FOLH1 / Glutamate carboxypeptidase 2 · IHC design guide

Design Immunohistochemistry for FOLH1

Plan paraffin-section IHC for FOLH1 using prostate glands, kidney proximal tubules and intestinal enterocytes as positive tissues (HPA tissue IHC). Interpret the observed cytoplasmic staining (HPA tissue IHC) alongside the annotated membrane topology and isoforms (UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for FOLH1 (IHC for FOLH1): expected localisation Cytoplasmic staining in tissue sections (HPA tissue IHC), antibody M02846-9, validated IHC image, and IHC protocol steps
Printable FOLH1 IHC protocol sheet — expected localisation Cytoplasmic staining in tissue sections (HPA tissue IHC), antibody M02846-9, controls and protocol steps. Open the full FOLH1 IHC guide →

FOLH1 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 tissue sections (HPA tissue IHC)
Staining pattern Cytoplasm of prostate glands, kidney tubules and enterocytes (HPA tissue IHC)
Antigen retrieval Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen)
Positive control ⓘ Duodenum+3 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 sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet M02846-9); verify before use.
Caveat Normal and primary tumor prostate favor different isoforms (UniProt)
Regulation Primary prostate tumors favor PSMA-1 (UniProt)
Isoform / epitope 8 isoforms; check whether the epitope is extracellular or cytoplasmic (UniProt)
Section 1

Recommended FOLH1 IHC & IF Protocols

The catalog antibody protocol and the published ileal IHC protocol provide starting points for FOLH1 staining in paraffin sections (datasheet; PMC3351422 methods).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleTissue sections; selected-image fixative not specified (standard IHC workflow)
FixationImage fixative and duration unreported (datasheet M02846-9); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibodyRabbit anti-FOLH1, 1:200-1:1000 (datasheet M02846-9)
Primary incubationOvernight at 4 °C (standard)
DetectionHRP-polymer secondary, DAB chromogen 5–10 min (standard)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultFOLH1-positive staining in enterocytes of duodenum (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in prostate, kidney tubules, duodenum and small intestine. No signal in the no-primary control.
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); the published ileal protocol used Diva DECLOAKER reagent (PMC3351422 methods).
Section 2

What Is the Expected FOLH1 Staining Pattern?

FOLH1 is a membrane protein with an extracellular region and a short cytoplasmic tail (UniProt Q04609 topology). In paraffin-section IHC, expect strong staining in prostate glandular cells, kidney proximal-tubule microvilli, and duodenal and small-intestinal enterocytes (HPA tissue IHC: High). HPA also describes selective cytoplasmic expression in these tissues and rates the tissue staining profile Enhanced for consistency with RNA expression (HPA tissue IHC).

What am I looking at on my slide?
Strong staining in prostate glandular cells, proximal-tubule microvilli, or intestinal enterocytes.This matches the reported high-expression cell types (HPA tissue IHC: High). Record the stained cell type and whether signal appears at an apical edge, membrane, or within cytoplasm; HPA reports cytoplasmic IHC staining, while UniProt assigns FOLH1 to the membrane and cytoplasm (HPA tissue IHC; UniProt Q04609 subcellular location).
Predominantly nuclear staining in the expected positive cells.Nuclear localisation falls outside the reported membrane and cytoplasmic locations (UniProt Q04609 subcellular location; HPA subcellular: Membrane). Treat a nuclear-only pattern as suspect and check antibody specificity and detection controls before scoring it as FOLH1 (general IHC practice).
Strong staining in a cell type reported as undetected, such as adipocytes.Adipocytes are reported as Not detected (HPA tissue IHC: adipocytes). Consider cross-reactivity or endogenous detection activity, then compare with a no-primary control and an IHC-validated antibody (general IHC practice; HPA antibodies: IHC Enhanced). A single unexpected stain does not establish new FOLH1 expression.
Uniform colour across cells, stroma, or tissue edges, obscuring cell boundaries.This is less interpretable than the selective cell pattern reported by HPA (HPA tissue IHC). Assess nonspecific background with a no-primary control; review blocking, wash steps, and chromogen development under the chosen IHC workflow (general IHC practice).
No staining in prostate glandular cells or the other reported high-expression cells.A blank known-positive section conflicts with the expected high staining (HPA tissue IHC: prostate, kidney, duodenum, small intestine). Check section integrity and run conditions before calling the specimen negative; a failed positive control cannot establish FOLH1 absence (general IHC practice).
💡Expected FOLH1 appearanceCall a result positive when strong staining is selective for prostate glandular cells, kidney proximal-tubule microvilli, or intestinal enterocytes (HPA tissue IHC: High); diffuse tissue-wide or nuclear-only colour is suspect (HPA tissue IHC; UniProt Q04609 subcellular location; general IHC practice).
How each factor affects the staining
Which compartment should be scored?FOLH1 has a cytoplasmic segment at residues 1–19, a transmembrane segment at 20–43, and an extracellular region at 44–750 (UniProt Q04609 topology). HPA calls the tissue IHC pattern cytoplasmic and the subcellular summary membrane (HPA tissue IHC; HPA subcellular). Assess compartment together with cell identity.
Which tissues provide comparison sections?Prostate glandular cells, kidney proximal-tubule microvilli, and duodenal or small-intestinal enterocytes are High (HPA tissue IHC). Adipocytes are Not detected, while salivary glandular cells and hepatocytes are Low (HPA tissue IHC). Use the same staining run for comparisons (general IHC practice).
Can a tissue labelled negative rule out all expression?No. Adrenal glandular cells are Not detected by HPA tissue IHC, while UniProt reports FOLH1 protein detected in adrenal gland (HPA tissue IHC; UniProt Q04609 tissue specificity). Interpret an adrenal result in its assay and cell-type context; these sources do not give a target-specific fixation explanation.
Do isoforms change the expected IHC call?UniProt lists 8 isoforms and reports PSMA' as most abundant in normal prostate, PSMA-1 in primary prostate tumors, and PSMA-9 specifically in prostate cancer (UniProt Q04609 isoforms; tissue specificity). The payload does not identify this antibody's epitope or establish isoform-specific IHC staining.
How strong is the supplied antibody evidence?Both listed antibodies, HPA010593 and CAB001451, have IHC Enhanced status (HPA antibodies). This supports the reported IHC pattern through independent antibodies or orthogonal data (HPA antibodies: IHC Enhanced); it does not validate every specimen or identify the catalog antibody's epitope.
What should the IF/ICC result look like?HPA summarizes FOLH1 as Membrane in its subcellular entry, but provides no main location or ICC-IF image cell lines here (HPA subcellular). UniProt also lists cytoplasm (UniProt Q04609 subcellular location). Treat this as localisation context for the separate IF/ICC guide, without inferring an IF protocol or validated image pattern.
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
The prostate positive control is blank.The section or staining run may have failed; HPA reports High staining in prostate glandular cells (HPA tissue IHC).Repeat with a suitable positive section and verify antibody dilution, retrieval, detection reagents, and counterstain against the chosen IHC protocol (general IHC practice). Do not score test sections as negative until the positive control works.
Staining appears mainly in nuclei.A nuclear-only pattern conflicts with the reported membrane and cytoplasmic locations (UniProt Q04609 subcellular location; HPA subcellular).Inspect the no-primary control, review chromogen deposits and counterstain, and compare the result with an IHC-validated antibody if available (general IHC practice; HPA antibodies: IHC Enhanced).
Many unrelated cells stain as strongly as the expected cells.The broad pattern differs from HPA's selective cytoplasmic tissue profile and may reflect nonspecific staining or endogenous detection activity (HPA tissue IHC; general IHC practice).Compare positive and reported undetected cell types on the same run; check a no-primary control, blocking, and detection steps before assigning FOLH1 positivity (HPA tissue IHC; general IHC practice).
Weak, widespread colour obscures proximal-tubule or enterocyte staining.Background can mask the reported High signal in proximal-tubule microvilli and enterocytes (HPA tissue IHC; general IHC practice).Use the no-primary control to locate background; review washes, antibody dilution, and chromogen development under the chosen IHC protocol (general IHC practice). Reassess the expected cell boundaries afterward (HPA tissue IHC).
Adrenal glandular cells stain despite an HPA Not detected entry.The sources differ in scope: HPA reports those cells Not detected by tissue IHC, while UniProt reports adrenal protein detection (HPA tissue IHC; UniProt Q04609 tissue specificity).Document the cell type and controls, then compare with a reported High tissue and an independent IHC-validated antibody where available (HPA tissue IHC; HPA antibodies: IHC Enhanced). Avoid assigning a fixation cause from these records.
An IF/ICC image seems cytoplasmic rather than sharply membranous.The supplied IF record has a Membrane summary but no ICC-IF image cell lines or main-location detail; UniProt also lists cytoplasm (HPA subcellular; UniProt Q04609 subcellular location).Use the separate IF/ICC guide to assess its own controls and imaging conditions (general IF practice). Do not treat the paraffin IHC tissue pattern or IHC antibody status as direct IF validation (HPA tissue IHC; HPA antibodies).

Sample controls for FOLH1 IHC & IF

🧪Run prostate first: glandular cells should stain (HPA: High in prostate glandular cells). Use adipose tissue as the negative, with adipocytes at background (HPA: Not detected in adipocytes); on the prostate slide, assess nonglandular stroma for background signal against the no-primary control (HPA: High is assigned to glandular cells; standard IHC practice).
Positive control tissue: Duodenum (Enterocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for FOLH1; derive a cell-line control from the positive tissue's cell type (Enterocytes) and confirm it by RNA or western blot first.
Technical controls: Include no-primary and secondary-only controls, a rabbit IgG control matched to the catalog antibody’s isotype and clonality where applicable, and FOLH1-knockout material as a biological negative (caption: rabbit antibody; standard IHC practice). For chromogenic prostate IHC, block and check endogenous peroxidase activity before interpreting signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected M02846-9 prostate caption does not state a fixative (caption: fixative not stated). Retrieval dependence and whether frozen sections or IF would be easier are also unreported; optimize retrieval empirically for paraffin IHC (standard IHC practice). In prostate glands, distinguish cell-associated staining from chromogen trapped in gland lumina using the negative controls (HPA: High in prostate glandular cells; standard IHC practice).

HPA tissue IHC evidence for FOLH1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Duodenum Enterocytes High Protein (IHC) HPA →
Kidney Proximal tubules (microvilli) High Protein (IHC) HPA →
Prostate Glandular cells High Protein (IHC) HPA →
Small intestine Enterocytes High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes Not detected Protein (IHC) HPA →
Adrenal gland Glandular cells Not detected Protein (IHC) HPA →
Appendix Endocrine cells Not detected Protein (IHC) HPA →
Bone marrow Hematopoietic cells Not detected Protein (IHC) HPA →
Bronchus Respiratory epithelial cells Not detected Protein (IHC) HPA →
Section 3

Advanced FOLH1 IHC Tips

These checks help troubleshoot FOLH1 staining in paraffin sections while keeping observed tissue patterns distinct from antibody validation and processing evidence.

What retrieval should I try first when FOLH1 staining is weak?
Use citrate pH 6.0 heat induced epitope retrieval at 95–98 °C for 20 min before chromogenic FOLH1 IHC (page retrieval setting). Compare retrieved and unretrieved sections from the same block, holding antibody incubation and detection constant, because section processing can change epitope accessibility (standard IHC practice). Assess glandular prostate cells and kidney proximal tubule microvilli as expected positive patterns (HPA: High in prostate glandular cells and kidney proximal tubules). If staining remains weak, test another retrieval buffer or pH as a controlled fallback, recording tissue integrity and background alongside signal (standard IHC practice). Do not treat a stronger diffuse signal alone as successful retrieval; evaluate cellular location and a negative control together (standard IHC practice).
Could fixation explain inconsistent FOLH1 staining between paraffin blocks?
Target specific sensitivity to fixative choice or fixation time is unknown here; the selected prostate IHC caption does not state its fixative (caption: M02846-9). Record fixative, duration, tissue thickness and processing history for each block before comparing staining intensity (standard IHC practice). Compare sections from differently processed blocks in one staining run, using the same citrate pH 6.0, 20 min retrieval setting and detection conditions (page retrieval setting; standard IHC practice). Check tissue morphology and an expected positive cell population before assigning weak signal to fixation (standard IHC practice; HPA: High in prostate glandular cells). HPA tissue patterns and FOLH1 topology do not establish a fixation effect for this antibody (HPA: tissue IHC profile; UniProt Q04609 topology).
Should FOLH1 appear membranous or cytoplasmic in prostate sections?
FOLH1 has a membrane segment at residues 20–43, a cytoplasmic segment at 1–19 and an extracellular region at 44–750 (UniProt Q04609 topology). Its annotated locations include cell membrane and cytoplasm, while the tissue IHC profile describes selective cytoplasmic staining (UniProt Q04609 subcellular location; HPA: tissue IHC profile). In prostate, inspect glandular cells for a reproducible cellular pattern and compare it with the High reference staining, rather than requiring a crisp membrane rim in every section (HPA: prostate glandular cells; standard IHC practice). Use morphology and matched controls to distinguish real intracellular signal from diffuse chromogen deposits (standard IHC practice). Record membrane associated and cytoplasmic staining separately when scoring (standard IHC practice).
Can epitope position or splice isoforms change what this antibody detects?
FOLH1 has 8 listed isoforms, and their relative abundance differs between normal prostate and primary prostate tumors (UniProt Q04609 isoforms and tissue specificity). PSMA' is most abundant in normal prostate, whereas PSMA-1 is most abundant in primary prostate tumors; PSMA-9 is described as prostate cancer specific (UniProt Q04609 tissue specificity). The record places residues 1–19 inside the cell and 44–750 outside it, with 10 annotated glycosylation sites in the latter region (UniProt Q04609 topology and glycosylation). Check the antibody's documented immunogen or mapped epitope before claiming isoform coverage or epitope accessibility, because neither is established by the supplied caption (caption: M02846-9; standard IHC practice). Interpret staining differences with the documented epitope and tissue morphology in view (standard IHC practice).
How should I adapt FOLH1 localisation checks for multiplex IF?
Treat multiplex IF as a separate assay: the supplied product evidence is a prostate tissue IHC caption, and the HPA subcellular payload lists no ICC/IF images (caption: M02846-9; HPA: subcellular payload). Pair FOLH1 with a validated marker identifying the expected glandular or enterocyte population, and confirm that each channel preserves its expected cell pattern (HPA: High in prostate glandular cells and duodenal enterocytes; standard IF practice). Choose a fluorophore whose emission is distinguishable from measured tissue autofluorescence, then include single stain and no primary controls when setting exposure and assessing bleed through (standard IF practice). Set permeabilisation according to the documented antibody epitope: access to residues 1–19 differs from access to the extracellular region 44–750 (UniProt Q04609 topology; standard IF practice).
What should I check when chromogen appears across the whole section?
Compare the test section with a no primary control and an expected low signal area before changing antibody concentration (standard IHC practice). Block endogenous peroxidase for a peroxidase and DAB workflow, and inspect whether residual colour follows blood rich or damaged areas rather than intact target cells (standard chromogenic IHC practice). Optimise blocking, washes and detection exposure one variable at a time, keeping the citrate pH 6.0, 20 min retrieval condition fixed during that comparison (page retrieval setting; standard IHC practice). A credible FOLH1 pattern should be selective, consistent with glandular prostate cells or proximal tubule microvilli, rather than uniform staining of every cell (HPA: tissue IHC profile and positive cells). Record background separately from target signal (standard IHC practice).
How can I score FOLH1 staining reproducibly across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and eligible cells before scoring, using glandular prostate cells or kidney proximal tubules when those tissues are examined (HPA: High in prostate glandular cells and kidney proximal tubules; standard IHC practice). For cell based analysis, report percent positive cells and an H-score from intensity categories 0–3, with a possible total of 0–300 (standard IHC scoring practice). Record membrane associated and cytoplasmic signal separately because both locations are annotated and the HPA tissue profile emphasizes cytoplasmic expression (UniProt Q04609 subcellular location; HPA: tissue IHC profile). Normalise positive cell counts to the number of eligible cells, or staining area to the measured viable tissue area in mm², and exclude folds and necrosis using predefined rules (standard IHC practice).
How do I distinguish true FOLH1 positivity from staining artefact?
Look for reproducible cellular staining in a relevant compartment and an expected cell type, such as prostate glandular cells, intestinal enterocytes or kidney proximal tubules (UniProt Q04609 subcellular location; HPA: positive cells). Diffuse colour in unrelated cells, abrupt section edge staining, or deposits over necrosis should trigger review of morphology and controls before interpretation (standard IHC practice). Compare a no primary control to identify detection background, including residual endogenous peroxidase signal in a peroxidase and DAB assay (standard chromogenic IHC practice). HPA reports adipocytes as Not detected, which can provide a tissue specific comparison when adipose tissue is present on a section (HPA: adipose adipocytes). Concordance with an expected pattern supports interpretation but does not establish antibody specificity by itself (standard IHC practice).
Boster reagents

Best FOLH1 / Glutamate carboxypeptidase 2 IHC Antibodies

Catalog anti-FOLH1 antibodies have human prostate IHC images (catalog IHC captions); M02846-9 also lists IF and human, mouse, and rat reactivity (catalog applications/reactivity).

Real IHC data Human prostate was stained with anti-Prostate Specific Membrane Antigen (PSMA) rabbit antibody
Anti-Prostate Specific Membrane Antigen Rabbit Monoclonal Antibody
Cat # M02846-9
Real IHC data Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using PSMA Antibody.
Anti-PSMA FOLH1 Rabbit Monoclonal Antibody
Cat # M02846

M02846-9 will render with an IHC image of human prostate; its listed applications include IHC and IF, with human, mouse, and rat reactivity (catalog IHC caption; catalog applications/reactivity). M02846 will render with an IHC image of paraffin-embedded human prostate carcinoma; it lists IHC and human reactivity (catalog IHC caption; catalog applications/reactivity).

Which to pick: For paraffin-section tissue IHC, choose M02846: its own image caption specifies paraffin-embedded human prostate carcinoma, while the fixative is unreported (catalog IHC caption); its listed IHC dilution is 1:50 (catalog dilution). For IF/ICC, choose M02846-9 because IF is a listed application at 1:200–1:1000 (catalog applications/dilution); its own IHC caption says human prostate but does not report section processing or fixative (catalog IHC caption). For cross-species work, M02846-9 lists human, mouse, and rat reactivity and is described as a recombinant rabbit monoclonal antibody (catalog reactivity/description).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry Q04609 (FOLH1_HUMAN, Glutamate carboxypeptidase 2).
  2. Human Protein Atlas. FOLH1 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. FOLH1 subcellular location (ICC-IF): Membrane.
  4. Human Protein Atlas. FOLH1 antibody validation summary (2 antibodies).
  5. An ileal Crohn's disease gene signature based on whole human genome expression profiles of disease unaffected ileal mucosal biopsies. PloS one 2012 — PMC3351422.
  6. Folate hydrolase-1 (FOLH1) is a novel target for antibody-based brachytherapy in Merkel cell carcinoma. Skin health and disease 2021 — PMC8447486.
  7. Prostate-specific membrane antigen (PSMA) as a potential target for molecular imaging and treatment in bone and soft tissue sarcomas. The British journal of radiology 2023 — PMC10161918.
  8. Pan-cancer analysis reveals FOLH1 as a prognostic biomarker and immune modulator in human tumors. Translational cancer research 2026 — PMC13066985.
  9. PubMed PMID:8417812 — UniProt-cited evidence.
  10. PubMed PMID:7882349 — UniProt-cited evidence.
  11. PubMed PMID:9838072 — UniProt-cited evidence.