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- Table of Contents
HUS1 / Checkpoint protein HUS1 · Western blot design guide
Design a Western Blot for HUS1
Real validated HUS1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HUS1 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
HUS1 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 | ~31.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Colon (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated HUS1 Western Blot Protocols
The A02486 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A02486) |
| 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 | A02486 · 1:1000 (catalog A02486) |
| 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 HUS1 Western Blot Band Size?
HUS1 has a predicted mass of 31.7 kDa; its two isoforms could affect migration, but distinct bands and an empirical size are not demonstrated.
| Band near 31.7 kDa | Consistent with the predicted HUS1 mass, pending identity controls |
| Several bands at different positions | HUS1 has isoforms 1 and 2, though their separation is unestablished |
| Band in a nuclear fraction | Consistent with nuclear HUS1 localization |
| Stronger nuclear signal after DNA damage | Consistent with HUS1 accumulation in nuclear foci |
💡Expected HUS1 appearanceHUS1 has a predicted mass of 31.7 kDa; no empirical band size or isoform migration is supplied, so confirm any candidate band with ordinary identity controls.
| Predicted HUS1 mass | 31.7 kDa provides a reference for the sequence-based band position |
| Isoform 1 | Its apparent size relative to isoform 2 is unknown |
| Isoform 2 | Its apparent size relative to isoform 1 is unknown |
| Alternative splicing | May affect band position, but distinct migration is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | HUS1 may be diluted across nuclear and cytosolic compartments | Check nuclear and cytosolic fractions and verify antibody performance |
| Band higher than expected | The supplied features do not establish a size-increasing modification | Confirm identity with an independent antibody or HUS1 depletion |
| Band lower than expected | Isoform migration is unknown and the lower species may be unrelated | Confirm identity with HUS1 depletion or an independent antibody |
| Multiple bands | HUS1 has two splice isoforms, but their band positions are unknown | Use isoform-specific evidence and HUS1 depletion to assign bands |
| Weak or no signal | HUS1 distribution between cytosol and nucleus can change after DNA damage | Compare matched damage conditions and examine nuclear fractions |
Experiment evidence
Western blot images showing HUS1 band
A02486 · HUS1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HUS1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HUS1 in Western blot, you can use adipose tissue lysate, where HPA reports medium expression.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Colon (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA identifies a not-detected tissue for comparison, but the positive tissue shows only medium expression.
HPA tissue expression evidence for HUS1
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Colon | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HUS1 Western Blot Tips
Deeper troubleshooting and optimisation questions for HUS1, answered from its protein features.
Where should the main HUS1 band migrate?
Band shift · The predicted mass of canonical HUS1 is 31.7 kDa. No empirical apparent band is supplied, so use 31.7 kDa as a starting estimate, not a confirmed migration position.
Could HUS1 isoforms produce different bands?
Isoforms · UniProt lists two isoforms. Isoform 2 lacks residues 1–21 of the canonical sequence, so it may migrate below isoform 1. This sequence difference does not establish that two bands will be visible.
Should a slower band be assigned to a HUS1 modification?
PTM · UniProt lists no modified residues or glycosylation sites for HUS1. A slower band alone does not identify a modification or explain a difference from the predicted 31.7 kDa mass.
Does this guide establish induction of HUS1?
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 HUS1?
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 A02486 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 HUS1 be quantified across cellular fractions?
Quantitation · HUS1 is reported in both the nucleus and cytosol, with nuclear foci after DNA damage. Quantify comparable fractions separately and report which fraction was measured; a change in one fraction need not reflect total cellular HUS1.
How might DNA damage affect a HUS1 blot?
Interpretation · HUS1 appears in discrete nuclear foci upon DNA damage. Compare matched conditions and consider nuclear and cytosolic fractions when assessing a change; the supplied features do not establish an increase in total HUS1 abundance.
HUS1 is a component of the RAD9A–RAD1–HUS1 complex. Complex membership alone does not establish that an intact complex will appear as a higher band on a Western blot.
Consider isoform 2, which lacks canonical residues 1–21, as one possibility. The supplied features do not give its apparent mass or confirm that an observed lower band is isoform 2.
HUS1 is a component of the RAD9A–RAD1–HUS1 complex. Complex membership alone does not establish that an intact complex will appear as a higher band on a Western blot.
Consider isoform 2, which lacks canonical residues 1–21, as one possibility. The supplied features do not give its apparent mass or confirm that an observed lower band is isoform 2.
Boster reagents
HUS1 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-Checkpoint protein HUS1 HUS1 Antibody
Cat # A02486
The catalog reports anti-HUS1 antibody A02486 for Western blotting, with reported human and mouse reactivity. Its WB image shows extracts from various cell lines tested at a 1:1000 antibody dilution; the supplied caption does not identify the cell lines.
Which to pick: A02486 is the only listed HUS1 option and has a WB image. Check its reported human and mouse reactivity against your sample; the image caption does not establish which species were tested.
Source: BosterBio HUS1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O60921.
- Human Protein Atlas. HUS1 tissue expression.
- PMC9031773 — target-verified WB comparison
- PMC6231531 — target-verified WB comparison