	Compare with a positive control and test whether SMURF2 depletion removes it
Band lower than expected	Protein integrity or band identity is unverified	Use fresh lysate with protease inhibitors and test SMURF2 depletion
Multiple bands	Specificity of additional bands is unverified	Compare the bands before and after SMURF2 depletion
Weak or no signal	Abundance or extraction may be insufficient	Check loading and examine nuclear, cytoplasmic, and membrane fractions

Source: UniProt Q9HAU4 Human Protein Atlas product datasheet (A02585)

Experiment evidence

Western blot images showing SMURF2 band

A02585 · SMURF2

A02585-1 · SMURF2

M02585 · SMURF2

RP1102 · SMURF2

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for SMURF2 Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SMURF2 in Western blot, you can use testis tissue, the strongest HPA positive candidate.

Positive control: Testis (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: Testis and adipose tissue provide contrasting controls, though SMURF2 occurs in multiple cell compartments.

HPA tissue expression evidence for SMURF2

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
Testis	spermatogonia cells	High	Protein (IHC)	HPA →
Adrenal gland	glandular cells	Medium	Protein (IHC)	HPA →
Caudate	neuronal cells	Medium	Protein (IHC)	HPA →
Cerebellum	Purkinje cells	Medium	Protein (IHC)	HPA →
Duodenum	glandular cells	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 →
Cervix	squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Epididymis	glandular cells	Not detected	Protein (IHC)	HPA →
Esophagus	squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Liver	cholangiocytes	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SMURF2 Western Blot Tips

Deeper troubleshooting and optimisation questions for SMURF2, answered from its protein features.

Where should the main SMURF2 band appear?

Band shift · The predicted mass is 86.2 kDa. Use it as a reference when identifying a band; no observed band position was supplied, so the apparent mass cannot be confirmed from these features.

Could SMURF2 isoforms explain multiple bands?

Isoforms · The supplied record lists one isoform and no alternative sequence. These features do not support assigning additional bands to specific isoforms.

Do annotated modifications prove a band shift?

PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.

Does this guide establish induction of SMURF2?

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 SMURF2?

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 A02585 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 can SMURF2 localization affect quantitation?

Quantitation · SMURF2 is cytoplasmic in the presence of SMAD7 and colocalizes with CAV1, SMAD7, and the TGF-beta receptor in membrane rafts. Compare like fractions across samples and account for changes in localization when interpreting band intensity.

Does ubiquitin conjugation predict a SMURF2 band shift?

Interpretation · The record includes ubiquitin-like conjugation keywords but lists no modified residues or coordinates. Those keywords alone do not establish a visible shift or explain a difference from 86.2 kDa.

SMURF2 is reported in the nucleus, cytoplasm, cell membrane, and membrane rafts. Check the fractions relevant to your experiment when a whole-cell signal is weak or variable.

Compare them with the 86.2 kDa prediction, but avoid assigning them to an isoform or modification from this record alone. The supplied features give neither an empirical band position nor a modification site that identifies an extra band.

Boster reagents

SMURF2 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 Western blot analysis of SMURF2 in C2C12 cell lysate with SMURF2 antibody at 1 μg/ml.

Anti-SMURF2 Antibody

Cat # A02585

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Real WB data

Western blot analysis of SMURF2 using anti-SMURF2 antibody (A02585-1). Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human HCT116 whole cell lysates, Lane 2: human U2OS whole cell lysates, Lane 3: human SH-SY5Y whole cell lysates, Lane 4: rat testis tissue lysates, Lane 5: rat brain tissue lysates, Lane 6: mouse brain tissue lysates. After electrophoresis, proteins were traSMURF2erred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SMURF2 antigen affinity purified polyclonal antibody (A02585-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for SMURF2 at approximately 86 kDa. The expected band size for SMURF2 is at 86 kDa.

Anti-SMURF2 Antibody Picoband®

Cat # A02585-1

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Real WB data Western blot analysis of SMURF 2 expression in SH-SY-5Y cell lysate.

Anti-SMURF 2 Rabbit Monoclonal Antibody

Cat # M02585

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Real WB data

Western blot analysis of SMURF2 using anti-SMURF2 antibody (RP1102). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: SMMC whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SMURF2 antigen affinity purified polyclonal antibody (Catalog # RP1102) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SMURF2 at approximately 86KD. The expected band size for SMURF2 is at 86KD.

Anti-SMURF 2/SMURF2 Antibody Picoband®

Cat # RP1102

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Four the supplier anti-SMURF2 antibodies list Human, Mouse, and Rat reactivity and have WB images. Captions document specific tested lysates; A02585-1 and RP1102 report bands near the expected 86 kDa. These examples do not establish performance in every sample type.

Which to pick: Choose A02585-1 for the most detailed pictured examples across human cell, rat tissue, and mouse tissue lysates. A02585 shows C2C12 lysate, M02585 shows SH-SY-5Y lysate, and RP1102 shows SMMC lysate. Match your sample and conditions to the closest documented example.

Source: BosterBio SMURF2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9HAU4.
Human Protein Atlas. SMURF2 tissue expression.
PMC5428923 — target-verified WB comparison
PMC7176845 — target-verified WB comparison
PMC6521037 — target-verified WB comparison