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PGAM1 / Phosphoglycerate mutase 1 · IHC design guide
Design Immunohistochemistry for PGAM1
Plan PGAM1 chromogenic IHC in paraffin sections using cytoplasmic staining in most tissues as the expected pattern (HPA tissue IHC). Use kidney tubules as a high-staining reference (HPA tissue IHC), start the catalog antibody at 2–5 μg/ml (datasheet: A04470-2), and assess possible cross-gene staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PGAM1 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 | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal in most tissues; high in kidney tubules (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04470-2) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
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 A04470-2) | |
| Caveat | Possible cross-gene antibody staining (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 0 isoforms annotated; chain spans residues 2–254 (UniProt) |
Section 1
Recommended PGAM1 IHC & IF Protocols
The catalog antibody protocol uses heat retrieval in EDTA pH 8.0 (datasheet A04470-2). Four published PGAM1 IHC protocols provide sample specific comparisons (PMC5958715; PMC2873438; PMC5858104; PMC5559954).
| Sample | Paraffin-embedded human testis cancer tissue; fixative not specified (datasheet A04470-2) |
| Fixation | Image fixative and duration unreported (datasheet A04470-2); 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 A04470-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04470-2) |
| Primary antibody | Rabbit anti-PGAM1, 2-5 μg/ml (datasheet A04470-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04470-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04470-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PGAM1-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A04470-2). Published protocols using other antibodies describe citrate retrieval (PMC5958715; PMC2873438; PMC5858104; PMC5559954).
Section 2
What Is the Expected PGAM1 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic PGAM1 staining, with strong staining in kidney tubule cells and moderate staining in selected glandular, respiratory epithelial, and neural compartments (HPA tissue IHC). PGAM1 has no transmembrane segment (UniProt P18669 topology). Treat this pattern as a guide: HPA rates the tissue IHC profile Approved, with medium consistency against RNA data, pending external verification, and a warning that the antibody may target proteins from more than one gene (HPA tissue IHC).
| Strong cytoplasmic staining in kidney tubule cells, with tissue structure still visible. | This matches the clearest listed positive reference: kidney tubule cells stain High (HPA tissue IHC). Judge staining within tubule cells, rather than treating color anywhere in the section as a positive result. The HPA profile carries a multi-gene targeting caution (HPA tissue IHC). |
| Moderate cytoplasmic staining in adrenal glandular cells, bronchial respiratory epithelial cells, or cerebellar granular-layer cells. | These cell populations are listed at Medium intensity (HPA tissue IHC). A lighter result than kidney tubules can therefore fit the reported tissue pattern. Compare the stained cell population and compartment as well as intensity; HPA describes cytoplasmic expression in most tissues (HPA tissue IHC). |
| A nuclear-only or sharply membrane-restricted IHC pattern, without the expected cytoplasmic staining. | This departs from the reported tissue IHC profile and warrants a compartment check before scoring the slide positive (HPA tissue IHC; UniProt P18669 topology). Nuclear signal alone is not proof of an artefact across applications: HPA reports approved nucleoplasmic localization in ICC-IF, with a multi-gene antibody caution (HPA subcellular ICC-IF). |
| Strong color in skeletal muscle myocytes or adipocytes, or similar color across unrelated cell types. | Those cell types are listed as Not detected in the sampled HPA tissue IHC views (HPA tissue IHC). Check for cross-reactivity or nonspecific chromogen deposition before assigning PGAM1 positivity. Endogenous detection activity is another general chromogenic-IHC possibility; the slide pattern alone cannot identify which cause applies. |
| Hazy, diffuse color across tissue and empty areas, or no staining in kidney tubule cells. | Diffuse color that ignores cell boundaries is unsuitable for compartment scoring. An unstained known-positive population conflicts with the High kidney-tubule reference (HPA tissue IHC). Review assay controls and staining conditions before interpreting other tissues as negative; one failed positive control cannot establish their PGAM1 status. |
💡Expected PGAM1 appearanceCall an IHC result convincing when kidney tubule cells show High, predominantly cytoplasmic staining (HPA tissue IHC); color confined to an unexpected compartment or prominent in HPA-listed Not detected cell types needs investigation, especially given HPA’s multi-gene targeting caution (HPA tissue IHC).
| Tissue and cell type | Intensity is cell-specific: kidney tubule cells are High; several glandular, respiratory epithelial, and neural populations are Medium; skeletal muscle myocytes and adipocytes are Not detected (HPA tissue IHC). Use the sampled cell population when choosing comparison fields. |
| IHC evidence strength | The tissue profile is Approved but has medium staining-to-RNA consistency, is pending external verification, and warns of proteins from more than one gene being targeted (HPA tissue IHC). Treat an unexpected positive as provisional until independently checked. |
| Protein topology | PGAM1 has no annotated transmembrane segment or signal peptide (UniProt P18669 topology and processing). This is compatible with the reported cytoplasmic tissue IHC profile (HPA tissue IHC); topology alone does not validate an individual stained structure. |
| IF/ICC question: should nuclear fluorescence be expected? | HPA reports mainly nucleoplasmic localization and additional mid-piece localization in ICC-IF, both marked approved, while cautioning that antibodies may target proteins from multiple genes (HPA subcellular ICC-IF). Interpret that IF observation within its own application; the tissue IHC profile is cytoplasmic (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney tubule cells are unstained. | The positive reference has failed: these cells are High in HPA tissue IHC. The image alone does not identify the failed assay step. | Check a known-positive section, reagent delivery, retrieval conditions, antibody dilution, and detection controls using the IHC-validated antibody’s documented procedure. Restore the positive-control signal before scoring study sections as negative. |
| Color is widespread, including cell types listed as Not detected. | Nonspecific binding, cross-reactivity, or endogenous chromogenic detection activity are possible general IHC causes. HPA also warns that its staining antibody may target proteins from more than one gene (HPA tissue IHC). | Compare the no-primary and detection controls, review blocking and washing, and assess whether staining follows cells rather than tissue edges or deposits. Confirm unexpected cell-type positives independently before reporting PGAM1 expression. |
| A slide is predominantly nuclear or membrane-stained. | That pattern does not match HPA’s cytoplasmic tissue IHC profile (HPA tissue IHC). Approved nucleoplasmic ICC-IF staining exists, so compartment findings depend on the application (HPA subcellular ICC-IF). | Inspect matched controls and morphology, then compare cytoplasmic staining in kidney tubule cells (HPA tissue IHC). If nuclear-only IHC persists, describe the observed pattern without assigning it to PGAM1 on compartment alone. |
| Only weak, hazy color is visible throughout the section. | Diffuse background can obscure cellular staining in chromogenic IHC; its presence does not establish PGAM1 localization. HPA’s reference pattern is cytoplasmic in most tissues (HPA tissue IHC). | Review no-primary control, wash and blocking steps, chromogen development, and counterstain. Score only signal with recognizable cell boundaries; reassess the section after background is controlled. |
| Skeletal muscle myocytes stain strongly. | HPA lists skeletal muscle myocytes as Not detected, and UniProt reports PGAM1 as not found in muscle (HPA tissue IHC; UniProt P18669 tissue specificity). Strong staining there is an unexpected result. | Check cell identity, background controls, and antibody specificity before calling the myocytes PGAM1-positive. Compare the same run with kidney tubules as the HPA High positive reference (HPA tissue IHC). |
| IF/ICC shows nucleoplasmic signal while tissue IHC appears cytoplasmic. | HPA reports approved nucleoplasmic ICC-IF localization and a cytoplasmic tissue IHC profile; both carry an antibody multi-gene caution (HPA subcellular ICC-IF; HPA tissue IHC). | Record the application and compartment with each image. Use the tissue IHC pattern to interpret paraffin sections, and assess IF/ICC in its separate guide; do not force the two images into one compartment call. |
Experiment evidence
IHC & IF images showing PGAM1 staining
A04470-2 · IHCIHC analysis of PGAM1 using anti-PGAM1 antibody (A04470-2). PGAM1 was detected in a paraffin-embedded section of human testis cancer tissue.
A04470-2 · IHCIHC analysis of PGAM1 using anti-PGAM1 antibody (A04470-2). PGAM1 was detected in a paraffin-embedded section of human thyroid cancer tissue
A04470-2 · IFIF analysis of PGAM1 using anti-PGAM1 antibody (A04470-2). PGAM1 was detected in an immunocytochemical section of Hela cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000171314-PGAM1/tissue/cerebral+cortex#imid_14462772
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000171314-PGAM1/tissue/colon#imid_14462682
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000171314-PGAM1/tissue/liver#imid_14462691
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000171314-PGAM1/tissue/kidney#imid_14462700
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA042528; green = PGAM1, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA042528; green = PGAM1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PGAM1 IHC & IF
🧪Run kidney first: cells in tubules should stain strongly (HPA: High in kidney tubule cells). Use skeletal muscle myocytes as the negative tissue (HPA: Not detected in skeletal muscle myocytes); on the kidney slide, assess adjacent non-tubular areas for background without assuming they are PGAM1-negative.
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PGAM1 in CACO-2, PC-3, Hep-G2, U2OS, ASC52telo, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control for the rabbit primary (selected-SKU caption: rabbit anti-PGAM1); and, if available, PGAM1-knockout material or immunizing-peptide competition (standard IHC controls). Quench endogenous peroxidase and check for residual DAB background in kidney tubules (standard chromogenic IHC practice).
⚠️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 caption: fixative unreported). The demonstrated paraffin-section procedure used heat retrieval in EDTA at pH 8.0, but the evidence does not establish whether retrieval is essential (selected-SKU caption: EDTA retrieval). The supplied evidence does not show that frozen sections or IF are easier; for IF/ICC, HPA reports approved nucleoplasmic localization while cautioning that its result is based on antibodies targeting multiple genes (HPA: subcellular summary).
HPA tissue IHC evidence for PGAM1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification. 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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PGAM1 IHC Tips
Troubleshoot PGAM1 staining in paraffin sections by checking retrieval, compartment, controls, and scoring before interpreting differences between specimens.
What retrieval should I try first when PGAM1 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for this paraffin-section IHC assay (datasheet A04470-2). The selected tissue image used that retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody, so preserve those conditions while checking whether retrieval is the source of weak staining (datasheet A04470-2). Include kidney tubules as a positive tissue reference because their PGAM1 staining is reported as high (HPA: High in kidney tubule cells). If staining remains weak, compare a second retrieval condition on serial sections, keeping detection and imaging settings matched (standard IHC practice).
Could fixation explain inconsistent PGAM1 staining between paraffin sections?
The selected paraffin-section caption does not state a fixative, so target-specific sensitivity to fixation is unknown (datasheet A04470-2: fixative not stated). Record the fixative, time to fixation, duration, and processing history for each specimen before attributing differences to PGAM1 abundance (standard IHC practice). Run matched sections through the same EDTA pH 8.0 retrieval and staining sequence used for the catalog antibody (datasheet A04470-2). If staining still differs, compare a consistently processed positive control in the same run and treat specimen-specific fixation effects as a possibility, not an established PGAM1 behavior (standard IHC practice).
How should I interpret cytoplasmic staining alongside reported nuclear PGAM1 signal?
For tissue IHC, use cytoplasmic staining as the reference pattern because HPA describes cytoplasmic expression in most tissues (HPA: tissue IHC profile). HPA also reports mainly nucleoplasmic signal in ICC/IF, but warns that its subcellular antibodies target proteins from multiple genes (HPA: subcellular summary). Assess nuclear and cytoplasmic DAB separately against the counterstain and against omission controls before calling a compartment positive (standard IHC practice). PGAM1 has no annotated transmembrane segment, while UniProt supplies no subcellular annotation; neither fact establishes that nuclear DAB in a tissue section is PGAM1-specific (UniProt P18669 topology; UniProt P18669 subcellular record).
Can an unusual staining pattern indicate a PGAM1 isoform or altered epitope?
UniProt lists 0 isoforms for PGAM1, so an unexpected IHC pattern cannot be assigned to a documented PGAM1 splice isoform from this record (UniProt P18669 isoforms). The record lists modified residues, including phosphoserines at 14 and 23, but provides no antibody epitope map (UniProt P18669 modified residues; datasheet A04470-2: epitope not stated). Compare serial sections with the same retrieval and primary concentration before treating a staining shift as biological (standard IHC practice). Where specificity matters, use an independently validated reagent or an appropriate loss-of-target control; do not infer modification-sensitive binding without direct epitope evidence (standard IHC practice).
How can IF help resolve ambiguous PGAM1 localisation seen by chromogenic IHC?
Use IF as a separate localisation check and multiplex PGAM1 with a marker that identifies the cell population being evaluated in IHC, such as kidney tubule cells (HPA: High in kidney tubule cells). Select a fluorophore channel with low tissue autofluorescence and include single-stain and no-primary controls before comparing compartments (standard IF practice). PGAM1 has no annotated transmembrane segment, so assess intracellular epitopes with a permeabilisation step and verify that it preserves the morphology needed to distinguish cytoplasm from nuclei (UniProt P18669 topology; standard IF practice). Interpret nucleoplasmic IF cautiously because HPA's subcellular result carries a multiple-gene antibody warning (HPA: subcellular summary).
What should I check when PGAM1 DAB staining appears diffuse or patchy?
First inspect the no-primary section for detection-system background, and block endogenous peroxidase before applying the HRP and DAB workflow (standard chromogenic IHC practice). The selected PGAM1 image used 10% goat serum blocking, a peroxidase-conjugated secondary, and DAB development; those details are assay evidence, not proof that every brown deposit is PGAM1 (datasheet A04470-2). Titrate the primary around its reported 2 μg/ml condition while holding retrieval and development constant (datasheet A04470-2; standard IHC practice). Compare suspect staining with tissue morphology, section edges, and the no-primary control before scoring it (standard IHC practice).
How should I score PGAM1 across sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring; HPA's tissue IHC profile describes predominantly cytoplasmic expression and high staining in kidney tubule cells (HPA: tissue IHC profile; HPA: High in kidney tubule cells). For intensity-based comparisons, calculate a cytoplasmic H-score as 1 × % weak + 2 × % moderate + 3 × % strong within the predefined viable population (standard IHC practice). Report the percentage of positive cells separately, or positive-cell density per mm² when counts are needed (standard IHC practice). Normalize to the number or area of eligible viable cells, and keep exposure, DAB development, and thresholds consistent across sections (standard IHC practice).
When is a PGAM1-positive cell convincing rather than an artefact?
A convincing tissue-IHC result has cellular staining in the expected compartment and a plausible cell population, such as cytoplasmic staining in kidney tubule cells (HPA: tissue IHC profile; HPA: High in kidney tubule cells). Check the hematoxylin counterstain to distinguish intact cells from necrotic material, and exclude edge staining from scored regions (standard IHC practice). Investigate isolated nuclear-only signal, especially because the reported nucleoplasmic IF location carries a multiple-gene antibody warning (HPA: subcellular summary). A no-primary control helps identify endogenous enzyme or detection background, while a matched positive tissue section tests whether the IHC run produced an interpretable signal (standard IHC practice).
Boster reagents
Best PGAM1 / Phosphoglycerate mutase 1 IHC Antibodies
A04470-2 has IHC data from human paraffin sections and IF/ICC data from HeLa cells (catalog image captions). The catalog lists Human, Mouse, and Rat reactivity (catalog: reactivity).
Real IHC data
Anti-PGAM1 Antibody ®
Cat # A04470-2
A04470-2 was demonstrated by IHC on paraffin sections of human testis cancer and thyroid cancer tissue (A04470-2 IHC captions). Its IF image shows HeLa cells, and its listed applications include IF and ICC (A04470-2 IF caption; catalog: applications).
Which to pick: Choose A04470-2 for chromogenic tissue IHC: its paraffin-section captions specify EDTA retrieval at pH 8.0, 2 μg/ml primary antibody, and HRP/DAB detection (A04470-2 IHC captions). The same SKU is the IF/ICC choice based on its HeLa-cell IF image and listed 5 μg/ml IF concentration (A04470-2 IF caption; catalog: IF dilution). For cross-species work, the catalog lists Human, Mouse, and Rat reactivity, while the supplied images document human material; the IHC captions do not report the fixative (catalog: reactivity; A04470-2 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P18669 (PGAM1_HUMAN, Phosphoglycerate mutase 1).
- Human Protein Atlas. PGAM1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PGAM1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the mid piece. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. PGAM1 antibody validation summary (2 antibodies).
- Phosphoglycerate mutase 1 is highly expressed in C6 glioma cells and human astrocytoma. Oncology letters 2018 — PMC5958715.
- Quantitative proteomics identification of phosphoglycerate mutase 1 as a novel therapeutic target in hepatocellular carcinoma. Molecular cancer 2010 — PMC2873438.
- Phosphoglycerate mutase 1 knockdown inhibits prostate cancer cell growth, migration, and invasion. Asian journal of andrology 2018 — PMC5858104.
- Phosphoglycerate Mutase 1 Predicts the Poor Prognosis of Oral Squamous Cell Carcinoma and is Associated with Cell Migration. Journal of Cancer 2017 — PMC5559954.
- PubMed PMID:2846553 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:2846554 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PGAM1 (Phosphoglycerate mutase 1). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04470-2). Validated IHC antibody: A04470-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human testis cancer tissue; fixative not specified (datasheet A04470-2); 2. Fixation: Image fixative and duration unreported (datasheet A04470-2); 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 A04470-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04470-2); 8. Primary antibody: Rabbit anti-PGAM1, 2-5 μg/ml (datasheet A04470-2); 9. Primary incubation: Overnight at 4 °C (datasheet A04470-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04470-2); Caveat: Possible cross-gene antibody staining (HPA tissue IHC); Isoform / epitope: 0 isoforms annotated; chain spans residues 2–254 (UniProt); Regulation: No expression regulator specified (UniProt). Evidence: UniProt P18669, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A04470-2, PMC5958715, PMC2873438. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PGAM1/immunohistochemistry-ihc-guide.