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- Table of Contents
HSPE1 / 10 kDa heat shock protein, mitochondrial · Western blot design guide
Design a Western Blot for HSPE1
Source-linked HSPE1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HSPE1 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
HSPE1 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 | ~10.9 kDa | |
| Observed band | ~11 and 13 kDa | |
| Gel | 5–20% (catalog M06192-1) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| 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 Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked HSPE1 Western Blot Protocol Options
The M06192-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A431, human A549, rat heart, rat skeletal muscle, mouse heart, mouse skeletal muscle (catalog M06192-1) |
| Gel % | 5–20% (catalog M06192-1) |
| Load | 30 ug; reducing conditions (catalog M06192-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M06192-1) |
| Membrane | nitrocellulose membrane (catalog M06192-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M06192-1) |
| Primary antibody | M06192-1 · 1:1000 (catalog M06192-1) |
| Primary incubation | overnight at 4°C (catalog M06192-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:1000 (catalog M06192-1) |
| Secondary incubation | 1.5 hour at RT (catalog M06192-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M06192-1) |
| Detection | ECL (catalog M06192-1) |
Section 2
What Is the Expected HSPE1 Western Blot Band Size?
HSPE1 is predicted at 10.9 kDa, while blots show bands near 11 and 13 kDa; the cause of the 13 kDa migration is not established.
| Band near 11 kDa | Matches the predicted 10.9 kDa mass and an observed HSPE1 band |
| Band near 13 kDa | Observed HSPE1 signal with no established cause for its migration |
| Bands near 11 and 13 kDa | Empirical pattern whose two positions are not assigned to distinct isoforms |
| Weak signal in a mitochondrial-poor sample | May reflect HSPE1 localization to the mitochondrial matrix |
💡Expected HSPE1 appearanceHSPE1 has a predicted mass of 10.9 kDa; antibody blots show bands near 11 and 13 kDa, whose identity and differing migration should be checked with ordinary band-identity controls.
| Predicted 10.9 kDa mass | Provides the reference size for the band near 11 kDa |
| N-acetylalanine at residue 2 | Is a documented modification without a demonstrated resolvable band shift |
| N6-acetyllysine at residue 8 | Is a documented modification without a demonstrated resolvable band shift |
| N6-succinyllysine at residue 28 | Is a documented modification without a demonstrated resolvable band shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of mitochondrial matrix protein | Check mitochondrial protein recovery and sample loading |
| Band higher than expected | The observed 13 kDa band's migration is unexplained by the supplied features | Compare with the 11 kDa band and verify identity with an independent antibody or HSPE1 depletion |
| Multiple bands | The observed 11 and 13 kDa bands have no established assignment to separate isoforms | Verify each band with HSPE1 depletion or an independent antibody |
| Weak or no signal | Low mitochondrial protein recovery or insufficient sample | Check mitochondrial recovery and loading |
| Fragments below expected size | Sample degradation is possible but not established by the supplied features | Check sample integrity and compare with a freshly prepared lysate |
Experiment evidence
Western blot images showing HSPE1 band
M06192-1 · HSPE1
PA1790 · HSPE1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HSPE1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HSPE1 in Western blot, you can use adipose tissue lysate, which has high HPA expression.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No clean negative tissue is listed; use siRNA knockdown or a KO line.
HPA tissue expression evidence for HSPE1
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 | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Ovary | follicle cells | Medium | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Medium | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced HSPE1 Western Blot Tips
Deeper troubleshooting and optimisation questions for HSPE1, answered from its protein features.
How should HSPE1 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 an alternative isoform explain two bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for the ~11 and ~13 kDa bands.
Which HSPE1 modifications should guide band interpretation?
PTM · UniProt lists N-acetylalanine at position 2; lysine acetylation at 8 and 99; succinylation at 28; alternate acetylation, malonylation, or succinylation at 40 and 56; alternate malonylation or succinylation at 54; alternate acetylation or succinylation at 66, 70, 80, and 86; and phosphothreonine at 79. These are UniProt coordinates and may differ from antibody or paper numbering. They do not establish a visible band shift.
UniProt lists phosphothreonine at position 79. That site alone does not show that phosphorylation produces the ~13 kDa band. Compare the bands with a suitable phosphatase-treated sample if testing this explanation, and keep UniProt numbering explicit.
UniProt lists phosphothreonine at position 79. That site alone does not show that phosphorylation produces the ~13 kDa band. Compare the bands with a suitable phosphatase-treated sample if testing this explanation, and keep UniProt numbering explicit.
Does this guide establish induction of HSPE1?
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 HSPE1 Western blot?
Transfer · HSPE1 is a small, 102-residue protein with a predicted mass of 10.9 kDa. Choose and verify a transfer setup that retains proteins around 11 kDa, including a membrane suited to small proteins; check transfer and retention before interpreting a missing band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M06192-1 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 HSPE1 be quantified across samples?
Quantitation · HSPE1 is listed in the mitochondrial matrix. Compare equivalent sample fractions and loading across conditions. If both ~11 and ~13 kDa bands appear, quantify them consistently and report which band or bands were measured; the supplied features do not establish that both represent HSPE1.
How should the 11 and 13 kDa bands compare with predicted mass?
Interpretation · HSPE1 is predicted at 10.9 kDa, close to the observed ~11 kDa band; ~13 kDa is higher. Its listed modifications do not establish the cause of that difference or prove a visible shift. Evaluate both bands separately rather than assigning either by size alone.
UniProt describes HSPE1 as a homoheptamer that interacts with Hsp60 assemblies. This describes its protein complex, but does not establish the identity of an unexpected Western-blot band. Assess unexpected bands against the predicted 10.9 kDa mass and the assay conditions.
The supplied record predicts 10.9 kDa and lists one isoform. It also lists modifications, but none is shown here to produce a ~13 kDa band. Treat the observed ~13 kDa signal as unassigned until its identity is independently checked.
UniProt describes HSPE1 as a homoheptamer that interacts with Hsp60 assemblies. This describes its protein complex, but does not establish the identity of an unexpected Western-blot band. Assess unexpected bands against the predicted 10.9 kDa mass and the assay conditions.
The supplied record predicts 10.9 kDa and lists one isoform. It also lists modifications, but none is shown here to produce a ~13 kDa band. Treat the observed ~13 kDa signal as unassigned until its identity is independently checked.
Boster reagents
HSPE1 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-HSP10 Rabbit Monoclonal Antibody
Cat # M06192-1
Real WB data
Anti-Cpn10/HSPE1 Antibody Picoband®
Cat # PA1790
Two the supplier anti-HSPE1 antibodies have Western blot images using human cell lysates and rat and mouse tissues. Both captions report a band near the expected 11 kDa; these examples document the tested samples and do not establish performance in every tissue.
Which to pick: Both M06192-1 and PA1790 list human, mouse, and rat reactivity and show WB images. Choose M06192-1 for a documented skeletal muscle example; PA1790 documents heart tissue and human cell lysates. Their reported primary antibody conditions differ.
Source: BosterBio HSPE1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P61604.
- Human Protein Atlas. HSPE1 tissue expression.
- PMC3650054 — target-verified WB comparison
- PMC12973555 — target-verified WB comparison
- PMC13382145 — target-verified WB comparison