FBXO32 / F-box only protein 32 · Western blot design guide

Design a Western Blot for FBXO32

Real validated FBXO32 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FBXO32 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 FBXO32: expected band ~41.6 kDa, hero antibody M02531-1, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **FBXO32** Western blot protocol sheet — expected band ~41.6 kDa, antibody M02531-1, controls and PMC citations. Open the full FBXO32 WB guide →

Navigate to: Protocols Band Controls Antibodies

FBXO32 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	~41.6 kDa
	Observed band	~42 kDa
	Gel	10% (catalog M02531-1)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	—
	Caveat	—
	Gene-set association	MSigDB C7 membership
	Isoform	2 isoform(s)

Section 1

Real Curated FBXO32 Western Blot Protocols

The M02531-1 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	rat heart, rat skeletal muscle, mouse heart (catalog M02531-1)
Gel %	10% (catalog M02531-1)
Load	30 ug; reducing conditions (catalog M02531-1)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02531-1)
Membrane	nitrocellulose membrane (catalog M02531-1)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog M02531-1)
Primary antibody	M02531-1 · 1:500 (catalog M02531-1)
Primary incubation	overnight at 4°C (catalog M02531-1)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog M02531-1)
Secondary incubation	1.5 hour at RT (catalog M02531-1)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog M02531-1)
Detection	ECL (catalog M02531-1)

Section 2

What Is the Expected FBXO32 Western Blot Band Size?

FBXO32 is predicted at 41.6 kDa and observed at ~42 kDa; the cause of the small difference is not established.

What am I looking at on my blot?
Band at ~42 kDa	Consistent with the observed FBXO32 band and its 41.6 kDa predicted mass
Single band near 42 kDa	Isoforms 1 and 2 are not necessarily resolved
Additional band at a different size	Could reflect alternative splicing, but its identity needs confirmation
Different signal between cytoplasmic and nuclear fractions	Consistent with FBXO32 shuttling between these compartments

💡Expected FBXO32 appearanceFBXO32 has a predicted mass of 41.6 kDa and an empirical band at ~42 kDa; confirm band identity with appropriate antibody and sample controls.

How each factor affects band size
Predicted FBXO32 mass	41.6 kDa predicts a band near the observed ~42 kDa
Isoform 1	May contribute to the observed band; its individual migration is unknown
Isoform 2	May migrate differently from isoform 1; its individual migration is unknown
Alternative splicing	Can alter protein size, but distinct bands are not established

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	FBXO32 signal may be below detection in the sampled lysate	Check loading and antibody performance with a known positive lysate
Band higher than expected	The band may be nonspecific or an unverified isoform	Compare with a positive control and verify antibody specificity
Band lower than expected	An alternatively spliced product is possible, but its migration is unknown	Check antibody specificity and compare with the ~42 kDa control band
Multiple bands	Isoforms 1 and 2 are reported, though distinct migration is unproven	Confirm band identity with an independent antibody or FBXO32 depletion
Weak or no signal	FBXO32 distributes between cytoplasm and nucleus	Check a whole-cell lysate and verify fraction loading

Source: UniProt Q969P5 Human Protein Atlas product datasheet (M02531-1)

Experiment evidence

Western blot images showing FBXO32 band

M02531-1 · FBXO32

A02531-1 · FBXO32

← swipe / scroll for more images →

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

Sample controls for FBXO32 Western blot

🧪For positive controls for FBXO32 in Western blot, you can use a verified FBXO32-expressing cell lysate; no HPA positive sample is supplied.

Positive control: No high/medium HPA tissue identified

Negative control: Suggested KO / knockdown lysate

Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.

⚠️Feasibility: HPA provides no tissue data, so tissue controls cannot be selected from the supplied evidence.

HPA tissue expression evidence for FBXO32

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
No high/medium HPA tissues identified in the supplied evidence.

Lower expression tissues · IHC evidence, not confirmed WB-negative controls

Tissue	Cell type	Level	Evidence	Source
No lower-expression tissue rows available in the supplied evidence.

Section 3

Advanced FBXO32 Western Blot Tips

Deeper troubleshooting and optimisation questions for FBXO32, answered from its protein features.

How should FBXO32 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.

How might FBXO32 isoform 2 appear on a blot?

Isoforms · Isoform 2 lacks residues 124–216 relative to canonical isoform 1, so it may run below the ~42 kDa canonical band. The supplied features do not establish its apparent mass or whether it produces a detectable band.

Yes. An antibody recognizing a sequence within canonical residues 124–216 may miss isoform 2, which lacks that region. Check the antibody's stated epitope before interpreting a single band as evidence that only one isoform is present.

Choose the measurement to match the question. Quantify distinguishable bands separately when comparing isoforms 1 and 2. For total FBXO32, first check that the antibody can recognize both, including isoform 2 with residues 124–216 missing.

Do the supplied features support a PTM-related band shift?

PTM · No modified residues or glycosylation sites are listed for FBXO32. Its role in an E3 ubiquitin-protein ligase complex does not itself establish modification of FBXO32 or explain a shifted band.

Does this guide establish induction of FBXO32?

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 FBXO32?

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 M02531-1 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 FBXO32 be quantified across cell fractions?

Quantitation · FBXO32 is reported in both cytoplasm and nucleus and shuttles between them. Quantify the relevant fraction consistently; a change in one fraction could reflect redistribution rather than a change in total FBXO32.

Does the ~42 kDa band match predicted FBXO32 mass?

Interpretation · Yes. The observed ~42 kDa band is close to the 41.6 kDa predicted mass. This agreement supports the assignment but does not establish band identity by itself.

Consider isoform 2 because it lacks canonical residues 124–216. Its presence alone does not identify an observed band. Check whether the antibody recognizes sequence retained in isoform 2 before assigning that band to FBXO32.

Boster reagents

FBXO32 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 FBX32 using anti-FBX32 antibody (M02531-1). 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: rat heart tissue lysates, Lane 2: rat skeletal muscle tissue lysates, Lane 3: mouse heart tissue lysates, Lane 4: mouse skeletal muscle tissue lysates. After electrophoresis, proteins were transferred 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-FBX32 antigen affinity purified monoclonal antibody (M02531-1) at 1:500 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 FBX32 at approximately 42 kDa. The expected band size for FBX32 is at 42 kDa.

Anti-Fbx32 Rabbit Monoclonal Antibody

Cat # M02531-1

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Real WB data

Western blot analysis of Fbx32/FBXO32 using anti-Fbx32/FBXO32 antibody (A02531-1). 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 30 ug of sample under reducing conditions. Lane 1: mouse heart tissue lysates, Lane 2: mouse skeletal muscle tissue lysates. After electrophoresis, proteins were transferred 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-Fbx32/FBXO32 antigen affinity purified polyclonal antibody (Catalog # A02531-1) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Fbx32/FBXO32 at approximately 42 kDa. The expected band size for Fbx32/FBXO32 is at 42 kDa.

Anti-FBXO32 Antibody Picoband®

Cat # A02531-1

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Two the supplier anti-FBXO32 antibodies have Western blot images showing a band near the expected 42 kDa: M02531-1 in rat and mouse heart and skeletal muscle lysates, and A02531-1 in mouse heart and skeletal muscle lysates. Neither supplied image shows human samples.

Which to pick: For rat samples, choose M02531-1; it lists rat reactivity and shows rat tissue blots. For mouse samples, both have relevant blot images. Both list human reactivity, but the supplied images do not demonstrate performance in human samples.

Source: BosterBio FBXO32 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q969P5.
Human Protein Atlas. FBXO32 tissue expression.
PMC12383710 — target-verified WB comparison
PMC12268887 — target-verified WB comparison
PMC8184796 — target-verified WB comparison