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RECK / Reversion-inducing cysteine-rich protein with Kazal motifs · IHC design guide
Design Immunohistochemistry for RECK
Plan RECK paraffin IHC around distinct extracellular matrix staining and additional cytoplasmic staining in several tissues (HPA tissue IHC). Breast myoepithelial cells provide a high-staining reference (HPA tissue IHC); the IHC-validated antibody has a starting dilution range of 1:100–1:300 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RECK 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 | ECM and cytoplasm (HPA tissue IHC); membrane (UniProt) | |
| Staining pattern | Distinct ECM; cytoplasm in several tissues; high in breast myoepithelial cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | ECM positivity can complicate cell-level scoring (HPA tissue IHC) | |
| Regulation | Undetectable in transformed cell lines (UniProt) | |
| Isoform / epitope | No isoforms listed; map epitopes to mature chain 23–942 (UniProt) |
Section 1
Recommended RECK IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published chromogenic RECK IHC methods for salivary adenoid cystic carcinoma (PMC3997680) and paired leiomyoma and myometrium (PMC4988044).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A06439) |
| Fixation | Image fixative and duration unreported (datasheet A06439); 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-RECK, 1:100 - 1:300 (datasheet A06439) |
| 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 | RECK-positive staining in myoepithelial cells of breast (HPA tissue IHC: High). HPA tissue profile: Distinct positivity in extracellular matrix with additional cytoplasmic expression in several tissues. 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); a published RECK method also uses citrate pH 6.0 (PMC3997680).
Section 2
What Is the Expected RECK Staining Pattern?
RECK is a cell membrane protein with no transmembrane segment (UniProt O95980 topology). In tissue IHC, expect distinct extracellular matrix positivity and some cytoplasmic staining (HPA: tissue IHC profile; reliability Supported). Breast myoepithelial cells are a strong reference population (HPA: High); several other annotated cell populations show medium staining (HPA: tissue IHC). Interpret signal by its location and the specific cells stained, rather than by whole-tissue positivity.
| Distinct extracellular matrix staining, with signal along cell borders in an annotated positive population. | This fits the reported extracellular matrix pattern (HPA: tissue IHC profile) and cell membrane localization (UniProt O95980). Record which cells and structures carry signal; extracellular staining alone does not identify its cellular source. |
| Strong staining in breast myoepithelial cells, with lighter staining in other annotated positive populations. | This is consistent with breast myoepithelial cells at High and, for example, colon goblet cells and kidney glomerular cells at Medium (HPA: tissue IHC). These are relative HPA observations, not universal intensity thresholds for every section. |
| Dominant nuclear staining without the expected extracellular, border, or reported cytoplasmic pattern. | Nuclear localization is absent from the supplied localization records (UniProt O95980: cell membrane; HPA: tissue IHC profile). Treat a nuclear-only result as suspect and review controls and detection background before assigning it to RECK. |
| Strong signal in an HPA-annotated undetected cell population, such as adipocytes or bronchial respiratory epithelial cells. | Those specific populations are Not detected (HPA: tissue IHC). Check the stained cell identity and controls; cross-reactivity or endogenous detection activity may explain discordance. An undetected cell population does not make its entire tissue a negative control. |
| No convincing signal in breast myoepithelial cells on a section where those cells can be identified. | That conflicts with the High reference observation (HPA: tissue IHC). Review section quality, antibody and detection controls, and whether the intended cells are present before interpreting other negative populations as true absence. |
💡Expected RECK appearanceCall IHC positive when distinct extracellular matrix staining and appropriate cell-associated signal occur in annotated populations, especially High breast myoepithelial cells (HPA: tissue IHC); isolated nuclear staining or strong signal in an annotated undetected cell population is suspect (UniProt O95980 localization; HPA: tissue IHC).
| Compartment and topology | RECK is assigned to the cell membrane and has no transmembrane segment (UniProt O95980 topology); HPA reports extracellular matrix positivity plus some cytoplasmic expression in tissue (HPA: tissue IHC profile). Interpret these reported patterns together instead of requiring every positive cell to show only a crisp membrane rim. |
| Cell-level reference pattern | Breast myoepithelial cells are High; Purkinje cells, cortical neuropil, colon goblet cells, endometrial stromal cells, glomerular cells, cholangiocytes, and alveolar cells are Medium (HPA: tissue IHC). HPA also lists some populations as Low or Not detected; specify the cell population when comparing sections. |
| Antibody validation | The catalog antibody CAB025109 has Supported IHC status (HPA: antibodies). The tissue profile has Supported reliability, meaning external characterization supports staining (HPA: tissue IHC); neither label establishes that every stained structure is annotated or that every preparation yields the same intensity. |
| Processing and epitope interpretation | UniProt annotates a signal peptide at residues 1–22 and a mature chain at 23–942 (UniProt O95980 processing). Without an epitope position in the supplied record, these annotations cannot predict whether a particular antibody will detect each processed form or explain a missing IHC signal. |
| IF/ICC Q&A: What localization should I expect? | Plasma membrane localization is supported in ICC-IF (HPA: subcellular). That observation helps interpret localization, but ICC-IF evidence is separate from the tissue IHC pattern and does not supply an IF/ICC protocol or establish identical staining intensity in paraffin sections. |
| Situation | Likely cause | Next action |
|---|---|---|
| Breast myoepithelial cells show no signal. | An expected High population is negative (HPA: tissue IHC); the supplied sources do not identify a RECK-specific fixation or retrieval failure. | General IHC practice: confirm the cells are present, check section and detection controls, then review the catalog antibody's IHC-P instructions before changing retrieval or dilution. |
| Staining is predominantly nuclear. | The dominant location conflicts with cell membrane localization and the reported tissue pattern (UniProt O95980; HPA: tissue IHC). Background or cross-reactivity is possible. | General IHC practice: compare with a no-primary control, inspect the counterstain separately, and reassess compartment calls in an annotated positive cell population. |
| Adipocytes or bronchial respiratory epithelial cells stain strongly. | Those cell populations are Not detected in HPA tissue IHC (HPA: tissue IHC); unexpected signal may reflect cell misidentification, cross-reactivity, or endogenous detection activity. | General IHC practice: verify cell morphology and review no-primary and detection-system controls. Do not label the whole adipose or bronchial section negative from one annotated cell type. |
| Nearly every compartment shows diffuse chromogen. | The distribution does not resemble distinct extracellular matrix positivity with limited additional cytoplasmic expression (HPA: tissue IHC profile). Nonspecific binding or detection background is possible. | General IHC practice: examine no-primary and detection controls, review blocking and wash steps, and score only interpretable cell and compartment patterns. |
| Only faint staining appears in a Low HPA population. | HPA lists hippocampal and caudate neuronal cells, among others, as Low (HPA: tissue IHC); a faint result there is a weak performance check. | Use an annotated High population, such as breast myoepithelial cells (HPA: tissue IHC), to judge whether the IHC run detected an expected signal. |
| An ICC-IF image looks more membrane-restricted than the tissue IHC section. | ICC-IF supports plasma membrane localization (HPA: subcellular), while tissue IHC reports extracellular matrix positivity and some cytoplasmic expression (HPA: tissue IHC profile). | Interpret each application against its own reported pattern. For this paraffin-section result, assess tissue cell identity and extracellular staining; consult the separate IF/ICC guide for that application. |
Experiment evidence
IHC & IF images showing RECK staining
A06439 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using RECK Antibody. The picture on the right is blocked with the syn
A06439 · IFImmunofluorescence analysis of HepG2 cells, using RECK Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000122707-RECK/tissue/cerebral+cortex#imid_6688864
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000122707-RECK/tissue/colon#imid_6688790
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000122707-RECK/tissue/liver#imid_6688775
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000122707-RECK/tissue/kidney#imid_6688792
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA071729; green = RECK, blue = nucleus, red = microtubules.
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA071729; green = RECK, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RECK IHC & IF
🧪Run breast first and expect staining in myoepithelial cells (High; HPA: breast myoepithelial cells). Use adipose tissue as a negative comparison, scoring adipocytes, where RECK was not detected (HPA: adipose tissue adipocytes). On the breast slide, compare myoepithelial staining with adjacent non-myoepithelial cells; their RECK status is not specified by the supplied HPA row (HPA: breast myoepithelial cells).
Positive control tissue: Breast (Myoepithelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RECK in ASC52telo, BJ [Human fibroblast], with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a control matched to the primary antibody’s host species and clonality; use nonimmune IgG for a polyclonal primary or an irrelevant antibody of the same isotype for a monoclonal primary (standard IHC practice). A synthesized-peptide block is a documented specificity control for the catalog antibody (A06439 tissue-IHC caption); for chromogenic breast sections, quench endogenous peroxidase and assess background before scoring (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and its effect on RECK staining are unreported, and the selected paraffin-section caption does not state the fixative (A06439 tissue-IHC caption). Retrieval dependency and whether frozen sections or IF are easier are unreported; available ICC-IF images support plasma-membrane localisation but do not establish either comparison (HPA subcellular: plasma membrane; ASC52telo and BJ images). In breast sections, gland boundaries can complicate assignment of chromogenic signal to myoepithelial cells, so score against tissue morphology and the control slides (HPA: breast myoepithelial cells; standard IHC practice).
HPA tissue IHC evidence for RECK
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — External characterization data supports antibody staining. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Breast | Myoepithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Colon | Goblet cells | Medium | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| 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 → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RECK IHC Tips
Troubleshoot RECK staining in paraffin-section chromogenic IHC using the page retrieval conditions, compartment-aware controls, and separately interpreted IF evidence.
How should I retrieve RECK in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page antigen-retrieval rule). Allow sections to cool in the retrieval buffer, then run the same antibody dilution and chromogenic detection across a small retrieval-time series so changes can be attributed to retrieval (standard IHC practice). If staining remains weak, test an alternative buffer or pH as a fallback on adjacent sections, while checking whether tissue structure and the expected staining pattern are preserved (standard IHC practice; HPA: tissue IHC profile). Record the chosen condition before comparing specimens quantitatively (standard IHC practice).
Could fixation explain variable RECK staining between paraffin blocks?
RECK-specific sensitivity to fixation is unknown from the supplied evidence; the catalog antibody’s brain image identifies paraffin embedding but does not state a fixative (A06439 tissue-IHC caption). Compare blocks with documented fixation and processing histories, and keep section thickness, retrieval, antibody incubation, and chromogenic development consistent while investigating variation (standard IHC practice). A stronger signal after changing retrieval does not by itself establish that fixation caused the original difference, so compare adjacent sections and include a known staining control in each run (standard IHC practice). Do not assign a RECK-specific fixation effect from its membrane location or glycosylation (UniProt O95980 topology and glycosylation).
Should RECK appear at the cell surface or in extracellular regions?
Prioritize a membrane-associated pattern when assessing cellular RECK because UniProt places it at the cell membrane and HPA supports plasma-membrane localization (UniProt O95980 subcellular location; HPA subcellular: plasma membrane). In tissue IHC, HPA also reports distinct extracellular-matrix positivity and additional cytoplasmic expression, so annotate those compartments separately instead of forcing every signal into a membrane score (HPA tissue IHC profile). Compare staining with tissue architecture and a no-primary control before accepting diffuse extracellular color as specific (standard IHC practice). The absence of a transmembrane segment does not require purely cytoplasmic staining; the record includes a GPI-anchor annotation (UniProt O95980 topology and keywords).
How can epitope location affect interpretation of RECK IHC?
Check the catalog antibody’s stated immunogen or epitope before interpreting a lost signal, because the supplied record gives no antibody epitope coordinates (supplied A06439 caption; UniProt O95980 record). RECK has a signal peptide at residues 1–22, a mature chain at 23–942, and a propeptide at 943–971 (UniProt O95980 processing). Five listed glycosylation sites and three Kazal-like domains make epitope accessibility a reasonable experimental concern, but they do not identify this antibody’s binding site (UniProt O95980 glycosylation and domains). The record lists 0 isoforms, so do not explain differing section patterns by asserting an unreported isoform (UniProt O95980 isoforms).
How should I investigate RECK localization with multiplex IF alongside IHC?
Use IF as a separate localization check and pair RECK with a validated marker for the expected cell population, such as a marker selected for breast myoepithelial cells when studying that population (HPA tissue IHC: high in breast myoepithelial cells; standard IF practice). Choose fluorophores after measuring autofluorescence in unstained sections, and place the weaker channel where tissue background is lowest (standard IF practice). Set permeabilisation according to the verified antibody epitope: minimize it for an accessible surface epitope, or optimize it for an intracellular epitope (standard IF practice). The supplied record establishes plasma-membrane localization but does not map this antibody’s epitope (HPA subcellular; supplied A06439 caption).
What controls help separate RECK signal from chromogenic background?
Include no-primary and appropriate secondary-only controls to reveal nonspecific detection, and compare them with sections stained using the IHC-validated antibody (standard chromogenic IHC practice). Block endogenous peroxidase before an enzyme-based DAB reaction and adjust development time if diffuse brown deposit obscures cell boundaries (standard chromogenic IHC practice). The catalog image includes a peptide-blocked comparison in paraffin-embedded human brain, which supports checking that image’s staining under its stated conditions but does not establish every tissue pattern (A06439 tissue-IHC caption). Review section edges, folds, pigment, and necrotic regions separately from intact tissue before calling widespread extracellular staining specific (standard IHC practice; HPA tissue IHC: extracellular-matrix positivity).
How should I score RECK across specimens with different tissue composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring: membrane-associated cellular signal and extracellular-matrix signal can coexist in the reported patterns (UniProt O95980 subcellular location; HPA tissue IHC profile). For a cellular endpoint, report an H-score or percent positive cells within a prespecified population, using the number of evaluable cells in that population as the denominator (standard IHC quantification practice). For extracellular signal, quantify positive area or optical density per mm² of evaluable tissue or selected region, excluding folds and necrosis by a consistent rule (standard IHC quantification practice). Keep retrieval, chromogen development, imaging, and threshold settings matched across specimens (standard IHC practice).
When should I question a positive RECK IHC result?
Question staining restricted to an unexpected compartment or cell population, and compare it with the documented membrane localization and tissue-specific IHC observations before assigning it to RECK (UniProt O95980 subcellular location; HPA tissue IHC profile). For example, HPA reports high staining in breast myoepithelial cells and no detection in adipocytes, so score those populations separately when present (HPA tissue IHC: breast and adipose tissue). Edge-only color, necrotic deposits, or signal reproduced in a no-primary or peroxidase-control section favors an artefact requiring protocol review (standard IHC practice). Peptide blocking in the catalog brain image is useful context, but tissue localization still needs appropriate controls and morphology (A06439 tissue-IHC caption; standard IHC practice).
Boster reagents
Best RECK / Reversion-inducing cysteine-rich protein with Kazal motifs IHC Antibodies
Anti-RECK A06439 has IHC data from paraffin-embedded human brain and IF data from HepG2 cells (catalog image captions); listed reactivity is human and mouse (catalog reactivity).
Real IHC data
Anti-RECK Antibody
Cat # A06439
A06439 will render with its IHC image of paraffin-embedded human brain tissue (catalog IHC image caption). The same SKU has an IF image of HepG2 cells and lists IHC, IF and ICC applications with human and mouse reactivity (catalog IF image caption; catalog applications; catalog reactivity).
Which to pick: Choose A06439 for tissue IHC in paraffin sections, as shown in its own human brain image; the fixative is unreported (catalog IHC image caption). Choose A06439 for IF/ICC based on its listed applications and HepG2 IF image (catalog applications; catalog IF image caption). For human or mouse samples, A06439 is the listed rabbit polyclonal option, although the supplied images show human brain tissue and HepG2 cells only (catalog reactivity; catalog host and clonality; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95980 (RECK_HUMAN, Reversion-inducing cysteine-rich protein with Kazal motifs).
- Human Protein Atlas. RECK tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RECK subcellular location (ICC-IF): Localized to the plasma membrane..
- Human Protein Atlas. RECK antibody validation summary (2 antibodies).
- Expression of RECK and MMP-2 in salivary adenoid cystic carcinoma: Correlation with tumor progression and patient prognosis. Oncology letters 2014 — PMC3997680.
- RECK controls breast cancer metastasis by modulating a convergent, STAT3-dependent neoangiogenic switch. Oncogene 2015 — PMC4344427.
- MicroRNA expression in lung tissues of asbestos-exposed mice: Upregulation of miR-21 and downregulation of tumor suppressor genes Pdcd4 and Reck. Journal of occupational health 2021 — PMC8535435.
- MicroRNA-15b regulates reversion-inducing cysteine-rich protein with Kazal motifs (RECK) expression in human uterine leiomyoma. Reproductive biology and endocrinology : RB&E 2016 — PMC4988044.
- PubMed PMID:9789069 — UniProt-cited evidence.
- PubMed PMID:15164053 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RECK (Reversion-inducing cysteine-rich protein with Kazal motifs). Expected localisation ECM and cytoplasm (HPA tissue IHC); membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A06439 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Myoepithelial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A06439); 2. Fixation: Image fixative and duration unreported (datasheet A06439); 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-RECK, 1:100 - 1:300 (datasheet A06439); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: ECM positivity can complicate cell-level scoring (HPA tissue IHC); Isoform / epitope: No isoforms listed; map epitopes to mature chain 23–942 (UniProt); Regulation: Undetectable in transformed cell lines (UniProt). Evidence: UniProt O95980, HPA tissue IHC · Supported, HPA ICC-IF · plasma membrane., Datasheet A06439, PMC3997680, PMC4988044. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RECK/immunohistochemistry-ihc-guide.