This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
AHCYL1 / S-adenosylhomocysteine hydrolase-like protein 1 · IHC design guide
Design Immunohistochemistry for AHCYL1
Plan AHCYL1 staining in paraffin sections using the catalog antibody at 2–5 μg/ml (datasheet A08908-2). Compare cytoplasmic staining with the reported tissue profile, while accounting for its low concordance with RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AHCYL1 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 tissue types (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic; medium in selected glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08908-2) | |
| 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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08908-2) | |
| Caveat | Staining has low concordance with RNA data (HPA tissue IHC) | |
| Regulation | Apoptotic stress reduces MAM localisation (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unreported (UniProt) |
Section 1
Recommended AHCYL1 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet A08908-2); one published AHCYL1 IHC protocol uses sodium citrate retrieval (PMC12068309 methods).
| Sample | Paraffin-embedded human bladder adenosquamous carcinoma tissue; fixative not specified (datasheet A08908-2) |
| Fixation | Image fixative and duration unreported (datasheet A08908-2); 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 A08908-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08908-2) |
| Primary antibody | Rabbit anti-AHCYL1, 2-5 μg/ml (datasheet A08908-2) |
| Primary incubation | Overnight at 4 °C (datasheet A08908-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A08908-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AHCYL1-positive staining in glandular cells of duodenum (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissue types. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet A08908-2). Use the published sodium citrate condition when reproducing the article’s protocol (PMC12068309 methods).
Section 2
What Is the Expected AHCYL1 Staining Pattern?
AHCYL1 is mainly cytosolic, with reported ER and context-dependent apical membrane association; it has no transmembrane segment (UniProt O43865 subcellular location and topology). In paraffin IHC, expect cytoplasmic staining in many tissues, including Medium staining in selected glandular cells (HPA tissue IHC). HPA rates its tissue staining Approved but reports low consistency with RNA data and pending external verification (HPA tissue IHC).
| Cytoplasmic staining in duodenal, endometrial, fallopian tube, pancreatic exocrine or thyroid glandular cells. | This fits the reported Medium staining in those specific cell populations (HPA tissue IHC). Judge the signal against adjacent structures on the same section; the HPA rating does not make every stained cell independently verified (HPA tissue IHC). |
| Predominantly nuclear or luminal staining, with little cytoplasmic signal. | That pattern conflicts with the reported cytosolic localisation (HPA subcellular ICC-IF; UniProt O43865 subcellular location). Review morphology, counterstain and negative controls before calling it AHCYL1; a limited apical signal may be plausible in an appropriate epithelial context (UniProt O43865 subcellular location). |
| Strong signal in a cell population listed as not detected by HPA, such as adipocytes or cardiomyocytes. | Treat the mismatch as a specificity or detection question, including possible cross-reactivity or endogenous chromogen activity (HPA tissue IHC; general IHC practice). A single HPA 'Not detected' entry is a reference observation, not proof that every specimen must be negative (HPA tissue IHC). |
| Broad, even chromogen deposition across cells and empty tissue spaces. | A pattern that ignores cell boundaries is more consistent with background than the reported cytoplasmic cellular profile (HPA tissue IHC; general IHC practice). Check a no-primary control and inspect whether the deposit follows tissue edges or processing artifacts (general IHC practice). |
| No convincing signal in a well-preserved glandular region expected to show Medium staining. | This is discordant with the listed positive populations, but HPA reports limited agreement with RNA data (HPA tissue IHC). First check that the correct cell population is present; then assess IHC controls and detection conditions before interpreting the section as biologically negative (general IHC practice). |
💡Expected AHCYL1 appearanceCall a result positive when glandular cells show interpretable cytoplasmic Medium staining (HPA tissue IHC); nuclear-only, luminal-only or structure-free diffuse deposit is suspect (HPA subcellular ICC-IF; general IHC practice).
| Tissue and cell selection | HPA reports cytoplasmic expression in most tissue types and Medium staining in five listed glandular populations (HPA tissue IHC). Use the named cells, rather than the tissue name alone, when choosing a reference area (general IHC practice). |
| Antibody evidence | The listed rabbit polyclonal HPA042589 has IHC status Approved (HPA antibodies). Tissue staining is also rated Approved, with low RNA agreement and external verification pending (HPA tissue IHC); this limits how confidently an unexpected pattern can be assigned to AHCYL1. |
| Intracellular distribution | AHCYL1 is cytosolic and ER-associated, lacks a transmembrane segment, and can associate with membranes when phosphorylated (UniProt O43865 subcellular location and topology). Its MAM localisation decreases under apoptotic stress (UniProt O43865, PubMed:27995898); these annotations do not predict a fixed-section staining intensity. |
| Isoforms and antibody coverage | UniProt lists 2 isoforms (UniProt O43865 isoforms). No epitope map or isoform-specific IHC result is supplied, so an observed stain cannot be assigned to one isoform, and a negative stain cannot establish that either isoform is absent. |
| IF/ICC Q: What localisation should be expected? | A: HPA calls cytosol the approved main ICC-IF location and lists images from A-431, U-251MG and U2OS (HPA subcellular ICC-IF). That microscopy evidence helps interpret compartment, but IF/ICC assay design belongs on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclear-only or luminal-only staining dominates. | The compartment does not match the reported cytosolic pattern; luminal pigment or nonspecific staining may contribute (HPA subcellular ICC-IF; general IHC practice). | Compare with a no-primary control, inspect cell boundaries and counterstain, and repeat with an independently validated antibody if available (general IHC practice). |
| Unexpected cells stain strongly while nearby expected glandular cells do not. | Possible cross-reactivity, endogenous detection activity or a cell-identification error; the expected populations are cell-specific (HPA tissue IHC; general IHC practice). | Confirm cell identity on the section, check detection controls and compare a named HPA-positive population under the same IHC conditions (HPA tissue IHC; general IHC practice). |
| Diffuse brown haze obscures cytoplasm. | Background from detection reagents or incomplete blocking can obscure cellular staining (general IHC practice). | Review no-primary and reagent controls; adjust blocking, washes and detection exposure within the assay's validated range (general IHC practice). |
| No signal in an expected Medium glandular population. | The sampled cells may be absent, or retrieval, antibody exposure or detection may have failed (HPA tissue IHC; general IHC practice). | Verify morphology and control performance; review retrieval and antibody conditions without assuming AHCYL1-specific fixation sensitivity, which is unreported in the supplied sources (general IHC practice). |
| Apical accentuation appears alongside cytoplasmic stain. | Context-dependent apical membrane association is annotated, although cytosol is the main HPA ICC-IF location (UniProt O43865 subcellular location; HPA subcellular ICC-IF). | Check that the apical edge belongs to the stained cells and that negative controls are clean; do not require apical staining in every positive section (general IHC practice). |
| A specimen disagrees with the HPA positive or not-detected list. | HPA's tissue IHC rating is Approved but has low consistency with RNA expression and awaits external verification (HPA tissue IHC). | Record cell type, compartment and control results, then seek independent antibody or orthogonal evidence before treating the discrepancy as a new biological finding (general IHC practice). |
Experiment evidence
IHC & IF images showing AHCYL1 staining
A08908-2 · IHCIHC analysis of IRBIT/AHCYL1 using anti-IRBIT/AHCYL1 antibody (A08908-2). IRBIT/AHCYL1 was detected in a paraffin-embedded section of human
A08908-2 · IHCIHC analysis of IRBIT/AHCYL1 using anti-IRBIT/AHCYL1 antibody (A08908-2). IRBIT/AHCYL1 was detected in a paraffin-embedded section of human
A08908-2 · IFIF analysis of IRBIT/AHCYL1 using anti-IRBIT/AHCYL1 antibody (A08908-2). IRBIT/AHCYL1 was detected in an immunocytochemical section of A431
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000168710-AHCYL1/tissue/cerebral+cortex#imid_11261491
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000168710-AHCYL1/tissue/colon#imid_11261433
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000168710-AHCYL1/tissue/liver#imid_11261418
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000168710-AHCYL1/tissue/pancreas#imid_11261385
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042589; green = AHCYL1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042589; green = AHCYL1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AHCYL1 IHC & IF
🧪Run pancreas first and assess its exocrine glandular cells for staining (HPA: Medium in pancreatic exocrine glandular cells). Run adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the pancreas slide, count other cells as internal negatives only after verifying their status, and expect those cells to lack specific staining.
Positive control tissue: Duodenum (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AHCYL1 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality, and AHCYL1-knockout material as a biological negative (selected-SKU caption: rabbit primary antibody). Block endogenous peroxidase and biotin and inspect background before interpreting DAB staining (selected-SKU caption: streptavidin–biotin detection with DAB).
⚠️Feasibility: The selected-SKU paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but does not report the fixative (selected-SKU caption). A target-specific fixation window and whether retrieval is required are unreported; the caption documents one successful retrieval condition, not a comparison. The supplied evidence does not establish whether frozen sections or IF are easier; in pancreas, check exocrine background alongside the staining because the positive population is exocrine glandular cells (HPA: Medium in pancreatic exocrine glandular cells).
HPA tissue IHC evidence for AHCYL1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 | Medium | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Medium | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | Medium | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AHCYL1 IHC Tips
Use the catalog antibody’s paraffin-section result as the starting point, then assess compartment, controls, and tissue context before interpreting AHCYL1 staining.
Which retrieval conditions should I try first for AHCYL1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A08908-2). The catalog antibody stained a paraffin-embedded human bladder adenosquamous carcinoma section after that treatment, with 2 μg/ml primary antibody incubated overnight at 4°C (datasheet A08908-2). If staining is weak, compare retrieval duration on adjacent sections while keeping antibody concentration, detection, and exposure to DAB consistent (standard IHC practice). A citrate buffer at pH 6.0 is a fallback to test only after the documented EDTA condition; judge gains against tissue damage and background (standard IHC practice).
Could fixation explain weak or uneven AHCYL1 staining?
The selected tissue-IHC caption identifies a paraffin-embedded section but does not report its fixative, so AHCYL1-specific fixation sensitivity is unknown (datasheet A08908-2). Record fixation method and duration for each specimen, and compare sections processed together when investigating weak or uneven staining (standard IHC practice). Keep the documented EDTA retrieval at pH 8.0 and the primary incubation at 2 μg/ml overnight at 4°C constant during that comparison (datasheet A08908-2). Examine morphology and staining across the section before attributing a difference to fixation; neither the reported tissue pattern nor AHCYL1 topology establishes a fixation effect (HPA: tissue IHC; UniProt O43865 topology).
Where should convincing AHCYL1 staining appear within cells?
Assess cytoplasmic staining first: tissue IHC reports cytoplasmic expression in most tissue types, and the approved subcellular location is cytosol (HPA: tissue IHC; HPA: subcellular). AHCYL1 is also annotated at the endoplasmic reticulum, microsomes, and apical cell membrane, without a transmembrane segment (UniProt O43865 localisation and topology). A fine membrane-associated component can therefore be plausible, but chromogenic light microscopy alone cannot establish localisation at mitochondria-associated ER membranes (UniProt O43865 localisation; standard IHC interpretation). Compare the pattern with cell boundaries and a counterstain, and investigate predominantly nuclear or diffuse extracellular DAB before assigning it to AHCYL1 (standard IHC practice).
Can this IHC result distinguish AHCYL1 isoforms or phosphorylation states?
Do not assign a stained cell to isoform 1 or 2 without an isoform-specific epitope map or independent validation; two isoforms are listed, but no antibody epitope is supplied here (UniProt O43865 isoforms; datasheet A08908-2). AHCYL1 also has annotated modifications, including phosphoserines at residues 68 and 71 (UniProt O43865 modified residues). Retrieval or fixation could affect epitope accessibility in general, yet this evidence does not establish how either modification changes this antibody’s IHC signal (standard IHC practice; datasheet A08908-2). If isoform or phosphostate is the question, compare separately validated reagents on matched sections and report this stain as total detectable AHCYL1 immunoreactivity (standard IHC practice).
How should I adapt the IHC localisation question for multiplex IF?
Treat multiplex IF as a separate assay: the selected antibody evidence describes chromogenic staining of a paraffin section, not an IF protocol (datasheet A08908-2). Pair AHCYL1 with a validated marker of the cell population being assessed, such as a glandular-cell marker when examining an HPA-positive glandular compartment (HPA: tissue IHC; standard IF practice). Choose fluorophores after checking the tissue’s autofluorescence, favoring a spectrally separated, longer-wavelength channel for a weak AHCYL1 signal when appropriate (standard IF practice). Because AHCYL1 is reported in cytosol and at membrane-associated sites without a transmembrane segment, optimise permeabilisation against the antibody’s mapped epitope, if known, and check that cell architecture remains intact (HPA: subcellular; UniProt O43865 topology; standard IF practice).
What should I check when DAB staining is widespread or granular?
Run a no-primary control and inspect whether diffuse DAB persists through the detection steps; that pattern points toward background from the workflow rather than specific primary binding (standard IHC practice). The documented method uses 10% goat serum block, biotinylated goat anti-rabbit secondary for 30 minutes at 37°C, a streptavidin-biotin complex, and DAB (datasheet A08908-2). Check peroxidase blocking and, with this biotin-based method, assess endogenous biotin as general sources of signal (standard IHC practice). Then titrate the primary around the documented 2 μg/ml condition and compare cytoplasmic cell staining with extracellular deposits or section-edge intensity (datasheet A08908-2; HPA: tissue IHC; standard IHC practice).
How can I score AHCYL1 IHC across heterogeneous samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since the reported tissue-IHC pattern is cytoplasmic and some listed cell populations have no detected staining (HPA: tissue IHC). For a graded signal, record the percentage of cells at intensity 0–3 and calculate an H-score from 0–300; alternatively report percentage positive cells using a prespecified threshold (standard IHC practice). If counting discrete positive cells, report density per mm² of evaluable tissue, with necrosis and folds excluded by consistent rules (standard IHC practice). Normalise comparisons to the same cell type, tissue area, staining batch, and control reference so differences in cellular composition or DAB development do not masquerade as AHCYL1 changes (standard IHC practice).
How do I separate true AHCYL1 signal from artefact?
A credible result has cell-associated cytoplasmic staining in an appropriate compartment, consistent with the tissue-IHC profile and approved cytosolic localisation (HPA: tissue IHC; HPA: subcellular). Medium staining is reported in duodenal and pancreatic glandular populations, whereas adipocytes and several other specified populations were not detected; use these as contextual comparisons, not absolute biological negatives (HPA: tissue IHC). Scrutinise predominantly nuclear signal, staining confined to section edges, necrotic debris, and DAB remaining in a no-primary control (standard IHC practice). Check endogenous peroxidase and biotin-related background for the documented streptavidin-biotin DAB workflow, and interpret any positive result cautiously because HPA notes low staining–RNA consistency and pending external verification (datasheet A08908-2; HPA: reliability; standard IHC practice).
Boster reagents
Best AHCYL1 / S-adenosylhomocysteine hydrolase-like protein 1 IHC Antibodies
The IHC-validated antibody A08908-2 has human paraffin-section IHC images and A431-cell IF data (catalog figure captions); the catalog lists Human, Mouse and Rat reactivity (catalog reactivity).
Real IHC data
Anti-IRBIT/AHCYL1 Antibody ®
Cat # A08908-2
A08908-2 is shown staining paraffin-embedded human bladder adenosquamous carcinoma, Hashimoto thyroiditis, thyroid papillary carcinoma and laryngeal squamous cell carcinoma sections (IHC image captions). The same SKU is shown in A431-cell IF and is listed for both ICC and IF (IF image caption; catalog applications).
Which to pick: For tissue IHC, choose A08908-2: its own captions document paraffin sections stained at 2 μg/ml, but do not report the fixative (IHC image captions). For IF/ICC, the same SKU has an A431-cell IF image at 5 μg/ml; its clone is unspecified (IF image caption; catalog clone field). For Mouse or Rat work, the same SKU lists those species as reactive, although the supplied IHC captions show human tissue and the IF caption shows A431 cells (catalog reactivity; IHC and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43865 (SAHH2_HUMAN, S-adenosylhomocysteine hydrolase-like protein 1).
- Human Protein Atlas. AHCYL1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. AHCYL1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. AHCYL1 antibody validation summary (1 antibodies).
- S-adenosylhomocysteine hydrolase-like protein 1 (AHCYL1) inhibits lung cancer tumorigenesis by regulating cell plasticity. Biology direct 2023 — PMC9985837.
- AHCYL1 Is a Novel Biomarker for Predicting Prognosis and Immunotherapy Response in Colorectal Cancer. Journal of oncology 2022 — PMC9107370.
- AHCYL1 is mediated by estrogen-induced ERK1/2 MAPK cell signaling and microRNA regulation to effect functional aspects of the avian oviduct. PloS one 2012 — PMC3492294.
- AHCYL1 mediates the tumor-promoting effect of PREX2 in non-small cell lung carcinoma. Theranostics 2025 — PMC12068309.
- PubMed PMID:11904675 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AHCYL1 (S-adenosylhomocysteine hydrolase-like protein 1). Expected localisation Cytoplasmic staining in most tissue types (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08908-2). Validated IHC antibody: A08908-2 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 bladder adenosquamous carcinoma tissue; fixative not specified (datasheet A08908-2); 2. Fixation: Image fixative and duration unreported (datasheet A08908-2); 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 A08908-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08908-2); 8. Primary antibody: Rabbit anti-AHCYL1, 2-5 μg/ml (datasheet A08908-2); 9. Primary incubation: Overnight at 4 °C (datasheet A08908-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08908-2); Caveat: Staining has low concordance with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unreported (UniProt); Regulation: Apoptotic stress reduces MAM localisation (UniProt). Evidence: UniProt O43865, HPA tissue IHC · Approved, HPA ICC-IF · cytosol., Datasheet A08908-2, PMC12068309. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AHCYL1/immunohistochemistry-ihc-guide.