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GRPEL1 / GrpE protein homolog 1, mitochondrial · IHC design guide
Design Immunohistochemistry for GRPEL1
Plan chromogenic paraffin IHC for GRPEL1 using the catalog antibody at 2–5 μg/ml (datasheet A12830-1). Appendix glandular cells provide a high-staining reference (HPA tissue IHC); granular cytoplasmic staining is consistent with its mitochondrial matrix location (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GRPEL1 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 cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Glandular cells show granular cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A12830-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Hippocampus+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding may confound staining (HPA tissue IHC) | |
| Regulation | Low tissue specificity; regulation unreported (HPA tissue IHC; UniProt) | |
| Isoform / epitope | No isoforms; mature chain 28–217; no extracellular domain (UniProt) |
Section 1
Recommended GRPEL1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by two published chromogenic GRPEL1 IHC protocols using human PDAC tissue microarrays (PMC13392447; PMC12894993).
| Sample | Paraffin-embedded human lymphadenoma tissue; fixative not specified (datasheet A12830-1) |
| Fixation | Image fixative and duration unreported (datasheet A12830-1); 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 A12830-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A12830-1) |
| Primary antibody | Rabbit anti-GRPEL1, 2-5 μg/ml (datasheet A12830-1) |
| Primary incubation | Overnight at 4 °C (datasheet A12830-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A12830-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GRPEL1-positive staining in glandular cells of adrenal gland (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 pH 8.0 retrieval (datasheet: A12830-1); a published PDAC protocol used boiling EDTA pH 8.0 for 15–20 min (PMC13392447).
Section 2
What Is the Expected GRPEL1 Staining Pattern?
GRPEL1 is a mitochondrial matrix protein with no transmembrane segment (UniProt Q9HAV7 localization and topology). In paraffin IHC, expect granular cytoplasmic staining in glandular cells, including those of the adrenal gland, appendix and colon (HPA tissue IHC: High; general cytoplasmic granular pattern). HPA rates the tissue pattern Supported, reports medium agreement with RNA data, and notes presumed off target staining that was disregarded (HPA tissue IHC reliability).
| Granular cytoplasmic stain in adrenal, appendix or colon glandular cells. | This fits the reported IHC pattern and High staining in those cell populations (HPA tissue IHC). Mitochondrial matrix localization supports a punctate or granular cytoplasmic interpretation, although chromogenic IHC alone does not prove that each granule is a mitochondrion (UniProt Q9HAV7 localization; general IHC practice). |
| Predominantly nuclear, cell surface or extracellular staining, with little granular cytoplasm. | Reassess specificity and slide quality: these compartments do not match the main mitochondrial location (UniProt Q9HAV7; HPA subcellular ICC-IF). HPA also reports nucleoplasmic signal as uncertain, so a small nuclear component is inconclusive; strong nuclear-only IHC should not be called the expected pattern (HPA subcellular ICC-IF). |
| Strong staining of hippocampal glia or skeletal myocytes. | These cell populations were Not detected in the supplied tissue IHC observations (HPA tissue IHC). Check for cross-reactivity or detection activity before assigning the stain to GRPEL1; HPA reports that presumed off target staining was disregarded in its tissue assessment (HPA tissue IHC reliability). |
| Diffuse color across cells, stroma and blank areas, obscuring granules. | The distribution does not resemble HPA's general granular cytoplasmic pattern (HPA tissue IHC). Uneven washing, excess detection reagent or endogenous enzyme activity can raise chromogenic background (general IHC practice). A no-primary control helps distinguish detection background from primary antibody-associated staining (general IHC practice). |
| No convincing stain in glandular cells of an expected positive tissue. | A blank result conflicts with reported High staining in adrenal, appendix or colon glandular cells (HPA tissue IHC). Review tissue preservation, retrieval and detection controls before interpreting absence as biology (general IHC practice). HPA's Supported rating is evidence for a pattern, not a guarantee of signal in every section (HPA tissue IHC reliability). |
💡Expected GRPEL1 appearanceCall a positive result when glandular cells show distinct granular cytoplasmic staining, potentially strong in HPA High tissues; widespread diffuse or dominant surface stain is suspect (HPA tissue IHC; UniProt Q9HAV7 localization).
| Compartment and topology | GRPEL1 is assigned to the mitochondrial matrix and has no transmembrane segment (UniProt Q9HAV7). Score cytoplasmic granules in the appropriate cells; surface outlining alone is discordant with that assignment (UniProt Q9HAV7; HPA tissue IHC pattern). |
| Protein processing | UniProt annotates a mature chain spanning residues 28–217 and a transit peptide keyword (UniProt Q9HAV7 processing and keywords). This supports mitochondrial targeting; the supplied record does not identify the IHC antibody's epitope, so processing cannot predict its staining strength. |
| Tissue distribution | HPA calls RNA tissue specificity Low and describes general cytoplasmic protein expression, while reporting both High and Not detected cell populations (HPA tissue IHC). Judge signal by the named cell population and compartment, rather than requiring equal intensity across all tissues. |
| Antibody validation | HPA036647 is listed as IHC Supported, and the overall tissue profile is Supported with medium agreement between staining and RNA data (HPA antibodies; HPA tissue IHC reliability). Treat an unexpected distribution as a finding to verify, especially given HPA's disregarded presumed off target staining (HPA tissue IHC reliability). |
| IF/ICC Q&A: What should fluorescence show? | Mainly mitochondrial signal is supported; additional nucleoplasmic localization is uncertain (HPA subcellular ICC-IF). Use the separate IF/ICC guide for its experimental workflow. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive glandular cells are blank. | The slide may have weak overall detection; High staining is reported for adrenal, appendix and colon glandular cells (HPA tissue IHC). | Check a positive control on the same run, then review retrieval, primary antibody conditions and detection performance (general IHC practice). Avoid calling the tissue negative until the run controls work. |
| Every cell compartment has a uniform brown haze. | Diffuse background obscures the reported granular cytoplasmic pattern (HPA tissue IHC). Excess reagent or incomplete washing can contribute (general IHC practice). | Compare with a no-primary control; review blocking, reagent concentration, wash steps and exposure to chromogen (general IHC practice). Re-score only where cell borders and granules remain interpretable. |
| Strong signal concentrates at nuclei or cell surfaces. | This conflicts with the main mitochondrial assignment; nucleoplasmic signal is reported only as uncertain (UniProt Q9HAV7; HPA subcellular ICC-IF). | Compare the same section with expected positive cell populations and appropriate controls; assess whether granular cytoplasm is also present (HPA tissue IHC; general IHC practice). Do not score an isolated nuclear or surface pattern as confirmed GRPEL1. |
| Hippocampal glia or skeletal myocytes stain strongly. | Both populations were Not detected in HPA's tissue IHC observations; presumed off target staining was also reported and disregarded (HPA tissue IHC). | Recheck cell identity and controls, then seek independent specificity evidence before interpreting the stain as GRPEL1 (general IHC practice). Treat HPA's Not detected calls as observations, not universal biological negatives. |
| Color appears in a no-primary control. | Signal without primary antibody indicates background from the detection system or endogenous tissue activity (general IHC practice). | Review the detection chemistry and its blocking step, then repeat the control alongside the test section (general IHC practice). Only primary-dependent granular staining should contribute to the GRPEL1 call. |
| Granules are present, but staining is weaker than another tissue. | HPA reports different cell-level staining categories, including High and Low, despite Low RNA tissue specificity (HPA tissue IHC). | Identify the scored cell population and compare it with that tissue's HPA observation; confirm run controls before changing conditions (HPA tissue IHC; general IHC practice). Do not impose one intensity cutoff across tissues. |
Experiment evidence
IHC & IF images showing GRPEL1 staining
A12830-1 · IHCIHC analysis of GRPEL1 using anti-GRPEL1 antibody (A12830-1). GRPEL1 was detected in a paraffin-embedded section of human lymphadenoma tissu
A12830-1 · IHCIHC analysis of GRPEL1 using anti-GRPEL1 antibody (A12830-1). GRPEL1 was detected in a paraffin-embedded section of human appendiceal adenoc
A12830-1 · IFIF analysis of GRPEL1 using anti-GRPEL1 antibody (A12830-1). GRPEL1 was detected in an immunocytochemical section of PC-3 cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109519-GRPEL1/tissue/cerebral+cortex#imid_9124684
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109519-GRPEL1/tissue/colon#imid_9124610
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109519-GRPEL1/tissue/liver#imid_9124595
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109519-GRPEL1/tissue/kidney#imid_9124604
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036647; green = GRPEL1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036647; green = GRPEL1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GRPEL1 IHC & IF
🧪Run adrenal gland first and require strong staining in its glandular cells (HPA: High in adrenal gland glandular cells); use hippocampus glial cells as the negative tissue (HPA: Not detected in hippocampus glial cells). On the adrenal slide, treat surrounding nonglandular cells as internal background comparators, recording their staining rather than assuming they are target-negative (HPA: High is specified for glandular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Hippocampus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GRPEL1 in A-431, U-251MG, U2OS, Sperm, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control for the rabbit primary; and a GRPEL1 knockout or validated peptide-block control (selected-SKU tissue-IHC caption: rabbit anti-GRPEL1 primary). Check adrenal sections for endogenous peroxidase and biotin signal before interpreting brown staining from the biotin-based DAB detection system (selected-SKU tissue-IHC caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption: fixative not stated). The reported paraffin-section method uses heat-mediated EDTA retrieval at pH 8.0, so validate retrieval with that condition before scoring; the supplied evidence does not establish whether frozen sections or IF are easier (selected-SKU tissue-IHC caption: EDTA retrieval; HPA subcellular: ICC-IF images). In adrenal gland, assess endogenous peroxidase and biotin background as potential artefacts of the reported DAB/SABC method (selected-SKU tissue-IHC caption: SABC and DAB).
HPA tissue IHC evidence for GRPEL1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cilia axoneme) | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced GRPEL1 IHC Tips
Use compartment, tissue context and matched controls to troubleshoot GRPEL1 staining in paraffin sections; interpret the published patterns with their stated reliability.
What retrieval should I use when GRPEL1 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA buffer at pH 8.0 (datasheet A12830-1). The catalog antibody detected GRPEL1 in a paraffin section after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (caption A12830-1). If staining remains weak, check heating consistency, section adhesion and the positive control before changing the retrieval buffer (standard IHC practice). Compare any fallback retrieval condition on adjacent sections, keeping detection and exposure to DAB consistent (standard IHC practice). Judge improvement by a granular cytoplasmic pattern rather than diffuse staining alone (HPA tissue IHC profile; UniProt Q9HAV7 subcellular location).
Could fixation explain weak or uneven GRPEL1 staining?
Target-specific fixation sensitivity is unknown because the selected tissue caption reports a paraffin section without stating its fixative (caption A12830-1). Record the fixative and processing history for each section, and compare samples processed together when troubleshooting uneven signal (standard IHC practice). Keep the established EDTA pH 8.0 retrieval and 2 μg/ml primary condition constant while assessing processing differences (datasheet A12830-1; caption A12830-1). Examine morphology and staining across the section, since folds, drying and uneven processing can complicate interpretation (standard IHC practice). Do not assign a GRPEL1-specific fixation effect from its mitochondrial location or modified residues (UniProt Q9HAV7).
Which staining compartment is plausible for GRPEL1 in tissue sections?
Expect a predominantly granular cytoplasmic pattern in chromogenic tissue sections (HPA tissue IHC profile). GRPEL1 is a mitochondrial matrix protein without an annotated transmembrane segment, which supports a mitochondrial interpretation of fine cytoplasmic granules (UniProt Q9HAV7 subcellular location and topology). Compare the pattern with tissue morphology and a matched negative control before calling coarse pigment or diffuse DAB deposition positive (standard IHC practice). HPA also reports a nucleoplasmic location in cell imaging, but marks that additional location uncertain (HPA subcellular summary). Treat nuclear-only tissue staining cautiously and seek independent support before scoring it as GRPEL1 (HPA subcellular summary; HPA tissue IHC reliability).
How should epitope uncertainty affect GRPEL1 IHC interpretation?
The record lists 0 isoforms and a processed chain spanning residues 28–217 (UniProt Q9HAV7 processing and isoforms). It also lists modified lysines, including alternate modifications at residue 94, but supplies no antibody epitope mapping (UniProt Q9HAV7 modified residues; caption A12830-1). Therefore, do not attribute altered staining to a particular modification or mature-chain epitope without direct antibody characterization (standard IHC interpretation). When comparing samples, hold retrieval, antibody concentration and detection constant, then inspect whether signal retains the expected granular cytoplasmic distribution (datasheet A12830-1; HPA tissue IHC profile). An independent antibody with a known, distinct epitope can help test specificity if its validation is available (standard IHC practice).
How can IF help assess an ambiguous GRPEL1 IHC pattern?
Use IF as a separate corroborating experiment when chromogenic staining lacks clear intracellular detail (standard IHC/IF practice). Multiplex GRPEL1 with a marker identifying the expected glandular cell population, since several glandular cell populations show high tissue staining in the HPA profile (HPA tissue IHC profile). Choose a fluorophore channel away from the tissue’s strongest autofluorescence and include single-channel and no-primary controls (standard IF practice). Because GRPEL1 is in the mitochondrial matrix, assess whether fixation and permeabilisation allow antibody access across mitochondrial membranes while preserving punctate morphology (UniProt Q9HAV7 subcellular location; standard IF practice). Nuclear IF signal warrants caution because HPA labels nucleoplasmic localisation uncertain (HPA subcellular summary).
What controls distinguish GRPEL1 signal from chromogenic background?
Run a no-primary control and inspect the same tissue compartments for nonspecific secondary or detection-system staining (standard IHC practice). The selected caption used 10% goat serum blocking, a biotinylated secondary and a streptavidin–biotin complex with DAB (caption A12830-1). Include an appropriate endogenous biotin control when using that detection chemistry, and block endogenous peroxidase as a general DAB workflow step (standard IHC practice). Titrate the primary around the documented 2 μg/ml condition while keeping retrieval and development comparable (caption A12830-1; standard IHC practice). Distrust uniform haze or staining confined to damaged regions when the expected tissue pattern is granular cytoplasmic (HPA tissue IHC profile; standard IHC interpretation).
How should I quantify GRPEL1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; the tissue reference describes granular cytoplasmic GRPEL1 staining (HPA tissue IHC profile). For chromogenic sections, record the percentage of positive viable cells and staining intensity, then calculate an H-score from intensity categories if that scale suits the study (standard IHC scoring practice). Alternatively, quantify positive-cell density per mm² of viable tissue when cell counts and sampled area can be measured reliably (standard IHC image analysis practice). Normalise comparisons to the same cell type, sampled area, section processing and DAB development conditions (standard IHC practice). Report background thresholds and exclude folds or necrosis consistently across groups (standard IHC image analysis practice).
When is an apparent GRPEL1-positive cell likely to be artefactual?
Give greatest weight to granular cytoplasmic staining in morphologically intact cells, consistent with the tissue profile and mitochondrial matrix localisation (HPA tissue IHC profile; UniProt Q9HAV7 subcellular location). Check cell identity against the sampled tissue: HPA reports high staining in several glandular cell populations, while hippocampal glial cells and skeletal muscle myocytes were not detected in its listed examples (HPA tissue IHC profile). Treat isolated nuclear-only signal, section-edge accentuation, necrotic areas or pigment-like deposits as findings requiring controls (HPA subcellular summary; standard IHC interpretation). Compare no-primary and peroxidase controls when DAB staining could reflect detection background (standard IHC practice). HPA rates tissue evidence Supported while noting presumed off-target binding, so corroborate unexpected patterns independently (HPA tissue IHC reliability).
Boster reagents
Best GRPEL1 / GrpE protein homolog 1, mitochondrial IHC Antibodies
A12830-1 has IHC data from human paraffin tissue and IF/ICC data from PC-3 cells; its listed reactivity is human (catalog reactivity; IHC and IF image captions).
Real IHC data
Anti-GRPEL1 Antibody ®
Cat # A12830-1
A12830-1 is listed for human IHC, with images of paraffin sections from lymphadenoma, appendiceal adenocarcinoma, colonic adenocarcinoma, and esophageal squamous carcinoma (catalog applications/reactivity; IHC image captions). The same SKU is listed for IF/ICC and shown staining PC-3 cells (catalog applications; IF image caption).
Which to pick: Choose A12830-1 for tissue IHC: its own image caption shows staining in a human paraffin section after EDTA pH 8 retrieval, using 2 μg/ml primary antibody (A12830-1 IHC image caption). For IF/ICC, the same SKU is listed at 5 μg/ml and shown in PC-3 cells (catalog applications/dilution; A12830-1 IF image caption). No cross-species choice is supported: A12830-1 lists human reactivity only; clonality and the paraffin tissue fixative are unreported (catalog reactivity/clone; A12830-1 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9HAV7 (GRPE1_HUMAN, GrpE protein homolog 1, mitochondrial).
- Human Protein Atlas. GRPEL1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. GRPEL1 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoplasm..
- Human Protein Atlas. GRPEL1 antibody validation summary (1 antibodies).
- Inference of upstream-mutation and metabolomic-signature causality identifies prognostic biomarkers and therapeutic targets in pancreatic cancer. Nature communications 2026 — PMC13392447.
- The LIV-1-GRPEL1 axis adjusts cell fate during anti-mitotic agent-damaged mitosis. EBioMedicine 2019 — PMC6945280.
- c-Myc/GRPEL1 maintains fatty acid synthesis via FASN to support PDAC cell proliferation. Cell death & disease 2026 — PMC12894993.
- Inter-organellar and systemic responses to impaired mitochondrial matrix protein import in skeletal muscle. Communications biology 2022 — PMC9534917.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:11311562 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GRPEL1 (GrpE protein homolog 1, mitochondrial). Expected localisation Granular cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A12830-1). Validated IHC antibody: A12830-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Hippocampus. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lymphadenoma tissue; fixative not specified (datasheet A12830-1); 2. Fixation: Image fixative and duration unreported (datasheet A12830-1); 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 A12830-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A12830-1); 8. Primary antibody: Rabbit anti-GRPEL1, 2-5 μg/ml (datasheet A12830-1); 9. Primary incubation: Overnight at 4 °C (datasheet A12830-1). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target binding may confound staining (HPA tissue IHC); Isoform / epitope: No isoforms; mature chain 28–217 ; no extracellular domain (UniProt); Regulation: Low tissue specificity; regulation unreported (HPA tissue IHC; UniProt). Evidence: UniProt Q9HAV7, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A12830-1, PMC13392447, PMC12894993. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GRPEL1/immunohistochemistry-ihc-guide.