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SUMF2 / Inactive C-alpha-formylglycine-generating enzyme 2 · IHC design guide
Design Immunohistochemistry for SUMF2
Plan chromogenic SUMF2 IHC in paraffin sections using cytoplasmic staining as the tissue readout (HPA tissue IHC). Start the catalog antibody at 2–5 μg/ml (datasheet A08890), with kidney proximal tubules as a high-staining control and adipocytes as a non-detected comparator (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SUMF2 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 (HPA tissue IHC); ER lumen expected (UniProt) | |
| Staining pattern | General cytoplasmic staining in positive cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08890) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across test and control paraffin sections. (selected-SKU IHC image A08890) | |
| Caveat | Staining has medium agreement with RNA data; verify specificity (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 5 isoforms; mature chain 26–301; epitope unspecified (UniProt) |
Section 1
Recommended SUMF2 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A08890). Two published SUMF2 IHC protocols provide additional conditions (PMC11298345; PMC11803025).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08890) |
| Fixation | Image fixative and duration unreported (datasheet A08890); 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 A08890); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08890) |
| Primary antibody | Rabbit anti-SUMF2, 2-5 μg/ml (datasheet A08890) |
| Primary incubation | Overnight at 4 °C (datasheet A08890) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08890) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SUMF2-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 retrieval for the catalog antibody (datasheet A08890); the published protocols use citrate pH 6.0 (PMC11298345) and Tris-EDTA pH 9.0 (PMC11803025).
Section 2
What Is the Expected SUMF2 Staining Pattern?
SUMF2 is a soluble endoplasmic reticulum lumen protein with no transmembrane segment (UniProt Q8NBJ7 topology). In paraffin section IHC, expect mainly cytoplasmic staining, especially in glandular cells, kidney proximal tubule microvilli, pancreatic endocrine cells, and placental syncytiotrophoblast cell bodies (HPA tissue IHC). HPA rates the tissue pattern Enhanced, while reporting medium agreement with RNA data and pending external verification (HPA tissue IHC).
| Strong cytoplasmic staining in duodenal, endometrial, gallbladder, seminal vesicle, or small intestinal glandular cells. | This matches HPA's High glandular cell staining and general cytoplasmic pattern (HPA tissue IHC). Score the named cell population and its cytoplasm; the UniProt ER lumen assignment supports an intracellular pattern but does not define a specific chromogenic texture (UniProt Q8NBJ7). |
| Strong signal at kidney proximal tubule microvilli, in pancreatic endocrine cells, or in syncytiotrophoblast cell bodies. | These are separate High HPA tissue observations (HPA tissue IHC). Record the precise compartment described for each tissue; do not interpret the kidney microvillar observation as evidence that SUMF2 is generally a plasma membrane protein (HPA tissue IHC; UniProt Q8NBJ7 topology). |
| Predominantly nuclear, extracellular, or broadly surface outlining stain replaces the expected cellular pattern. | Treat this as a localisation mismatch requiring control review: UniProt places SUMF2 in the ER lumen without a transmembrane segment, whereas HPA tissue IHC describes general cytoplasmic expression (UniProt Q8NBJ7 topology; HPA tissue IHC). A mismatch alone does not identify its cause. |
| Strong staining appears in adipocytes, cardiomyocytes, or another cell population HPA reports as not detected. | Check for antibody cross-reactivity or endogenous chromogen detection activity before calling the tissue positive (general IHC practice). HPA reports Not detected in those specified cells, but its Enhanced tissue profile still awaits external verification (HPA tissue IHC). |
| The whole section is diffusely colored, or a known HPA high staining cell population shows no signal. | Diffuse color prevents cell specific scoring; absent signal in an expected positive population makes a negative call unreliable (general IHC practice). Compare a matched negative reagent control and an HPA high tissue control before changing the biological interpretation (HPA tissue IHC; general IHC practice). |
💡Expected SUMF2 appearanceCall a result positive when chromogenic signal is predominantly cytoplasmic in an HPA High cell population, with the reported kidney microvillar pattern assessed separately; nuclear, extracellular, or uniform staining across HPA not detected populations is suspect (HPA tissue IHC; UniProt Q8NBJ7 topology).
| Cell and tissue selection | HPA reports High staining in several glandular populations, proximal tubule microvilli, endocrine pancreas, and syncytiotrophoblast cell bodies, but Not detected in specified adipocytes and cardiomyocytes (HPA tissue IHC). Choose controls by cell population, not merely by organ name. |
| Protein localisation and processing | UniProt assigns SUMF2 to the ER lumen, lists a signal peptide at residues 1–25, and identifies the mature chain as residues 26–301 (UniProt Q8NBJ7). The supplied record gives no antibody epitope, so these annotations cannot predict whether a particular processed region is detected. |
| Isoforms and glycosylation | UniProt lists 5 isoforms and one glycosylation site at residue 191 (UniProt Q8NBJ7). Without an antibody epitope or isoform coverage statement, neither annotation establishes a different IHC pattern or a required antigen retrieval condition. |
| Evidence and antibody choice | HPA rates tissue IHC Enhanced but notes medium agreement with RNA data and pending external verification (HPA tissue IHC). HPA024040 and CAB025743 each have Enhanced IHC validation; the supplied record does not establish that either catalog antibody shares their epitope or performance (HPA antibodies). |
| IF/ICC Q: Should its localisation determine this IHC call? | A: HPA lists vesicles and plasma membrane as uncertain ICC-IF locations, with images in A-431, SiHa, and U2OS (HPA subcellular). Use the tissue IHC pattern for paraffin section scoring; reconcile discordant compartments with the UniProt ER lumen annotation (HPA tissue IHC; UniProt Q8NBJ7). |
| Detection chemistry and fixation evidence | Endogenous detection activity can create color unrelated to antibody binding, so assess the chosen chromogenic system with appropriate controls (general IHC practice). No target specific fixation sensitivity, retrieval condition, or dilution is supplied; avoid attributing a SUMF2 result to any of these as an established target effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in a glandular cell population reported as High. | Possible failed detection, weak antibody signal, or unsuitable assay conditions; the slide alone cannot distinguish these (general IHC practice). | Run a known positive tissue and the assay controls together; check antibody, secondary detection, chromogen development, and the established retrieval workflow before calling the sample negative (HPA tissue IHC; general IHC practice). |
| Stain covers nearly every cell and the surrounding section. | Diffuse background may reflect nonspecific binding, incomplete blocking, or detection chemistry rather than the HPA cell specific pattern (HPA tissue IHC; general IHC practice). | Inspect the negative reagent control, review blocking and washing, and adjust the working antibody concentration within validated assay limits (general IHC practice). Reassess only after cellular boundaries become interpretable. |
| A mainly nuclear or extracellular pattern dominates. | This conflicts with the ER lumen assignment and HPA general cytoplasmic tissue pattern; its specific cause remains unproven (UniProt Q8NBJ7 topology; HPA tissue IHC). | Compare a matched positive tissue and negative reagent control, then verify the antibody identity and detection setup before scoring the unusual compartment as SUMF2 (general IHC practice). |
| Adipocytes or cardiomyocytes stain as strongly as the expected positive cells. | HPA lists these cell populations as Not detected; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare no primary antibody and positive tissue controls, and localise the color to exact cell types; confirm any persistent unexpected signal independently (general IHC practice). |
| Kidney staining appears restricted to proximal tubule microvilli. | That distribution matches HPA's High microvillar observation, although UniProt assigns SUMF2 to the ER lumen (HPA tissue IHC; UniProt Q8NBJ7). | Document the microvillar distribution separately from broader cytoplasmic staining and compare it with other HPA high cell populations before drawing a general localisation conclusion (HPA tissue IHC). |
| IF/ICC shows vesicular or surface signal that differs from the paraffin section. | HPA calls both ICC-IF locations uncertain; the tissue IHC profile has a separate Enhanced assessment (HPA subcellular; HPA tissue IHC). | Interpret each application against its own controls and evidence; use the tissue IHC pattern for this chromogenic result and document the discrepancy without assigning a new SUMF2 compartment (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing SUMF2 staining
A08890 · IHCIHC analysis of SUMF2 using anti-SUMF2 antibody (A08890). SUMF2 was detected in a paraffin-embedded section of human breast cancer tissue. H
A08890 · IHCIHC analysis of SUMF2 using anti-SUMF2 antibody (A08890). SUMF2 was detected in a paraffin-embedded section of human colon adenocarcinoma ti
A08890 · IFIF analysis of SUMF2 using anti-SUMF2 antibody (A08890). SUMF2 was detected in a paraffin-embedded section of human colon cancer tissue. Hea
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000129103-SUMF2/tissue/cerebral+cortex#imid_6134112
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000129103-SUMF2/tissue/colon#imid_6134038
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000129103-SUMF2/tissue/liver#imid_6134055
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000129103-SUMF2/tissue/kidney#imid_6134056
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA074892; green = SUMF2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA074892; green = SUMF2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SUMF2 IHC & IF
🧪Run duodenum first and expect its glandular cells to stain (HPA: High in duodenal glandular cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); on the duodenum slide, nonglandular cells should show background-level signal, but their SUMF2-negative status is unverified by the supplied HPA rows.
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SUMF2 in A-431, SiHa, U2OS, with annotated localisation: Vesicles (uncertain), Plasma membrane (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; rabbit IgG matched to the primary antibody’s clonality and concentration; and a SUMF2 knockout sample or immunizing-peptide competition if the peptide is available (caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase for HRP/DAB detection in duodenum, and check tissue autofluorescence before interpreting IF signal (caption: HRP/DAB detection; standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A08890 paraffin-section caption does not state a fixative (caption: fixative not stated). That caption documents heat retrieval in EDTA at pH 8.0, so use it as a starting condition; retrieval dependency and whether frozen sections or IF are easier remain unreported (caption: EDTA retrieval). Duodenal background from endogenous peroxidase or autofluorescence should be assessed with the corresponding controls (standard IHC/IF practice).
HPA tissue IHC evidence for SUMF2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Proximal tubules (microvilli) | High | Protein (IHC) | HPA → |
| Pancreas | Pancreatic endocrine cells | High | 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 → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SUMF2 IHC Tips
Use the catalog antibody’s paraffin section evidence as a starting point, then judge SUMF2 staining by compartment, cell type and controls.
What retrieval should I use when SUMF2 staining is weak in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A08890). The selected paraffin section image used that retrieval before incubation with 2 μg/ml antibody overnight at 4°C (A08890 tissue-IHC caption). If signal is weak, first check that sections received consistent heating and cooling, then compare a controlled retrieval adjustment on adjacent sections (standard IHC practice). Keep antibody concentration, incubation and DAB development matched so a retrieval change can be judged independently (standard IHC practice). Score intracellular staining against tissue background, since SUMF2 is assigned to the endoplasmic reticulum lumen (UniProt Q8NBJ7).
Could fixation explain weak or uneven SUMF2 staining?
SUMF2 specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin section caption does not state a fixative (A08890 tissue-IHC caption). Record the fixative and fixation duration for each specimen before comparing staining intensity across sections (standard IHC practice). Process matched control and test sections together, since fixation and subsequent processing can affect antigen accessibility in chromogenic IHC (standard IHC practice). If one specimen stains weakly, review preservation, section quality and retrieval alongside the antibody incubation used in the caption, 2 μg/ml overnight at 4°C (A08890 tissue-IHC caption). Do not assign a SUMF2 specific fixation effect without a direct comparison.
Should SUMF2 appear in the cytoplasm or at the cell surface?
Expect predominantly intracellular staining because SUMF2 is assigned to the endoplasmic reticulum lumen and has no transmembrane segment (UniProt Q8NBJ7 topology). HPA describes general cytoplasmic expression in tissue IHC, while its vesicle and plasma membrane IF locations are marked uncertain (HPA tissue IHC; HPA subcellular). Inspect well preserved cells at high magnification and record whether staining follows cytoplasm, cell borders or extracellular material (standard IHC practice). Compare that pattern with proximal tubules and syncytiotrophoblast cell bodies, both reported as strongly stained by HPA (HPA tissue IHC). Treat a sharp membrane only pattern as needing independent validation, especially when adjacent intracellular compartments show no signal (UniProt Q8NBJ7 topology; standard IHC practice).
How do I troubleshoot discordant staining across SUMF2 isoforms or processing states?
SUMF2 has 5 annotated isoforms, a signal peptide at residues 1–25 and a mature chain at 26–301 (UniProt Q8NBJ7). The record also lists a glycosylation site at residue 191, but the supplied antibody evidence does not map its epitope (UniProt Q8NBJ7; A08890 tissue-IHC caption). Check the immunogen or epitope documentation before attributing different staining patterns to an isoform or processing state (standard IHC practice). Keep retrieval and detection constant while comparing sections, because altered antigen exposure can resemble an epitope dependent difference (standard IHC practice). Report compartment and cell type with intensity; chromogenic staining alone does not identify which SUMF2 isoform was detected.
How can I assess SUMF2 by IF alongside a cell type marker?
For a separate IF experiment, pair SUMF2 with a marker for the expected cell population, such as a validated proximal tubule marker when examining kidney, where HPA reports high proximal tubule staining (HPA tissue IHC). Choose spectrally separated fluorophores and place the weaker channel where tissue autofluorescence is lower, checking unstained and single stain controls (standard IF practice). SUMF2 is assigned to the endoplasmic reticulum lumen, so assess whether permeabilisation allows antibody access to an intracellular epitope (UniProt Q8NBJ7 topology; standard IF practice). Its precise antibody epitope is unspecified, and HPA labels vesicle and plasma membrane IF localisation uncertain (A08890 tissue-IHC caption; HPA subcellular). Interpret colocalisation only after checking bleed through and background in each channel (standard IF practice).
What should I check when SUMF2 DAB staining is diffuse or patchy?
The selected paraffin section workflow used 10% goat serum blocking, a peroxidase conjugated secondary antibody and DAB detection (A08890 tissue-IHC caption). Include a no primary control to assess secondary binding and endogenous chromogenic signal, and block endogenous peroxidase as appropriate for the tissue (standard IHC practice). Inspect section edges, folds and damaged areas separately from intact cells, because these regions can accumulate misleading stain (standard IHC practice). If background persists, compare matched antibody dilutions while holding retrieval and DAB development constant; 2 μg/ml overnight at 4°C is the documented starting incubation (A08890 tissue-IHC caption). Judge specificity against the expected intracellular pattern rather than overall brown intensity (UniProt Q8NBJ7 topology; standard IHC practice).
How should I score SUMF2 staining across mixed cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: HPA reports high staining in kidney proximal tubules and pancreatic endocrine cells, while breast adipocytes are not detected (HPA tissue IHC). For a chromogenic readout, record percentage of positive target cells and an intensity grade of 0–3, then calculate an H-score on the conventional 0–300 scale (standard IHC practice). Alternatively, report positive cell density per mm² when discrete cells can be reliably segmented (standard IHC practice). Normalise to the number or area of the specified cell population and use the same staining run, threshold and exclusion rules across samples (standard IHC practice). Exclude folds, necrosis and section edges before comparing scores (standard IHC practice).
How can I distinguish genuine SUMF2 staining from an IHC artefact?
A plausible result shows cellular staining compatible with SUMF2’s endoplasmic reticulum lumen assignment and the general cytoplasmic tissue pattern reported by HPA (UniProt Q8NBJ7 topology; HPA tissue IHC). Check whether the stained cells match a documented population, such as high staining in duodenal glandular cells or kidney proximal tubules (HPA tissue IHC). A signal confined to nuclei, tissue edges or necrotic material warrants review of localisation and section quality before interpretation (UniProt Q8NBJ7 topology; standard IHC practice). Compare no primary and peroxidase blocked controls to identify detection related colour, particularly where endogenous enzyme activity is possible (standard IHC practice). HPA’s tissue IHC reliability is Enhanced but awaits external verification, so confirm unexpected patterns independently (HPA tissue IHC).
Boster reagents
Best SUMF2 / Inactive C-alpha-formylglycine-generating enzyme 2 IHC Antibodies
A08890 has IHC data from human paraffin breast cancer sections and IF data from human paraffin colon cancer, ovarian cancer and placenta sections (catalog: A08890 image captions).
Real IHC data
Anti-SUMF2 Antibody ®
Cat # A08890
A08890 is a rabbit antibody listed for human IHC and IF (catalog: A08890 host, applications and reactivity). Its captions show IHC in human paraffin breast cancer, colon adenocarcinoma, ovarian tumor and placenta sections, and IF in human paraffin colon cancer, ovarian cancer and placenta sections (catalog: A08890 image captions).
Which to pick: For tissue IHC, choose A08890: its paraffin breast cancer image uses EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, within the listed 2–5 μg/ml IHC range (A08890 IHC image caption; catalog: A08890 IHC dilution). For IF, A08890 is listed at 5 μg/ml and shown in human paraffin sections; the captions do not report a fixative (catalog: A08890 IF dilution and image captions). ICC and cross-species validation are not listed, so this payload does not support a recommendation for either use (catalog: A08890 applications and human reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NBJ7 (SUMF2_HUMAN, Inactive C-alpha-formylglycine-generating enzyme 2).
- Human Protein Atlas. SUMF2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SUMF2 subcellular location (ICC-IF): Localized to the plasma membrane and vesicles..
- Human Protein Atlas. SUMF2 antibody validation summary (3 antibodies).
- Advancing bladder cancer management: development of a prognostic model and personalized therapy. Frontiers in immunology 2024 — PMC11298345.
- Sulfatase modifying factor 2 as a predictive biomarker for urothelial carcinoma. Discover oncology 2025 — PMC11803025.
- PubMed PMID:12757706 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SUMF2 (Inactive C-alpha-formylglycine-generating enzyme 2). Expected localisation Cytoplasmic staining (HPA tissue IHC); ER lumen expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08890). Validated IHC antibody: A08890 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08890); 2. Fixation: Image fixative and duration unreported (datasheet A08890); 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 A08890); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08890); 8. Primary antibody: Rabbit anti-SUMF2, 2-5 μg/ml (datasheet A08890); 9. Primary incubation: Overnight at 4 °C (datasheet A08890). Watch for: Fixation: Keep fixation consistent across test and control paraffin sections. (selected-SKU IHC image A08890); Caveat: Staining has medium agreement with RNA data; verify specificity (HPA tissue IHC); Isoform / epitope: 5 isoforms; mature chain 26–301; epitope unspecified (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt Q8NBJ7, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and vesicles., Datasheet A08890, PMC11298345, PMC11803025. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SUMF2/immunohistochemistry-ihc-guide.