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HSPE1 / 10 kDa heat shock protein, mitochondrial · IHC design guide
Design Immunohistochemistry for HSPE1
Plan HSPE1 paraffin IHC around granular cytoplasmic tissue staining (HPA tissue IHC) and its mitochondrial matrix location (UniProt). This guide covers fixation, staining interpretation and the catalog antibody’s documented IHC workflow (datasheet M06192-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HSPE1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Granular cytoplasm (HPA tissue IHC); mitochondrial matrix (UniProt) | |
| Staining pattern | Granular cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M06192-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M06192-1) | |
| Caveat | HPA staining may reflect proteins from multiple genes (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans 2–102 (UniProt) |
Section 1
Recommended HSPE1 IHC & IF Protocols
The catalog antibody protocol and three published HSPE1 IHC protocols provide starting conditions for paraffin sections (datasheet: IHC-P; PMC6089400; PMC3650054; PMC3964225).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M06192-1) |
| Fixation | Image fixative and duration unreported (datasheet M06192-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet M06192-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M06192-1) |
| Primary antibody | Rabbit monoclonal (clone 19H75) anti-HSPE1, 1:50 (datasheet M06192-1) |
| Primary incubation | Overnight at 4 °C (datasheet M06192-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M06192-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HSPE1-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: M06192-1); published antibody conditions may differ (PMC6089400; PMC3650054; PMC3964225).
Section 2
What Is the Expected HSPE1 Staining Pattern?
HSPE1 is a mitochondrial matrix co-chaperonin with no transmembrane segment (UniProt P61604). In tissue IHC, expect granular cytoplasmic staining, including high staining in adrenal glandular cells, adipocytes and bone marrow hematopoietic cells (HPA tissue IHC). HPA rates the tissue pattern Supported, with medium staining–RNA consistency and a caution that the antibodies target protein from more than one gene (HPA tissue IHC).
| Granular cytoplasmic staining in adrenal glandular cells, adipocytes or bone marrow hematopoietic cells. | This fits the reported high staining in these cells (HPA tissue IHC) and the mitochondrial matrix location (UniProt P61604). Judge the pattern in the identified cell population. |
| Predominantly nuclear or membrane staining, with little granular cytoplasmic signal. | This conflicts with the reported cytoplasmic pattern (HPA tissue IHC) and matrix location without a transmembrane segment (UniProt P61604). Review morphology, detection and antibody specificity before scoring it as HSPE1. |
| Strong staining in an unexpected cell population or in tissue components outside cells. | The supplied HPA profile gives no negative cell populations, so this alone cannot establish a false positive (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity using appropriate IHC controls (standard IHC practice). |
| Uniform haze across cells, stroma or the whole section obscures cytoplasmic granules. | A broad haze is difficult to reconcile with the reported granular cytoplasmic pattern (HPA tissue IHC). Assess background with a primary-omission control and review blocking, washes and detection conditions (standard IHC practice). |
| No stain in adrenal glandular cells, adipocytes or bone marrow hematopoietic cells. | These are reported as high-staining populations (HPA tissue IHC), so first check the run and tissue integrity. A negative section alone does not disprove expression; HPA reports medium staining–RNA consistency (HPA tissue IHC). |
💡Expected HSPE1 appearanceA convincing positive shows granular cytoplasmic staining in an identified high-staining population, such as adrenal glandular cells or adipocytes (HPA tissue IHC); dominant nuclear or membrane staining calls for investigation against the mitochondrial matrix assignment (UniProt P61604).
| Compartment and topology | HSPE1 is assigned to the mitochondrial matrix and has no transmembrane segment (UniProt P61604). Interpret cytoplasmic granules in that context; bright surface staining lacks support from this record. |
| Tissue profile and confidence | HPA describes general granular cytoplasmic expression and lists several high-staining cell populations, but no negative or low controls in this payload (HPA tissue IHC). Its Supported rating includes medium staining–RNA consistency and a multi-gene targeting caution (HPA tissue IHC). |
| Protein forms | The record describes a 102-aa precursor, a chain spanning residues 2–102, and no annotated isoforms or glycosylation sites (UniProt P61604). Those annotations do not establish antibody epitope access or an IHC retrieval condition. |
| IHC versus ICC-IF evidence | Three listed antibodies have Supported IHC status and no stated ICC status (HPA antibodies). HPA supplies no main ICC-IF location or cell-line images (HPA subcellular); do not treat the IHC pattern as an observed IF result. |
| Situation | Likely cause | Next action |
|---|---|---|
| Granular staining is faint in a reported high-staining cell population. | The run may lack adequate signal; this record supplies no HSPE1-specific retrieval or dilution setting (HPA tissue IHC). | Confirm section quality and controls, then optimize retrieval and antibody dilution within the antibody's validated IHC instructions (standard IHC practice). |
| A known high-staining population is completely negative. | A failed staining run or unsuitable section is possible (standard IHC practice); HPA reports high staining for the named populations (HPA tissue IHC). | Check the positive control, reagent sequence and detection system, then repeat with an intact section (standard IHC practice). Avoid calling the tissue HSPE1-negative from one run. |
| Signal is mainly nuclear or along cell borders. | That distribution disagrees with mitochondrial matrix localisation (UniProt P61604) and HPA's granular cytoplasmic profile (HPA tissue IHC). | Recheck compartment assignment against the counterstain and tissue morphology; assess primary-omission and antibody-specific controls (standard IHC practice). |
| Background is diffuse or extracellular. | Nonspecific binding, incomplete washing or endogenous detection activity can produce background (standard IHC practice). | Use a primary-omission control; review blocking, washes and detection chemistry before changing how positive cells are scored (standard IHC practice). |
| Unexpected cells stain strongly. | HPA provides no negative cell list and cautions that the antibodies target protein from more than one gene (HPA tissue IHC). | Identify the cells morphologically and compare controls or an independently validated antibody before assigning the signal to HSPE1 (standard IHC practice). |
| Can the tissue IHC result predict an ICC-IF pattern? | HPA has no ICC-IF images or main subcellular location for HSPE1 in this payload (HPA subcellular); the listed antibodies have no stated ICC status (HPA antibodies). | Use the separate IF/ICC guide and validate localisation in the chosen cells; use mitochondrial matrix localisation as a hypothesis (UniProt P61604). |
Experiment evidence
IHC & IF images showing HSPE1 staining
M06192-1 · IHCIHC analysis of HSP10 using anti-HSP10 antibody (M06192-1). HSP10 was detected in a paraffin-embedded section of human colorectal adenocarci
M06192-1 · IHCIHC analysis of HSP10 using anti-HSP10 antibody (M06192-1). HSP10 was detected in a paraffin-embedded section of human lung adenocarcinoma t
PA1790 · IHCIHC analysis of Cpn10 using anti-Cpn10 antibody (PA1790). Cpn10 was detected in immunocytochemical section of A549 Cell. Enzyme antigen retr
PA1790 · IHCIHC analysis of Cpn10 using anti-Cpn10 antibody (PA1790). Cpn10 was detected in paraffin-embedded section of Human Intestinal Cancer Tissue.
PA1790 · IFIF analysis of Cpn10 using anti-Cpn10 antibody (PA1790). Cpn10 was detected in immunocytochemical section of HepG2 cells. Enzyme antigen ret
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115541-HSPE1/tissue/cerebral+cortex#imid_16390327
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000115541-HSPE1/tissue/adrenal+gland#imid_16390287
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115541-HSPE1/tissue/colon#imid_16390253
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115541-HSPE1/tissue/liver#imid_16390238
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HSPE1 IHC & IF
🧪Run adipose tissue first and require staining in adipocytes (HPA: High in adipocytes). HPA detects HSPE1 in all 45 scored tissues, so there is no validated negative tissue or internal negative cell type; use no-primary and isotype controls for background, and expect nuclei within stained adipocytes to remain unstained (HPA: no negative rows; UniProt P61604: mitochondrial matrix).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: HSPE1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for HSPE1; derive a cell-line control from the positive tissue's cell type (Adipocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only slide, a host- and immunoglobulin class-matched isotype control, and HSPE1-knockout material as a biological negative if available (selected-SKU caption: rabbit primary; standard IHC controls). Quench endogenous peroxidase for chromogenic detection and assess background along the thin adipocyte cytoplasmic rim (selected-SKU caption: HRP/DAB detection; standard adipose IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state a fixative (selected-SKU caption: fixative unreported). The caption uses heat-mediated EDTA retrieval at pH 8.0 on a paraffin section, but does not establish that retrieval is required for adipose tissue (selected-SKU caption: EDTA pH 8.0). The supplied evidence does not establish that frozen sections or IF are easier; in paraffin adipose sections, large lipid spaces leave a thin cytoplasmic rim for scoring (HPA: High in adipocytes; standard paraffin morphology).
HPA tissue IHC evidence for HSPE1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| 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 → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: HSPE1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced HSPE1 IHC Tips
Troubleshoot HSPE1 staining in paraffin sections by checking retrieval, compartmental pattern, controls, and scoring before interpreting differences between samples.
Which retrieval conditions should I try first for HSPE1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M06192-1). That condition precedes 1:50 primary antibody incubation overnight at 4°C in the documented colorectal adenocarcinoma IHC example, so use its staining pattern as a reference for an initial run (datasheet M06192-1). If staining is weak, compare retrieval duration or a second buffer on adjacent sections while keeping antibody incubation and detection constant (standard IHC practice). Judge improvement by a granular cytoplasmic pattern, rather than stronger staining alone, and check that background has not increased (HPA tissue IHC; standard IHC practice).
How should I troubleshoot fixation when HSPE1 staining varies between specimens?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity for this antibody is unknown (datasheet M06192-1). Record each specimen’s fixative, fixation interval, processing history, and section age before comparing staining, because those variables can affect antigen accessibility in IHC (standard IHC practice). Run sections with different processing histories in the same staining batch using EDTA at pH 8.0 and the same detection steps (datasheet M06192-1; standard IHC practice). If a specimen remains weak, use a consistently processed reference section to assess the run, but do not assign the difference to an HSPE1-specific fixation effect without a controlled comparison (standard IHC practice).
What HSPE1 pattern should I expect, and how should I assess diffuse staining?
Expect a granular cytoplasmic IHC pattern, consistent with HSPE1’s mitochondrial matrix location (HPA tissue IHC; UniProt P61604 subcellular location). The precursor has no annotated transmembrane segment, so a crisp plasma-membrane outline is not the expected pattern for this target (UniProt P61604 topology). Compare granularity in cells and regions with documented strong tissue staining, such as adrenal gland glandular cells or adipocytes, while allowing for differences between specimens (HPA tissue IHC). If staining looks uniformly diffuse, inspect adjacent sections for morphology and background, then compare a no-primary control before calling the signal specific (standard IHC practice).
Could processing or modification change which HSPE1 epitope is detected?
UniProt annotates a 102-aa precursor and a chain spanning residues 2–102, with no annotated isoforms (UniProt P61604 processing; UniProt P61604 isoforms). It also lists modified residues, including acetylation at residues 2 and 8 and alternative modifications at residue 40 (UniProt P61604 modified residues). Those annotations do not establish the catalog antibody’s epitope or show that a modification changes its IHC binding (UniProt P61604 modified residues; datasheet M06192-1). When staining differs despite matched processing, compare a second antibody with a documented distinct epitope, if available, and retain a no-primary control to assess background (standard IHC practice).
How can I assess HSPE1 by multiplex IF in a section?
Treat IF as a separate optimisation: the supplied catalog example documents chromogenic IHC in a paraffin section, while fixation for that section is unreported (datasheet M06192-1). For a defined cell population, pair HSPE1 with an appropriate cell-type marker; glandular cells in adrenal gland are one documented high-staining population to examine (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and assess tissue autofluorescence in each channel, favouring a longer-wavelength HSPE1 channel when shorter-wavelength background is prominent (standard IF practice). Because HSPE1 is in the mitochondrial matrix, optimise permeabilisation to give antibody access across cellular and mitochondrial membranes, then check whether punctate signal aligns with a mitochondrial marker (UniProt P61604 subcellular location; standard IF practice).
What should I change when DAB or cytoplasmic background obscures HSPE1?
First compare the stained section with a no-primary control and inspect whether unwanted colour follows tissue edges, damaged areas, or the expected granular cytoplasmic pattern (standard IHC practice; HPA tissue IHC). The documented example used 10% goat serum blocking, 1:50 primary antibody overnight at 4°C, and an HRP/DAB detection system (datasheet M06192-1). Include an endogenous peroxidase block as a general HRP-IHC step, and assess whether changing wash stringency or primary incubation reduces nonspecific signal (standard IHC practice). Keep retrieval at EDTA pH 8.0 during this comparison so that a background change is easier to attribute to blocking or detection (datasheet M06192-1; standard IHC practice).
How should I score HSPE1 across samples with different cellular makeup? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue area before scoring, because HSPE1 staining can differ between cell types within a section (HPA tissue IHC; standard IHC practice). Record the percentage of positive cells and intensity, or calculate an H-score from intensity categories, using identical thresholds and exposure or imaging settings across samples (standard IHC practice). If counting positive cells, report density per mm² of viable, annotated tissue; for an H-score, report the fraction of the specified cell population assessed (standard IHC practice). Normalise comparisons to the number of eligible cells or their viable area, and record granular cytoplasmic signal separately from diffuse background (HPA tissue IHC; standard IHC practice).
When is a strong HSPE1 stain likely to be an artefact?
A credible HSPE1 signal should be granular and cytoplasmic, consistent with mitochondrial matrix localisation, rather than a sharp membrane or exclusively nuclear pattern (HPA tissue IHC; UniProt P61604 subcellular location). Check whether the stained cells fit the tissue context; HPA reports high staining in adrenal gland glandular cells and adipocytes, but those examples do not establish every specimen’s expected level (HPA tissue IHC). Edge accentuation, necrotic regions, and colour persisting in a no-primary control warrant assessment for section or endogenous-enzyme artefact (standard IHC practice). Interpret apparent positives cautiously because HPA rates its tissue IHC evidence Supported yet cautions that its antibody targets protein from more than one gene (HPA tissue IHC).
Boster reagents
Best HSPE1 / 10 kDa heat shock protein, mitochondrial IHC Antibodies
Both antibodies list human, mouse, and rat reactivity (catalog). Their image captions document paraffin-section IHC in all three species and IF in human samples (IHC and IF image captions).
Real IHC data
Anti-HSP10 Rabbit Monoclonal Antibody
Cat # M06192-1
Real IHC data
Anti-Cpn10/HSPE1 Antibody ®
Cat # PA1790
M06192-1 has paraffin-section IHC images from human colorectal, lung, prostate, and liver cancers (M06192-1 IHC image captions). PA1790 has paraffin-section IHC images from human intestinal cancer and mouse and rat intestine, plus IF images from human cells and intestinal cancer tissue (PA1790 IHC and IF image captions).
Which to pick: For human tissue IHC, choose the rabbit monoclonal M06192-1 if its documented EDTA pH 8.0 retrieval and 1:50 dilution suit your workflow (catalog; M06192-1 IHC image captions). For IF/ICC, PA1790 has documented human cell IF images; for tissue IHC across species, it has documented human, mouse, and rat paraffin-section images using citrate pH 6 retrieval and 1 μg/ml antibody (PA1790 IF and IHC image captions). The paraffin-section captions do not report the fixative for either SKU (M06192-1 and PA1790 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P61604 (CH10_HUMAN, 10 kDa heat shock protein, mitochondrial).
- Human Protein Atlas. HSPE1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. HSPE1 subcellular location (ICC-IF): Variation in transcript expression is correlated to the cell cycle..
- Human Protein Atlas. HSPE1 antibody validation summary (3 antibodies).
- Systematic verification of bladder cancer-associated tissue protein biomarker candidates in clinical urine specimens. Oncotarget 2018 — PMC6089400.
- Comparative proteomics analysis suggests that placental mitochondria are involved in the development of pre-eclampsia. PloS one 2013 — PMC3650054.
- Quantitative proteomic analysis reveals potential diagnostic markers and pathways involved in pathogenesis of renal cell carcinoma. Oncotarget 2014 — PMC3964225.
- Transcriptomic and Proteomic Analysis of Clear Cell Foci (CCF) in the Human Non-Cirrhotic Liver Identifies Several Differentially Expressed Genes and Proteins with Functions in Cancer Cell Biology and Glycogen Metabolism. Molecules (Basel, Switzerland) 2020 — PMC7570661.
- PubMed PMID:7914093 — UniProt-cited evidence.
- PubMed PMID:7916212 — UniProt-cited evidence.
- PubMed PMID:12483302 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HSPE1 (10 kDa heat shock protein, mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrial matrix (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M06192-1). Validated IHC antibody: M06192-1 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M06192-1); 2. Fixation: Image fixative and duration unreported (datasheet M06192-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet M06192-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M06192-1); 8. Primary antibody: Rabbit monoclonal (clone 19H75) anti-HSPE1, 1:50 (datasheet M06192-1); 9. Primary incubation: Overnight at 4 °C (datasheet M06192-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M06192-1); Caveat: HPA staining may reflect proteins from multiple genes (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans 2–102 (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt P61604, HPA tissue IHC · Supported, HPA cell-line RNA · Variation in transcript expression is correlated to the cell cycle., Datasheet M06192-1, PMC6089400, PMC3650054. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HSPE1/immunohistochemistry-ihc-guide.