This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
HES1 / Transcription factor HES-1 · Western blot design guide
Design a Western Blot for HES1
Real validated HES1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HES1 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
HES1 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 | ~29.5 kDa | |
| Observed band | ~30 kDa | |
| Gel | 5–20% (catalog M01459) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated HES1 Western Blot Protocols
The M01459 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human U-87MG, human U251, human 293T (catalog M01459) |
| Gel % | 5–20% (catalog M01459) |
| Load | 30 ug; reducing conditions (catalog M01459) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01459) |
| Membrane | nitrocellulose membrane (catalog M01459) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01459) |
| Primary antibody | M01459 · 1:500 (catalog M01459) |
| Primary incubation | overnight at 4°C (catalog M01459) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M01459) |
| Secondary incubation | 1.5 hour at RT (catalog M01459) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01459) |
| Detection | ECL (catalog M01459) |
Section 2
What Is the Expected HES1 Western Blot Band Size?
HES1 is predicted at 29.5 kDa and observed at ~30 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~30 kDa | Matches the empirical HES1 band and its 29.5 kDa predicted mass |
| Single sharp band near 30 kDa | Consistent with the single annotated HES1 isoform |
| Band well above 30 kDa | No supplied HES1 feature establishes a higher migrating species |
| Band well below 30 kDa | No signal peptide or propeptide cleavage is annotated for HES1 |
💡Expected HES1 appearanceHES1 has a predicted mass of 29.5 kDa and an empirical band at ~30 kDa; confirm band identity with a validated positive control and antibody specificity controls.
| UniProt predicted mass | Places the expected HES1 band near 29.5 kDa |
| 280 amino acid sequence | Underlies the predicted mass without establishing a separate processed band |
| Single annotated isoform | Provides no basis for assigning distinct isoform band sizes |
| No annotated glycosylation sites | Provides no basis for attributing a size shift to glycosylation |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear HES1 may be poorly represented in the prepared lysate | Check nuclear extraction and run a validated positive control |
| Band higher than expected | No supplied HES1 feature establishes a higher migrating species | Check antibody specificity and compare with a positive control |
| Band lower than expected | No annotated signal peptide or propeptide explains a smaller band | Check sample integrity and antibody specificity |
| Multiple bands | Only one HES1 isoform is annotated here | Use a specificity control to identify the HES1 band |
| Weak or no signal | Nuclear HES1 may be underrepresented in the sample | Check nuclear protein recovery and antibody performance with a positive control |
Experiment evidence
Western blot images showing HES1 band
M01459 · HES1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HES1 Western blot
🧪For positive controls for HES1 in Western blot, you can use an HPA-IHC candidate positive sample once one is identified.
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: HES1 is nuclear, but the supplied HPA data cannot establish a positive or negative sample.
HPA tissue expression evidence for HES1
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 HES1 Western Blot Tips
Deeper troubleshooting and optimisation questions for HES1, answered from its protein features.
How should HES1 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 HES1 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. Check whether an additional band is reproducible before assigning it to HES1.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of HES1?
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 HES1 Western blot?
Transfer · HES1 is 280 amino acids with a predicted mass of 29.5 kDa. Choose and verify transfer conditions that retain a protein near 30 kDa; the supplied features do not establish a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01459 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 HES1 bands be quantified?
Quantitation · Quantify the reproducible band near 30 kDa using the same band boundary across samples. HES1 is annotated as nuclear, so keep sample fractions consistent when comparing signals. Do not include additional bands in the measurement without evidence that they represent HES1.
Should HES1 run above its predicted mass?
Interpretation · HES1 has a predicted mass of 29.5 kDa, close to the observed band near 30 kDa. This agreement does not require a modification-based explanation. Compare bands against a molecular-weight marker and evaluate larger deviations separately.
Ubl conjugation is listed as a keyword, but no modified residue or position is supplied. The keyword alone does not establish a visible shift or identify a particular shifted band. Interpret any extra band using independent evidence.
Start with the expected band near 30 kDa. HES1 interacts with TLE1, more weakly with TLE2, and with HES6, but those interactions do not identify an unexpected Western-blot band. The record also gives no alternative isoform sequence or mapped modification that assigns one.
Ubl conjugation is listed as a keyword, but no modified residue or position is supplied. The keyword alone does not establish a visible shift or identify a particular shifted band. Interpret any extra band using independent evidence.
Start with the expected band near 30 kDa. HES1 interacts with TLE1, more weakly with TLE2, and with HES6, but those interactions do not identify an unexpected Western-blot band. The record also gives no alternative isoform sequence or mapped modification that assigns one.
Boster reagents
HES1 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-Hes1 Rabbit Monoclonal Antibody
Cat # M01459
M01459 is a rabbit monoclonal anti-HES1 antibody with a reported Western blot band near 30 kDa in human cell lysates and rat and mouse brain lysates. This evidence covers the specimens and conditions shown in the product image.
Which to pick: M01459 is the only listed option. Its Western blot image includes human U-87MG, U251, and 293T cells plus rat and mouse brain lysates, tested at 1:500 under reducing conditions. Choose it when those reported contexts fit your experiment.
Source: BosterBio HES1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14469.
- Human Protein Atlas. HES1 tissue expression.
- PMC3572096 — target-verified WB comparison
- PMC5858497 — target-verified WB comparison
- PMC4742206 — target-verified WB comparison