UBE2T / Ubiquitin-conjugating enzyme E2 T · Western blot design guide

Design a Western Blot for UBE2T

Source-linked UBE2T Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UBE2T 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 UBE2T: expected band ~22.5 kDa, hero antibody A05874-3, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **UBE2T** Western blot protocol sheet — expected band ~22.5 kDa, antibody A05874-3, controls and PMC citations. Open the full UBE2T WB guide →

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UBE2T 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	~22.5 kDa
	Observed band	~23 kDa
	Gel	10% (catalog A05874-3)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Phosphorylated + Ubl conjugation
	Caveat	Modification-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	1 isoform(s)

Section 1

Source-Linked UBE2T Western Blot Protocol Options

The A05874-3 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	human Hela, human Skov3, human MCF-7, human K562 (catalog A05874-3)
Gel %	10% (catalog A05874-3)
Load	30 ug; reducing conditions (catalog A05874-3)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05874-3)
Membrane	nitrocellulose membrane (catalog A05874-3)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A05874-3)
Primary antibody	A05874-3 · 1:1000 (catalog A05874-3)
Primary incubation	overnight at 4°C (catalog A05874-3)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A05874-3)
Secondary incubation	1.5 hour at RT (catalog A05874-3)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A05874-3)
Detection	ECL (catalog A05874-3)

Section 2

What Is the Expected UBE2T Western Blot Band Size?

UBE2T is predicted at 22.5 kDa and observed near 23 kDa; the cause of the small difference is not established.

What am I looking at on my blot?
Single band near 23 kDa	consistent with the 22.5 kDa predicted UBE2T protein and the observed band
Faint band in a soluble lysate fraction	UBE2T may be retained with nuclear chromatin
Band enriched in a chromatin fraction	consistent with annotated chromatin accumulation
Closely spaced bands near 23 kDa	could reflect different Ser184 phosphorylation states, but band identity requires confirmation

💡Expected UBE2T appearanceUBE2T has a predicted mass of 22.5 kDa and an empirical band near 23 kDa in reducing whole-cell blots; confirm band identity with appropriate controls.

How each factor affects band size
Predicted UBE2T mass	places the unmodified protein near 22.5 kDa
197-residue UBE2T sequence	underlies the predicted 22.5 kDa mass; the observed band is near 23 kDa
Ser184 phosphoserine	may alter mobility, but no visible shift is established
Ser184 phosphorylation state	a mobility difference between states would require experimental confirmation

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	UBE2T may be poorly recovered from the chromatin-associated nuclear fraction	check nuclear and chromatin fractions and verify extraction
Weak or no signal	incomplete recovery of chromatin-associated UBE2T	compare whole-cell and nuclear fractions with extraction controls
Band higher than expected	a larger band is not explained by the supplied features	verify identity using UBE2T depletion and a second antibody
Multiple bands	Ser184 phosphorylation is a possibility, but distinct bands are not established	compare phosphatase-treated samples and confirm bands by UBE2T depletion
Fragments below expected size	the supplied features do not establish a smaller UBE2T product	check sample integrity and confirm fragments with an independent antibody

Source: UniProt Q9NPD8 Human Protein Atlas product datasheet (A05874-3)

Experiment evidence

Western blot images showing UBE2T band

A05874-3 · UBE2T

M05874-1 · UBE2T

← swipe / scroll for more images →

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

Sample controls for UBE2T Western blot

🧪For positive controls for UBE2T in Western blot, you can use a validated UBE2T-expressing cell lysate; no HPA positive tissue or cell 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 data are unavailable, so tissue controls cannot be selected; use knockdown or KO to confirm specificity.

HPA tissue expression evidence for UBE2T

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 UBE2T Western Blot Tips

Deeper troubleshooting and optimisation questions for UBE2T, answered from its protein features.

How should UBE2T 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 UBE2T isoforms explain multiple bands?

Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no basis for assigning additional bands to UBE2T isoforms.

How should Ser184 phosphorylation be assessed?

PTM · UniProt identifies phosphoserine at position 184 in its own numbering. Compare a phospho-specific signal with total UBE2T if assessing this site; do not infer phosphorylation from band position alone. Paper or antibody numbering may differ from UniProt numbering.

Does this guide establish induction of UBE2T?

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 UBE2T Western blot?

Transfer · UBE2T is 197 residues with a predicted mass of 22.5 kDa. Check that your transfer conditions retain and detect the ~23-kDa band. The supplied features do not establish one optimal transfer method.

How should blocking be optimized?

Blocking · Standard workflow guidance: follow the A05874-3 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 the UBE2T band be quantified?

Quantitation · Measure the ~23-kDa band consistently across samples, using signals within the assay’s linear range. For nuclear or chromatin samples, compare equivalent fractions because UniProt reports nuclear localization and chromatin accumulation.

Why is UBE2T observed near 23 kDa instead of 22.5 kDa?

Interpretation · The observed ~23-kDa band is close to UBE2T’s predicted 22.5 kDa. UniProt lists phosphoserine at Ser184, but that feature alone does not establish a visible shift or explain the small difference.

UniProt places UBE2T in the nucleus and notes that it accumulates on chromatin. If whole-cell signal is weak, examine a nuclear or chromatin fraction and compare like fractions across samples.

The record lists one isoform, no signal or propeptide, no glycosylation sites, and phosphoserine at UniProt Ser184. These features do not establish the identity of an extra band. Check antibody specificity before attributing it to UBE2T or its modification.

Boster reagents

UBE2T 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 HSPC150/UBE2T using anti-HSPC150/UBE2T antibody (A05874-3). 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 Hela whole cell lysates, Lane 2: human Skov3 whole cell lysates, Lane 3: human MCF-7 whole cell lysates, Lane 4: human K562 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-HSPC150/UBE2T antigen affinity purified polyclonal antibody (A05874-3) at 1:1000 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 HSPC150/UBE2T at approximately 23 kDa. The expected band size for HSPC150/UBE2T is at 23 kDa.

Anti-HSPC150/UBE2T Antibody Picoband®

Cat # A05874-3

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Real WB data Western blot analysis of HSPC150 expression in HeLa cell lysate.

Anti-HSPC150 Rabbit Monoclonal Antibody

Cat # M05874-1

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Two the supplier antibodies for UBE2T have Western blot images: A05874-3 shows an approximately 23 kDa band in four human cell lines, while M05874-1 shows a blot of HeLa lysate. The supplied evidence does not establish performance in mouse or rat samples.

Which to pick: For human samples, A05874-3 has the more detailed blot caption, including four cell lines and a 1:1000 primary dilution. M05874-1 has a HeLa blot. A05874-3 lists mouse and rat reactivity, but the supplied blot uses human samples only.

Source: BosterBio UBE2T gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9NPD8.
Human Protein Atlas. UBE2T tissue expression.
PMC4924777 — target-verified WB comparison
PMC6750879 — target-verified WB comparison
PMC9899609 — target-verified WB comparison