ADAM23 / Disintegrin and metalloproteinase domain-containing protein 23 · IHC design guide

Design Immunohistochemistry for ADAM23

For ADAM23 paraffin IHC, nasopharynx respiratory epithelium and testis Leydig cells provide high-staining references (HPA tissue IHC). Start the IHC-validated antibody at 1:50–1:200 (datasheet) and assess cytoplasmic and plasma membrane staining (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for ADAM23 (IHC for ADAM23): expected localisation Cytoplasmic and plasma membrane staining (HPA tissue IHC), antibody A07382, validated IHC image, and IHC protocol steps — Printable **ADAM23** IHC protocol sheet — expected localisation Cytoplasmic and plasma membrane staining (HPA tissue IHC), antibody A07382, controls and protocol steps. Open the full ADAM23 IHC guide →

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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 and plasma membrane staining (HPA tissue IHC)
	Staining pattern	Cytoplasmic/plasma signal in respiratory epithelial and Leydig cells (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	Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific)
	Caveat	A secreted variant may alter tissue staining (HPA tissue IHC)
	Regulation	No specific regulator reported (UniProt)
	Isoform / epitope	3 isoforms; extracellular vs cytoplasmic epitope coverage unknown (UniProt)

Section 1

Recommended ADAM23 IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol (datasheet) with two published ADAM23 IHC protocols (PMC10750783; PMC6500989).

Recommended immunohistochemistry (IHC-P) protocol parameters
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 retrieval rule); the skin study used 121 °C for 10 min (PMC10750783).

Section 2

What Is the Expected ADAM23 Staining Pattern?

Expect ADAM23 staining at the plasma membrane and in the cytoplasm, with the strongest reported tissue IHC signal in nasopharyngeal respiratory epithelial cells and testicular Leydig cells (HPA: general cytoplasmic expression and plasma positivity; HPA: High in both cell types). Its extracellular region spans residues 287–792 and its membrane segment spans 793–813 (UniProt O75077 topology). HPA rates its tissue IHC profile Approved and cautions that a secreted variant can complicate RNA–protein comparisons (HPA: tissue IHC reliability).

What am I looking at on my slide?
Distinct membrane staining with associated cytoplasmic signal in nasopharyngeal respiratory epithelium or Leydig cells.	This fits the reported compartments and High staining in those cell types (HPA: tissue IHC profile; HPA: High in respiratory epithelial and Leydig cells). Judge signal against nearby cells and the negative control under matched detection conditions (general IHC practice).
Predominantly nuclear chromogenic staining, without a convincing membrane or cytoplasmic pattern.	Treat this as discordant with the reported tissue IHC profile and check specificity and counterstain interpretation (HPA: general cytoplasmic expression and plasma positivity; general IHC practice). Nucleoplasmic localization is approved in ICC-IF, but that observation does not by itself validate nuclear staining in paraffin tissue (HPA: ICC-IF subcellular).
Strong staining mainly in cells outside the documented positive cell populations.	Investigate cross-reactivity or endogenous chromogenic activity before calling those cells ADAM23 positive (HPA: tissue IHC cell-level observations; general IHC practice). The HPA list samples selected cells and tissues, so an unlisted cell type is not automatically negative (HPA: tissue IHC).
Uniform color across cells, stroma, and tissue-free areas, with little cell-boundary definition.	This favors background over the reported cytoplasmic and plasma pattern (HPA: tissue IHC profile; general IHC practice). Check the matched negative control, blocking, washes, and detection exposure; those are general workflow checks, not documented ADAM23-specific effects (general IHC practice).
No detectable staining in nasopharyngeal respiratory epithelium or testicular Leydig cells.	A negative run in either reported High cell population is inconclusive until slide quality, antibody conditions, and detection controls are checked (HPA: High in both cell types; general IHC practice). HPA's Approved profile does not guarantee positivity with every antibody or specimen (HPA: tissue IHC reliability).

💡Expected ADAM23 appearanceA credible positive shows membrane and cytoplasmic staining in the documented cell populations, especially High signal in nasopharyngeal respiratory epithelial or testicular Leydig cells; diffuse field-wide color or nuclear-only tissue staining needs control-based review (HPA: tissue IHC profile and levels; general IHC practice).

How each factor affects the staining
Cell type and tissue	HPA reports High staining in nasopharyngeal respiratory epithelium and Leydig cells, Medium staining in caudate neuronal cells, and Low staining in hippocampal glial cells (HPA: tissue IHC). Score the identified cell population rather than the whole section (general IHC practice).
Membrane topology and secreted form	ADAM23 has an extracellular region, one transmembrane segment, and a short cytoplasmic tail; UniProt also lists it as secreted (UniProt O75077 topology and subcellular). HPA warns that at least one variant is secreted, so extracellular signal warrants careful morphological and control review (HPA: reliability description; general IHC practice).
Processing, isoforms, and epitope	UniProt lists a signal peptide, propeptide, mature chain, and three isoforms (UniProt O75077 processing and isoforms). The supplied record gives no antibody epitope or isoform coverage; do not infer which forms an observed stain detects.
Antibody validation	One listed rabbit antibody has IHC Approved status, while a different rabbit antibody has ICC Approved status (HPA: CAB033276 IHC Approved; HPA: HPA012130 ICC Approved). These statuses are assay-specific and do not establish identical staining for the two antibodies (HPA: antibody validation).
IF/ICC Q: should nuclei stain?	HPA reports approved nucleoplasm and plasma membrane localization in ICC-IF, with images from Rh30 and U2OS (HPA: ICC-IF subcellular). Assess that finding within an IF/ICC experiment; it does not change the tissue IHC expectation of cytoplasmic and plasma positivity (HPA: tissue IHC profile).
Antigen retrieval	Retrieval is a general paraffin IHC variable to optimize with positive and negative controls (general IHC practice). Neither supplied source identifies an ADAM23-specific retrieval condition or fixation sensitivity; an observed change cannot be predicted from its topology or HPA staining levels.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected positive cells have no chromogenic signal.	Possible assay failure, unsuitable antibody conditions, or an uninformative specimen (general IHC practice); HPA reports High signal in nasopharyngeal respiratory epithelial and Leydig cells (HPA: tissue IHC).	Verify tissue identity and morphology, then run the antibody with matched detection and a suitable positive control (general IHC practice). Optimize dilution and retrieval empirically; no target-specific settings are supplied.
Color appears throughout the section, including outside recognizable cells.	Diffuse chromogen may reflect nonspecific binding, endogenous detection activity, or incomplete washing (general IHC practice). It does not resemble HPA's cell-associated cytoplasmic and plasma profile (HPA: tissue IHC).	Compare with reagent-omission and detection controls; review blocking, washing, and chromogen exposure under matched conditions (general IHC practice).
Nuclei dominate the paraffin-section stain.	This differs from HPA's general tissue IHC pattern; approved nucleoplasmic staining comes from ICC-IF observations (HPA: tissue IHC profile; HPA: ICC-IF subcellular).	Check the counterstain and negative control, then reassess whether membrane or cytoplasmic signal is present in documented positive cells (general IHC practice; HPA: tissue IHC).
A presumed negative cell population stains strongly.	Cross-reactivity or endogenous chromogenic activity is possible (general IHC practice). HPA's listed Low cells include hippocampal glia, but Low is not equivalent to absent staining (HPA: tissue IHC).	Confirm cell identity and compare staining with controls and an independently validated IHC antibody if available (general IHC practice). Avoid declaring an unlisted tissue negative from this record.
Signal is extracellular or does not track clear cell boundaries.	UniProt lists secreted and membrane localization, and HPA notes a secreted variant; neither source defines a diagnostic extracellular IHC pattern (UniProt O75077 subcellular; HPA: reliability description).	Inspect morphology and negative controls before assigning extracellular color to ADAM23; record membrane, cytoplasmic, and extracellular signal separately (general IHC practice).
IHC and IF/ICC show different compartments.	HPA reports cytoplasmic and plasma positivity in tissue IHC, and nucleoplasm plus plasma membrane in ICC-IF; the listed assay approvals belong to different antibodies (HPA: tissue IHC, ICC-IF, antibodies).	Interpret each assay with its own validated antibody and controls; do not transfer the ICC-IF nuclear finding into a paraffin IHC scoring rule (HPA: antibody validation and subcellular observations; general IHC practice).

Source: UniProt O75077 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing ADAM23 staining

Immunohistochemistry (IHC) analyzes of ADAM23 pAb in paraffin-embedded human rectum carcinoma tissue at 1:50,showing strong cytoplasmic staining.Negative control (the right)Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG-biotin followed by avidin-peroxidase.

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.

← 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 require 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; cells outside the stained epithelial compartment should show only background, but are not validated ADAM23-negative cells (HPA: no negative tissue listed).

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 no-primary, secondary-only slide, a matched nonimmune rabbit IgG control for the rabbit polyclonal catalog antibody, and, if available, ADAM23-knockout material or a validated immunizing-peptide block (selected tissue-IHC caption: pAb with anti-rabbit secondary). Quench endogenous peroxidase and check for endogenous biotin if using the caption’s avidin–peroxidase detection; inspect nasopharyngeal mucus for background staining (selected tissue-IHC caption: avidin–peroxidase; standard IHC practice).

⚠️Feasibility: Paraffin-section IHC is documented at 1:50, but the selected-SKU caption does not report a fixative, and no target-specific fixation window or retrieval dependency is reported (selected tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier than paraffin IHC (HPA: ICC-IF images in Rh30 and U2OS; selected tissue-IHC caption: paraffin section). In nasopharynx, assess mucus-associated background when scoring respiratory epithelial staining (HPA: High in respiratory epithelial cells; 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 staining in paraffin section IHC by checking retrieval, compartment, controls and scoring before comparing results across tissues.

How should I retrieve ADAM23 antigen when staining is weak?

Start with citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 min (page retrieval rule: cytoplasmic / membrane antigen). Cool sections consistently before staining, and change one retrieval variable at a time while retaining an identically processed control section (standard IHC practice). Compare membrane and cytoplasmic signal separately: ADAM23 has an extracellular region at residues 287–792 and a transmembrane segment at 793–813 (UniProt O75077 topology). If signal remains weak, trial a different retrieval buffer or pH on paired sections and assess background alongside signal (standard IHC practice).

Could fixation account for variable ADAM23 staining between paraffin blocks?

Target specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin section caption does not state its fixative (A07382 tissue IHC caption). Record each block’s fixative and fixation duration, then compare sections processed with the same retrieval and detection conditions (standard IHC practice). Include a control section from a block with established staining in each run to help distinguish a block effect from a run effect (standard IHC practice). Do not assign a fixation cause solely from cytoplasmic or membrane staining: both locations occur in the supplied ADAM23 evidence (HPA tissue IHC profile; UniProt O75077 subcellular annotation).

Where should ADAM23 signal appear in tissue sections?

Assess cytoplasmic and plasma membrane staining as distinct patterns because tissue IHC reports general cytoplasmic expression and plasma positivity (HPA tissue IHC profile). ADAM23 is annotated at the cell membrane and as secreted, with an extracellular region spanning residues 287–792 (UniProt O75077 subcellular annotation and topology). The selected paraffin section image shows strong cytoplasmic staining in human rectum carcinoma at 1:50 (A07382 tissue IHC caption). Record any nuclear signal separately: nucleoplasm and plasma membrane are approved subcellular locations in cell imaging, but that observation alone does not establish the source of nuclear staining in a tissue section (HPA subcellular; standard IHC interpretation).

How can epitope position complicate ADAM23 IHC interpretation?

Confirm the antibody’s stated immunogen or mapped epitope before interpreting a missing compartment or comparing antibodies (standard IHC practice). ADAM23 has 3 annotated isoforms—Alpha, Beta and Gamma—and a signal peptide, propeptide and mature chain spanning residues 287–832 (UniProt O75077 isoforms and processing). An epitope in the propeptide could report a different processed species from one in the mature extracellular region; this is a sequence based possibility, not a demonstrated staining difference (UniProt O75077 processing and topology). Document whether the epitope is known, and avoid calling a negative section isoform negative without isoform specific validation (standard IHC interpretation).

How should I investigate ADAM23 localisation with complementary IF?

Use IF as a complementary localisation experiment, keeping its controls and interpretation separate from the paraffin section IHC result (standard IF practice). Multiplex ADAM23 with a validated marker for the cell type being examined; neuronal cells in caudate have reported medium tissue IHC staining (HPA tissue IHC: caudate neuronal cells). Choose spectrally separated fluorophores and a channel with low measured tissue autofluorescence, using unstained and single stain controls (standard IF practice). Match permeabilisation to the antibody epitope: an extracellular epitope may be accessible without it, whereas a cytoplasmic epitope requires access across the membrane (UniProt O75077 topology; standard IF practice).

What controls help resolve diffuse or patchy ADAM23 chromogenic background?

Inspect a no primary control alongside the stained section; the selected image used PBS in place of primary antibody and a biotin, avidin and peroxidase detection sequence (A07382 tissue IHC caption). Block endogenous peroxidase before chromogen development, and check whether background persists in the no primary section (standard chromogenic IHC practice). With avidin and biotin detection, assess endogenous biotin as another possible source of signal (A07382 tissue IHC caption; standard IHC practice). Compare staining away from section edges and damaged regions, then adjust blocking, washes or antibody concentration one variable at a time (standard IHC practice).

How should I quantify ADAM23 staining across IHC samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, since ADAM23 tissue IHC includes cytoplasmic expression and plasma positivity (HPA tissue IHC profile). For cellular staining, report percent positive cells and an intensity based H-score using the same thresholds across sections (standard IHC practice). For a spatial readout, report positive cell density per mm² of evaluable tissue and specify the sampled regions (standard IHC practice). Normalise counts to evaluable cells or tissue area, and exclude folds, necrosis and section edges using prespecified rules (standard IHC practice). Report membrane and cytoplasmic scores separately so a shift in pattern is visible (HPA tissue IHC profile; standard IHC practice).

How can I distinguish plausible ADAM23 staining from artefact?

Judge signal against the expected cell and compartment patterns, including reported high staining in nasopharyngeal respiratory epithelial cells and testicular Leydig cells (HPA tissue IHC). Cytoplasmic or membrane signal is plausible in tissue, while nuclear staining deserves separate review because nucleoplasm is reported in cell imaging (HPA tissue IHC profile; HPA subcellular). Check whether colour concentrates at section edges, within necrotic areas or in a no primary control before assigning it to ADAM23 (standard IHC practice). Residual signal in the no primary control can indicate detection background, including endogenous enzyme activity in chromogenic IHC (standard IHC practice). Interpret tissue and RNA differences cautiously because at least one ADAM23 variant is secreted (HPA tissue IHC reliability description).

Boster reagents

Best ADAM23 / Disintegrin and metalloproteinase domain-containing protein 23 IHC Antibodies

A07382 has real IHC image data from paraffin-embedded human rectum carcinoma tissue (A07382 image caption); its listed reactivity covers human and mouse (catalog reactivity).

Real IHC data Immunohistochemistry (IHC) analyzes of ADAM23 pAb in paraffin-embedded human rectum carcinoma tissue at 1:50,showing strong cytoplasmic staining.Negative control (the right)Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG-biotin followed by avidin-peroxidase.

Anti-ADAM23 Antibody

Cat # A07382

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A07382 is listed for IHC with human and mouse reactivity (catalog applications/reactivity). Its IHC image shows strong cytoplasmic staining at 1:50 in paraffin-embedded human rectum carcinoma tissue, alongside a primary-antibody omission control (A07382 image caption).

Which to pick: For paraffin-section IHC, choose A07382: it is a rabbit polyclonal antibody (catalog host; A07382 image caption) with a listed IHC dilution of 1:50–1:200 (catalog dilution) and a paraffin-section image; the fixative is unreported (A07382 image caption). No IF/ICC choice is supported because A07382 has no listed IF/ICC application or IF image (catalog applications/images). For human and mouse work, A07382 lists reactivity with both species (catalog reactivity), while its pictured IHC evidence is human only (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.