APOBEC3B / DNA dC->dU-editing enzyme APOBEC-3B · Western blot design guide

Design a Western Blot for APOBEC3B

Source-linked APOBEC3B Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-APOBEC3B 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 APOBEC3B: expected band ~45.9 kDa, hero antibody A01088, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **APOBEC3B** Western blot protocol sheet — expected band ~45.9 kDa, antibody A01088, controls and PMC citations. Open the full APOBEC3B WB guide →

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APOBEC3B 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	~45.9 kDa
	Gel	12–15% (standard starting point)
	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	3 isoform(s)

Section 1

Source-Linked APOBEC3B Western Blot Protocol Options

The A01088 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	HepG2 cell lysate (catalog A01088)
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	A01088 · 1 μg/ml (catalog A01088)
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 APOBEC3B Western Blot Band Size?

APOBEC3B has a predicted monomer mass of 45.9 kDa; homodimerization and three isoforms could affect bands, but distinct migration is unverified.

What am I looking at on my blot?
Band near 45.9 kDa	Consistent with the predicted monomer; confirm identity with appropriate controls
Higher band near 92 kDa	Could reflect a homodimer retained during electrophoresis
Several bands at different positions	Could reflect isoforms 1, 2, and 3; their migration differences are unknown
Weak band in whole-cell lysate	Nuclear localization may make the signal clearer in a nuclear fraction

💡Expected APOBEC3B appearanceThe predicted monomer is 45.9 kDa; no empirical band size is supplied, so confirm any band near that position with band-identity controls.

How each factor affects band size
Predicted monomer mass	Provides a 45.9 kDa reference, not a measured migration position
Homodimer formation	Could produce a band near twice the monomer mass if the dimer remains intact during electrophoresis
Isoform 1	Apparent size relative to the other isoforms is unknown
Isoform 2	May migrate differently from other isoforms; its size is unspecified
Isoform 3	May migrate differently from other isoforms; its size is unspecified

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	Nuclear APOBEC3B may be insufficiently recovered or detected	Check nuclear fraction recovery and use a positive control
Band higher than expected	A homodimer may remain associated	Compare thoroughly denatured samples and confirm band identity
Band lower than expected	An isoform may differ in size, but its migration is unknown	Confirm identity with an independent antibody or genetic control
Multiple bands	Isoforms 1, 2, and 3 may contribute, but distinct bands are unverified	Compare isoform-specific expression or use a genetic control
Weak or no signal	The nuclear target may be dilute in whole-cell lysate	Test a nuclear fraction alongside a positive control

Source: UniProt Q9UH17 Human Protein Atlas product datasheet (A01088)

Experiment evidence

Western blot images showing APOBEC3B band

A01088 · APOBEC3B

A01088-1 · APOBEC3B

← swipe / scroll for more images →

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

Sample controls for APOBEC3B Western blot

🧪For positive controls for APOBEC3B in Western blot, you can use an HPA-IHC candidate positive sample once one is identified; none is supplied here.

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 expression data are absent, so tissue controls cannot be established from this evidence; use a knockdown or KO for a negative control.

HPA tissue expression evidence for APOBEC3B

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 APOBEC3B Western Blot Tips

Deeper troubleshooting and optimisation questions for APOBEC3B, answered from its protein features.

How should APOBEC3B 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 could APOBEC3B isoforms affect band interpretation?

Isoforms · Three isoforms are listed. Relative to the canonical sequence, isoform 2 replaces residues 191–382 with a shorter sequence, and isoform 3 lacks residues 242–266. Either sequence change could affect apparent band position; the features do not establish where either isoform runs.

An epitope within canonical residues 191–382 may be changed in isoform 2; residues 242–266 are also absent from isoform 3. Check the antibody’s mapped epitope against all three sequences before interpreting a missing isoform band.

Decide whether the measurement covers all isoforms or a specific isoform. Isoform 2 changes canonical residues 191–382, while isoform 3 lacks residues 242–266; an antibody recognizing either region may detect the isoforms differently. Compare signals only after checking epitope coverage.

Do listed modifications explain an unexpected APOBEC3B band?

PTM · The supplied features list no modified residues or glycosylation sites. They therefore provide no specific modification to assign to an unexpected band, and feature presence or absence alone cannot establish its cause. Check antibody specificity and the listed isoform sequences before assigning it.

Does this guide establish induction of APOBEC3B?

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 APOBEC3B?

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 A01088 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 APOBEC3B 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.

Should APOBEC3B run at exactly 45.9 kDa?

Interpretation · 45.9 kDa is the predicted mass of the 382-residue canonical sequence. No observed band position is supplied, so use it as a reference rather than an exact migration target.

UniProt places APOBEC3B in the nucleus. Include a nuclear fraction when choosing samples, and keep the fraction consistent across lanes used for comparison.

APOBEC3B is annotated as a homodimer and interacts with APOBEC3G. Those interactions alone do not identify a higher Western-blot band. Confirm a band’s identity before assigning it to a dimer or complex.

Boster reagents

APOBEC3B 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 APOBEC3B in HepG2 cell lysate with APOBEC3B antibody at 1 μg/ml.

Anti-APOBEC3B Antibody

Cat # A01088

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Real WB data

Western blot analysis of APOBEC3B using anti-APOBEC3B antibody (A01088-1). 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: human HepG2 whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human THP-1 whole cell lysates, Lane 4: human HEK293 whole cell 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-APOBEC3B antigen affinity purified polyclonal antibody (Catalog # A01088-1) 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 APOBEC3B at approximately 35 kDa. The expected band size for APOBEC3B is at 46 kDa.

Anti-APOBEC3B Antibody Picoband®

Cat # A01088-1

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Both listed anti-APOBEC3B antibodies are catalogued for human samples and have Western blot images. A01088 was shown with HepG2 lysate; A01088-1 was shown with four human cell lysates. For A01088-1, the reported band is approximately 35 kDa versus an expected 46 kDa.

Which to pick: For HepG2 lysate, either antibody has a reported blot: A01088 at 1 μg/mL or A01088-1 at 0.5 μg/mL. Choose A01088-1 if the reported Jurkat, THP-1, or HEK293 examples are useful, while accounting for its band-size discrepancy.

Source: BosterBio APOBEC3B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9UH17.
Human Protein Atlas. APOBEC3B tissue expression.
PMC10302536 — target-verified WB comparison
PMC10835258 — target-verified WB comparison
PMC10562255 — target-verified WB comparison