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ADAM23 / Disintegrin and metalloproteinase domain-containing protein 23 · IHC design guide
Design Immunohistochemistry for ADAM23
Plan ADAM23 chromogenic IHC on paraffin sections with an antibody titration of 1:50–1:200 (datasheet: 1:50–1:200). Use nasopharyngeal respiratory epithelial cells or testis Leydig cells as high-staining references, and assess cytoplasmic expression and plasma positivity (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ADAM23 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 expression and plasma positivity (HPA tissue IHC) | |
| Staining pattern | High in respiratory epithelial and Leydig cells; cytoplasmic/plasma (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Nasopharynx+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
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 A07382) | |
| Caveat | A secreted variant may shift RNA and protein tissue patterns (HPA tissue IHC) | |
| Regulation | Brain enriched; heart weak (UniProt) | |
| Isoform / epitope | 3 isoforms; check extracellular versus cytoplasmic epitope (UniProt) |
Section 1
Recommended ADAM23 IHC & IF Protocols
The catalog antibody protocol is accompanied by published ADAM23 IHC methods for paraffin skin sections (PMC10750783) and FFPE tumor sections (PMC6500989).
| Sample | Paraffin-embedded human rectum carcinoma tissue; fixative not specified (datasheet A07382) |
| Fixation | Image fixative and duration unreported (datasheet A07382); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-ADAM23, 1:50-1:200 (datasheet A07382) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ADAM23-positive staining in respiratory epithelial cells of nasopharynx (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression and plasma positivity. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); use 121 °C for 10 min when reproducing the skin protocol (PMC10750783).
Section 2
What Is the Expected ADAM23 Staining Pattern?
ADAM23 is a membrane-associated protein with an extracellular region and a short cytoplasmic tail; UniProt also lists a secreted location (UniProt O75077 topology and location). In tissue IHC, expect cytoplasmic expression and plasma-membrane positivity, particularly in nasopharyngeal respiratory epithelium and testicular Leydig cells (HPA tissue IHC: Approved; High in both cell types).
| Strong cellular staining in nasopharyngeal respiratory epithelium or testicular Leydig cells (HPA: High in both). | This fits the reported positive-cell pattern; assess cytoplasmic and plasma-membrane signal against the surrounding tissue (HPA: tissue IHC profile). |
| A mainly nuclear IHC signal with little cytoplasmic or membrane staining (HPA: tissue IHC profile). | Treat this as discordant with the tissue IHC profile and investigate artefact or antibody specificity; nucleoplasmic staining is separately approved in ICC-IF (HPA: subcellular ICC-IF). |
| Strong staining in cells outside the reported positive populations (HPA: tissue IHC cell-level results). | Check the cell identification and controls before calling it ADAM23; cross-reactivity or endogenous chromogen activity can mimic a positive result (general IHC practice). |
| Uniform colour across cells and extracellular space, with little cell-specific contrast (general IHC practice). | This favours diffuse background over an interpretable cellular pattern; compare the no-primary control and review blocking, washing and detection (general IHC practice). |
| No signal in a correctly identified nasopharyngeal respiratory epithelium or testicular Leydig-cell control (HPA: High in both). | A negative result is inconclusive until tissue integrity, retrieval, antibody dilution and detection are checked (general IHC practice); HPA levels do not guarantee every section will stain. |
💡Expected ADAM23 appearanceCall an IHC section positive when the relevant cells show distinct cytoplasmic and plasma-membrane staining, potentially strong in nasopharyngeal respiratory epithelium or Leydig cells (HPA: tissue IHC profile; High in both); uniform background or isolated nuclear-only IHC signal is discordant with that profile (HPA: tissue IHC profile; general IHC practice).
| Cell type and tissue | HPA reports High staining in nasopharyngeal respiratory epithelial cells and Leydig cells, Medium in caudate neurons, and Low in several listed cell types (HPA: tissue IHC); compare like cells rather than whole-section colour. |
| Membrane topology and secretion | The mature chain includes an extracellular region at residues 287–792 and a transmembrane segment at 793–813; a secreted location is also listed (UniProt O75077). HPA cautions that secreted variants can complicate RNA–protein agreement (HPA: reliability note). |
| Processing and isoforms | UniProt lists a signal peptide, propeptide, mature chain and Alpha, Beta and Gamma isoforms (UniProt O75077). Without an antibody epitope in the supplied evidence, these features cannot predict which forms a given IHC stain detects. |
| Antibody validation | CAB033276 is listed as IHC Approved, while HPA012130 has ICC Approved status and no IHC status in the supplied antibody record (HPA: antibody validation). Approval supports the stated application; it does not establish identical staining between antibodies. |
| IF/ICC Q&A: should nuclear staining be expected? | HPA approves nucleoplasm and plasma membrane as ICC-IF locations and lists Rh30 and U2OS images (HPA: subcellular ICC-IF). Assess that IF finding in its own application; the tissue IHC profile reports general cytoplasmic expression and plasma positivity (HPA: tissue IHC). |
| Retrieval and detection | Retrieval, blocking and chromogen controls are routine paraffin-IHC variables (general IHC practice). The supplied UniProt and HPA records do not establish ADAM23-specific fixation sensitivity or an optimal retrieval condition. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a known-positive cell population (HPA: High in nasopharyngeal respiratory epithelium and Leydig cells). | An unsuccessful retrieval, unsuitable dilution or failed detection step can suppress IHC signal (general IHC practice); the supplied sources give no ADAM23-specific retrieval setting. | Verify tissue and cell identity, then check the run control, retrieval record, antibody dilution and detection reagents (general IHC practice). |
| Only nuclear staining appears in tissue IHC (HPA: tissue IHC profile). | The result is discordant with HPA's cytoplasmic and plasma-positive tissue profile; HPA's approved nucleoplasmic location comes from ICC-IF (HPA: tissue IHC; subcellular ICC-IF). | Review controls and compare staining with the expected cell populations; do not transfer the ICC-IF nuclear call to paraffin IHC without application-specific evidence (HPA: antibody validation and location records). |
| Colour appears broadly in unexpected cell types (HPA: tissue IHC cell-level results). | Cross-reactivity or endogenous detection activity can produce misleading chromogenic staining (general IHC practice); the appearance alone cannot identify which cause applies. | Inspect a no-primary control, check endogenous-activity blocking appropriate to the detection chemistry, and reassess cellular morphology (general IHC practice). |
| The whole section has diffuse background (general IHC practice). | Inadequate blocking or washing, or excessive detection signal, can reduce cellular contrast (general IHC practice). | Compare with the no-primary control and review blocking, washes, dilution and chromogen development (general IHC practice). |
| Caudate neurons stain less strongly than a High-staining control (HPA: Medium in caudate neurons; High in the specified nasopharyngeal and testicular cells). | The difference may reflect HPA's reported cell-level intensities; unequal staining alone does not demonstrate assay failure (HPA: tissue IHC). | Score the identified neuronal cells against an appropriate control and avoid requiring High intensity in a population reported as Medium (HPA: tissue IHC; general IHC practice). |
| A Low-staining population looks unstained (HPA: Low in the listed glial, epithelial, glandular, stromal or cholangiocyte populations). | Low reported staining offers a weak benchmark; section conditions and scoring thresholds also affect whether faint colour is called positive (HPA: tissue IHC; general IHC practice). | Use a High-staining cell population to check assay performance, and record the Low population as below the scoring threshold if appropriate (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ADAM23 staining
A07382 · IHCImmunohistochemistry (IHC) analyzes of ADAM23 pAb in paraffin-embedded human rectum carcinoma tissue at 1:50,showing strong cytoplasmic stai
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000114948-ADAM23/tissue/cerebral+cortex#imid_7765608
HPA · Parathyroid glandHuman Protein Atlas IHC, Parathyroid gland. Source: https://www.proteinatlas.org/ENSG00000114948-ADAM23/tissue/parathyroid+gland#imid_7771195
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000114948-ADAM23/tissue/colon#imid_7771238
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000114948-ADAM23/tissue/liver#imid_7771231
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA012130; green = ADAM23, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA012130; green = ADAM23, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ADAM23 IHC & IF
🧪Run nasopharynx first and score staining in respiratory epithelial cells (HPA: High in respiratory epithelial cells). HPA detects ADAM23 in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; cells outside the stained epithelium can provide an internal background reference if they remain unstained, but should not be assumed ADAM23-negative (HPA: no negative tissue rows).
Positive control tissue: Nasopharynx (Respiratory epithelial cells, HPA High)
Negative control tissue: None in HPA: ADAM23 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ADAM23 in Rh30, U2OS, with annotated localisation: Nucleoplasm (approved), Plasma membrane (approved) (HPA subcellular).
Technical controls: Include a secondary-only slide with primary antibody omitted, a concentration-matched rabbit polyclonal IgG isotype control, and ADAM23 knockout material or a validated peptide-block control if available (selected A07382 caption: rabbit pAb and PBS in place of primary; standard IHC specificity controls). For chromogenic nasopharynx IHC, block endogenous peroxidase and check for signal from any inflammatory cells on the no-primary slide (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the selected A07382 paraffin-section caption does not report a fixative (selected A07382 caption: fixative not stated). The evidence does not establish whether frozen sections or IF/ICC are easier for ADAM23, although ICC-IF images exist for Rh30 and U2OS (HPA: ICC-IF images in Rh30 and U2OS). In nasopharynx, endogenous peroxidase in inflammatory cells can complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for ADAM23
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex.). 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 |
|---|---|---|---|---|
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: ADAM23 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced ADAM23 IHC Tips
Troubleshoot ADAM23 chromogenic IHC in paraffin sections using the stated retrieval setting, the selected antibody image, and compartment aware controls.
What retrieval should I try first when ADAM23 staining is weak?
Start with citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 minutes (page retrieval setting). After cooling, compare an ADAM23 positive control with a primary antibody omission control on the same staining run, and keep section thickness and detection conditions consistent (standard IHC practice). The extracellular region spans residues 287–792, while the cytoplasmic tail spans 814–832; retrieval performance may therefore depend on which epitope the antibody recognizes (UniProt O75077 topology). If staining remains weak, test a more vigorous retrieval condition on adjacent sections and check whether background rises before adopting it (standard IHC practice).
Could fixation explain weak or uneven ADAM23 staining?
ADAM23 specific sensitivity to fixation is unknown from the supplied evidence, and the selected tissue image reports paraffin embedding without naming a fixative (A07382 tissue IHC caption). Record the fixative, time in fixative, and processing history for every section before comparing staining intensity across specimens (standard IHC practice). On matched sections, hold citrate retrieval at pH 6.0, 95–98 °C, and 20 minutes constant while comparing tissue preservation and staining (page retrieval setting; standard IHC practice). Do not assign a fixation effect from ADAM23 glycosylation or topology alone; interpret any observed difference with matched controls and documented processing (UniProt O75077 record; standard IHC practice).
Should cytoplasmic ADAM23 staining count as a positive IHC result?
Yes, cytoplasmic signal can be compatible with the supplied tissue evidence: the selected paraffin section shows strong cytoplasmic staining, and HPA describes general cytoplasmic expression with plasma positivity (A07382 tissue IHC caption; HPA tissue IHC). Also examine membrane associated signal, because ADAM23 has an extracellular region at residues 287–792 and a transmembrane segment at 793–813 (UniProt O75077 topology). Score cytoplasmic and membrane patterns separately on intact cells, using the primary antibody omission control to judge nonspecific chromogen deposition (standard IHC practice; A07382 tissue IHC caption). Treat an isolated nuclear pattern cautiously in tissue IHC, although HPA reports approved nucleoplasmic and plasma membrane localisation in its subcellular dataset (HPA subcellular).
How can epitope placement change ADAM23 IHC interpretation?
First establish the catalog antibody's immunogen or mapped epitope before treating different staining patterns as biological differences (standard IHC practice). ADAM23 has three reported isoforms, Alpha, Beta, and Gamma, and its signal peptide, propeptide, and mature chain occupy residues 1–59, 60–286, and 287–832, respectively (UniProt O75077 record). An antibody recognizing an extracellular epitope may detect membrane associated or released material, whereas a cytoplasmic tail epitope asks a different localisation question (UniProt O75077 topology; standard IHC interpretation). If epitope information is unavailable, report the observed compartment and antibody identity without claiming isoform specificity or a particular processing state (standard IHC practice).
How should I investigate an ADAM23 pattern with IF alongside IHC?
Use IF as a separate localisation check alongside the chromogenic IHC result, because HPA reports approved nucleoplasmic and plasma membrane localisation in its subcellular dataset (HPA subcellular). For multiplexing, pair ADAM23 with a marker for the cell population under study; HPA reports ADAM23 staining in caudate neuronal cells and nasopharyngeal respiratory epithelial cells (HPA tissue IHC; standard IF practice). Choose a fluorophore in a low autofluorescence channel for the specimen, and include single stain and primary antibody omission controls before interpreting overlap (standard IF practice). Match permeabilisation to the epitope: an extracellular epitope can be examined without membrane permeabilisation, while a cytoplasmic tail epitope requires access to the cell interior (UniProt O75077 topology; standard IF practice).
What controls help separate ADAM23 signal from chromogenic background?
Compare the stained section with a matched primary antibody omission section; the selected image used PBS in place of primary antibody as its negative control (A07382 tissue IHC caption). For a peroxidase chromogen workflow, block endogenous peroxidase before detection and inspect tissue edges, damaged areas, and pigment for deposits that recur in the negative control (standard IHC practice). The selected image used a biotin secondary followed by avidin peroxidase, so include a detection reagent control if diffuse background persists (A07382 tissue IHC caption; standard IHC practice). Reduce primary antibody concentration or detection exposure in a controlled titration, comparing each condition with the reported 1:50 image condition (A07382 tissue IHC caption; standard IHC practice).
How should I quantify ADAM23 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because the supplied observations include cytoplasmic staining and plasma positivity (A07382 tissue IHC caption; HPA tissue IHC). Within a fixed tissue area, record the percentage of positive cells and an intensity based H-score separately for cytoplasmic and membrane signal, or report positive cell density per mm² when cell distribution is the main outcome (standard IHC practice). Normalize counts to the number of evaluable cells or the measured viable tissue area, and exclude folds, necrosis, and detached edges consistently (standard IHC practice). Use the same retrieval, chromogen development, imaging settings, and scoring thresholds across compared sections (page retrieval setting; standard IHC practice).
When is an apparent ADAM23 positive result likely to be an artefact?
Favor a result that follows intact cell boundaries or plausible cytoplasmic distribution and clears in the primary antibody omission control; those patterns align with the supplied localisation and image evidence (UniProt O75077 topology; HPA tissue IHC; A07382 tissue IHC caption). Be cautious when signal concentrates only at cut edges, tissue folds, or necrotic areas, or when the same deposit appears without primary antibody (standard IHC practice). Apparent nuclear only staining needs separate scrutiny in tissue IHC, despite approved nucleoplasmic localisation in HPA subcellular data (HPA subcellular; standard IHC practice). Check endogenous peroxidase and, where relevant, detection reagent background before interpreting DAB deposits as ADAM23 expression in a particular cell population (standard IHC practice).
Boster reagents
Best ADAM23 / Disintegrin and metalloproteinase domain-containing protein 23 IHC Antibodies
A07382 has real IHC data from paraffin-embedded human rectum carcinoma; the catalog also lists mouse reactivity (A07382 image caption; catalog: reactivity).
Real IHC data
Anti-ADAM23 Antibody
Cat # A07382
A07382 is listed for IHC and shows cytoplasmic staining in paraffin-embedded human rectum carcinoma at 1:50 (catalog: applications; A07382 image caption). Mouse reactivity is listed, but no mouse IHC or IF/ICC image is supplied (catalog: reactivity and image fields).
Which to pick: Choose A07382 for tissue IHC: it is a rabbit polyclonal antibody listed for IHC, with an image from paraffin-embedded human rectum carcinoma (catalog: host and applications; A07382 image caption). The caption does not report the fixative (A07382 image caption). A07382 also lists mouse reactivity, but its image shows human tissue; there is no IF/ICC option in this payload (catalog: reactivity, applications and IF fields; A07382 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75077 (ADA23_HUMAN, Disintegrin and metalloproteinase domain-containing protein 23).
- Human Protein Atlas. ADAM23 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ADAM23 subcellular location (ICC-IF): Localized to the nucleoplasm and plasma membrane..
- Human Protein Atlas. ADAM23 antibody validation summary (2 antibodies).
- Identification of ADAM23 as a Potential Signature for Psoriasis Using Integrative Machine-Learning and Experimental Verification. International journal of general medicine 2023 — PMC10750783.
- A disintegrin and metalloprotease 23 hypermethylation predicts decreased disease-free survival in low-risk breast cancer patients. Cancer science 2019 — PMC6500989.
- Based on Molecular Profiling of Gene Expression, Palmoplantar Pustulosis and Palmoplantar Pustular Psoriasis Are Highly Related Diseases that Appear to Be Distinct from Psoriasis Vulgaris. PloS one 2016 — PMC4859542.
- A novel program of infiltrative control in astrocytomas: ADAM23 depletion promotes cell invasion by activating γ-secretase complex. Neuro-oncology advances 2023 — PMC10681280.
- PubMed PMID:9693107 — UniProt-cited evidence.
- PubMed PMID:10749942 — UniProt-cited evidence.
- PubMed PMID:14697522 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ADAM23 (Disintegrin and metalloproteinase domain-containing protein 23). Expected localisation Cytoplasmic expression and plasma positivity (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07382 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Nasopharynx (Respiratory epithelial cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human rectum carcinoma tissue; fixative not specified (datasheet A07382); 2. Fixation: Image fixative and duration unreported (datasheet A07382); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-ADAM23, 1:50-1:200 (datasheet A07382); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07382); Caveat: A secreted variant may shift RNA and protein tissue patterns (HPA tissue IHC); Isoform / epitope: 3 isoforms; check extracellular versus cytoplasmic epitope (UniProt); Regulation: Brain enriched; heart weak (UniProt). Evidence: UniProt O75077, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and plasma membrane., Datasheet A07382, PMC10750783, PMC6500989. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ADAM23/immunohistochemistry-ihc-guide.