SESN1 / Sestrin-1 · Western blot design guide

Design a Western Blot for SESN1

Source-linked SESN1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SESN1 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 SESN1: expected band ~56.6 kDa, hero antibody A05559-2, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **SESN1** Western blot protocol sheet — expected band ~56.6 kDa, antibody A05559-2, controls and PMC citations. Open the full SESN1 WB guide →

Navigate to: Protocols Band Controls Antibodies

SESN1 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	~56.6 kDa
	Observed band	~56 kDa
	Gel	5–20% (catalog A05559-2)
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Phosphorylated
	Caveat	Phosphorylation-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	3 isoform(s)

Section 1

Source-Linked SESN1 Western Blot Protocol Options

The A05559-2 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 293T, human K562, rat heart, mouse heart (catalog A05559-2)
Gel %	5–20% (catalog A05559-2)
Load	30 ug; reducing conditions (catalog A05559-2)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05559-2)
Membrane	nitrocellulose membrane (catalog A05559-2)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A05559-2)
Primary antibody	A05559-2 · 0.5 μg/mL (catalog A05559-2)
Primary incubation	overnight at 4°C (catalog A05559-2)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A05559-2)
Secondary incubation	1.5 hour at RT (catalog A05559-2)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A05559-2)
Detection	ECL (catalog A05559-2)

Section 2

What Is the Expected SESN1 Western Blot Band Size?

SESN1 is predicted at 56.6 kDa and observed at ~56 kDa; the cause of the small difference is not established.

What am I looking at on my blot?
Band at ~56 kDa	Empirical SESN1 band in reducing whole-cell lysates
Single band near 56.6 kDa	Consistent with the predicted SESN1 mass
Several discrete bands	Could reflect T2, T1, or T3, but their migration is unverified
Close doublet near 56 kDa	Could reflect phosphorylation at Ser293 or Ser314; a mobility shift is unproven

💡Expected SESN1 appearanceSESN1 has a predicted mass of 56.6 kDa and an observed band at ~56 kDa; confirm band identity with an appropriate positive control and SESN1 depletion.

How each factor affects band size
Predicted SESN1 mass	56.6 kDa predicts a band near the observed ~56 kDa
T2 splice isoform	May differ in apparent size; its mass and migration are unspecified
T1 splice isoform	May differ in apparent size; its mass and migration are unspecified
T3 splice isoform	May differ in apparent size; its mass and migration are unspecified

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	SESN1 expression or recovery from nuclear and cytoplasmic material may be insufficient	Check a positive lysate and verify nuclear and cytoplasmic extraction
Band higher than expected	An isoform or nonspecific signal is possible; its identity is unverified	Compare with SESN1-depleted lysate
Band lower than expected	An isoform or nonspecific signal is possible; its identity is unverified	Compare with SESN1-depleted lysate
Multiple bands	T2, T1, and T3 exist, but distinct band sizes are unverified	Use SESN1 depletion and isoform-aware controls to identify the bands
Weak or no signal	SESN1 abundance or recovery may be low	Check loading, transfer, and a positive lysate

Source: UniProt Q9Y6P5 Human Protein Atlas product datasheet (A05559-2)

Experiment evidence

Western blot images showing SESN1 band

A05559-2 · SESN1

M05559 · SESN1

A05559-1 · SESN1

← swipe / scroll for more images →

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

Sample controls for SESN1 Western blot

🧪For positive controls for SESN1 in Western blot, you can use no HPA-supported tissue or cell line because no expression data were 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: Without HPA expression data, a positive sample cannot be selected and a tissue negative cannot be verified.

HPA tissue expression evidence for SESN1

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 SESN1 Western Blot Tips

Deeper troubleshooting and optimisation questions for SESN1, answered from its protein features.

How should SESN1 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 SESN1 isoforms produce different bands?

Isoforms · Three isoforms are listed: T1, T2 and T3. T3 lacks residues 1–66, while T1 has an alternative sequence at residues 1–34. Compare bands with the expected sequences and the antibody’s binding region; the supplied features do not specify each isoform’s apparent mass.

Check whether its binding region overlaps the N-terminal sequences that differ among isoforms. T3 lacks residues 1–66, and T1 replaces the sequence at residues 1–34. An antibody directed at a differing region may not detect every isoform equally.

Where are the annotated SESN1 phosphorylation sites?

PTM · UniProt lists phosphoserine at positions 293 and 314 in its sequence numbering. Check which numbering convention an antibody or paper uses before comparing sites. These annotations alone do not predict a resolvable band shift.

Does this guide establish induction of SESN1?

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 SESN1?

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 A05559-2 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 SESN1 bands be quantified across samples?

Quantitation · Quantify the same band or band set consistently across samples. SESN1 has three annotated isoforms, so state whether the measurement includes one band or several. Keep sample preparation consistent because SESN1 is annotated in both the nucleus and cytoplasm.

Why is the SESN1 band near 56 kDa?

Interpretation · The observed band near 56 kDa is close to the 56.6 kDa predicted mass. Phosphorylation and alternative sequences are annotated, but their presence alone does not establish a visible shift or explain a mass difference.

SESN1 is annotated in both the nucleus and cytoplasm. Keep the fraction or whole-cell preparation consistent across samples, and interpret a change in one fraction separately from a change in total SESN1.

Consider the annotated T1 and T3 N-terminal sequence differences and check whether the antibody can recognize them. Phosphoserines at UniProt positions 293 and 314 are also annotated, but neither these sites nor the isoform records establish the identity of an unexpected band on their own.

Boster reagents

SESN1 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 SESN1 using anti-SESN1 antibody (A05559-2). 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 293T whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: rat heart tissue lysates, Lane 4: mouse heart 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-SESN1 antigen affinity purified polyclonal antibody (Catalog # A05559-2) 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 SESN1 at approximately 56 kDa. The expected band size for SESN1 is at 56,65-75 kDa.

Anti-SESN1 Antibody Picoband®

Cat # A05559-2

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Real WB data

Western blot analysis of SESN1 using anti-SESN1 antibody (M05559). 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 K562 whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human HepG2 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-SESN1 antigen affinity purified monoclonal antibody (Catalog # M05559) 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 1:500 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 SESN1 at approximately 41 kDa. The expected band size for SESN1 is at 47 kDa.

Anti-SESN1 Rabbit Monoclonal Antibody

Cat # M05559

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Real WB data Western blot analysis of lysates from rat muscle cells, using SESN1 Antibody. The lane on the right is blocked with the synthesized peptide.

Western blot analysis of lysates from rat muscle cells, using SESN1 Antibody. The lane on the right is blocked with the synthesized peptide.

Anti-SESN1/Pa26 Antibody

Cat # A05559-1

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Three anti-SESN1 antibodies list human, mouse and rat reactivity and have WB images. Captions show distinct tested lysates; these examples do not establish performance in every tissue or species. Reported bands differ between A05559-2 and M05559.

Which to pick: Pick A05559-2 for pictured human cell, rat heart and mouse heart lysates; M05559 for pictured human cell lysates; or A05559-1 for a rat muscle example with peptide blocking. Check the reported band sizes and conditions against your experiment.

Source: BosterBio SESN1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q9Y6P5.
Human Protein Atlas. SESN1 tissue expression.
PMC8941522 — target-verified WB comparison
PMC10292895 — target-verified WB comparison
PMC12926326 — target-verified WB comparison