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- Table of Contents
SESN2 / Sestrin-2 · Western blot design guide
Design a Western Blot for SESN2
Real validated SESN2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SESN2 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
SESN2 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 | ~54.5 kDa | |
| Observed band | ~60 kDa | |
| Gel | 10% (catalog A02558-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated SESN2 Western Blot Protocols
The A02558-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human HEL, human Jurkat (catalog A02558-1) |
| Gel % | 10% (catalog A02558-1) |
| Load | 30 ug; reducing conditions (catalog A02558-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02558-1) |
| Membrane | nitrocellulose membrane (catalog A02558-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02558-1) |
| Primary antibody | A02558-1 · 0.5 μg/mL (catalog A02558-1) |
| Primary incubation | overnight at 4°C (catalog A02558-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02558-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02558-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02558-1) |
| Detection | ECL (catalog A02558-1) |
Section 2
What Is the Expected SESN2 Western Blot Band Size?
SESN2 is predicted at 54.5 kDa and observed at ~60 kDa in reducing whole-cell lysates; the cause of the difference is not established.
| Band at ~60 kDa | Empirical SESN2 signal in reducing whole-cell lysates; confirm identity with controls |
| Band near 54.5 kDa | Consistent with the predicted SESN2 mass; confirm identity with controls |
| Closely spaced bands near the main band | Ser249 phosphorylation is a possible contributor, but a visible shift is unproven |
| Similar ~60 kDa bands across K562, HEL, Jurkat and RT4 lysates | SESN2 was detected in these whole-cell lysates |
💡Expected SESN2 appearanceSESN2 has a predicted mass of 54.5 kDa and an empirical band at ~60 kDa in reducing whole-cell lysates; the difference is unexplained, so confirm band identity with controls.
| Predicted molecular mass | Places the unmodified 480-residue protein near 54.5 kDa |
| 480-residue sequence | Contributes to the predicted 54.5 kDa mass |
| N-acetylmethionine at residue 1 | Adds a small modification; a visible migration change is unproven |
| Phosphoserine at residue 249 | Adds a small modification; a visible migration change is unproven |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SESN2 signal may be below detection in the tested lysate | Compare with a positive-control K562, HEL, Jurkat or RT4 lysate |
| Band higher than expected | The empirical SESN2 band is ~60 kDa versus a predicted 54.5 kDa; the cause is unknown | Check the ~60 kDa region and verify identity with a second antibody or SESN2 depletion |
| Band lower than expected | A lower band has no assigned SESN2 form in the supplied features | Check for degradation and verify identity with SESN2 depletion |
| Multiple bands | Ser249 phosphorylation could contribute, but distinct bands are unproven | Compare phosphatase-treated and untreated samples and verify bands with SESN2 depletion |
| Weak or no signal | SESN2 abundance or assay sensitivity may be limiting | Run a positive-control lysate and check loading, transfer and antibody conditions |
Experiment evidence
Western blot images showing SESN2 band
A02558-1 · SESN2
A02558 · SESN2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SESN2 Western blot
🧪For positive controls for SESN2 in Western blot, you can use no HPA-nominated sample because no positive tissue or cell is provided.
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-based positive and negative controls cannot be established from the supplied evidence.
HPA tissue expression evidence for SESN2
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 SESN2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SESN2, answered from its protein features.
How should SESN2 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 SESN2 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Which SESN2 modifications matter when choosing an antibody?
PTM · UniProt lists N-acetylmethionine at Met1 and phosphoserine at Ser249. For a phospho-specific antibody, check that its stated site maps to UniProt Ser249; paper or antibody numbering may use a different convention.
Does this guide establish induction of SESN2?
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 SESN2 Western blot?
Transfer · Choose transfer conditions suitable for a protein appearing around 60 kDa, then verify transfer with total-protein staining. The supplied features do not specify a unique transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02558-1 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 SESN2 bands be quantified?
Quantitation · Quantify the band near the reported 60 kDa position consistently across samples and normalize to a measured loading reference. If measuring Ser249 phosphorylation, distinguish the phospho-specific signal from total SESN2; the listed modification does not establish its occupancy in your samples.
Why does SESN2 appear near 60 kDa instead of 54.5 kDa?
Interpretation · The reported apparent band is about 60 kDa; the predicted mass is 54.5 kDa. UniProt lists N-acetylmethionine at position 1 and phosphoserine at position 249, but these features alone do not establish the cause of the difference or demonstrate a visible shift.
The supplied record lists no glycosylation sites, signal peptide, propeptide, or alternative sequence to account for extra bands. Check whether each band tracks with SESN2-specific detection before assigning it; the listed modifications alone do not prove a visible shift.
SESN2 interacts with GATOR2, and leucine negatively regulates that interaction. Record leucine conditions when comparing samples, but this interaction feature alone does not predict a change in SESN2 abundance or band position.
The supplied record lists no glycosylation sites, signal peptide, propeptide, or alternative sequence to account for extra bands. Check whether each band tracks with SESN2-specific detection before assigning it; the listed modifications alone do not prove a visible shift.
SESN2 interacts with GATOR2, and leucine negatively regulates that interaction. Record leucine conditions when comparing samples, but this interaction feature alone does not predict a change in SESN2 abundance or band position.
Boster reagents
SESN2 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
Anti-Sestrin 2/SESN2 Antibody Picoband®
Cat # A02558-1
Real WB data
Anti-SESTRIN2 SESN2 Antibody
Cat # A02558
Two anti-SESN2 antibodies have WB images: A02558-1 shows a band near 60 kDa in four human cell lysates (expected 54 kDa); A02558 shows mouse kidney lysate with and without blocking peptide. These examples do not establish performance in every listed species.
Which to pick: For the shown human cell lysate conditions, consider A02558-1. For mouse kidney, A02558 has a WB image with a blocking-peptide comparison and lists Human, Mouse, and Rat reactivity; the supplied image does not show rat samples.
Source: BosterBio SESN2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P58004.
- Human Protein Atlas. SESN2 tissue expression.
- PMC7156099 — target-verified WB comparison
- PMC5115827 — target-verified WB comparison
- PMC4352509 — target-verified WB comparison