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SCGB2A1 / Mammaglobin-B · IHC design guide
Design Immunohistochemistry for SCGB2A1
Plan chromogenic IHC for SCGB2A1 in paraffin sections using cytoplasmic gland staining as a reference (HPA tissue IHC). This guide covers fixation, controls, and interpretation when tissue RNA and protein locations differ for a secreted protein (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SCGB2A1 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in uterine and epididymal glands (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cervix+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 consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | RNA and protein locations may differ for this secreted protein (HPA tissue IHC) | |
| Regulation | May be regulated by steroid hormones (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 19–95 (UniProt) |
Section 1
Recommended SCGB2A1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is accompanied by a published gastric cancer tissue microarray protocol (PMC6995300).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A07927) |
| Fixation | Image fixative and duration unreported (datasheet A07927); 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-SCGB2A1, 1:50-1:200 (datasheet A07927) |
| 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 | SCGB2A1-positive staining in glandular cells of cervix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in uterine and epididymal glands. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval); the published protocol used microwave heating for 15 min (PMC6995300).
Section 2
What Is the Expected SCGB2A1 Staining Pattern?
SCGB2A1 is secreted and has no transmembrane segment (UniProt O75556: subcellular location and topology). In paraffin-section IHC, expect predominantly cytoplasmic staining in uterine and epididymal glands (HPA: tissue IHC profile). HPA rates the tissue pattern Enhanced for agreement with RNA data, while noting that secretion can separate protein location from RNA location and that external verification is pending (HPA: reliability description).
| Cytoplasmic staining in uterine or epididymal glandular epithelium, with relatively little staining in surrounding cells. | This matches the reported tissue pattern (HPA: cytoplasmic expression mainly in uterine and epididymal glands). Compare cells within the same section; secretion means extracellular staining may also need context (UniProt O75556: secreted). |
| Strong, sharply nuclear or membrane-restricted staining, with little cytoplasmic signal in the expected glands. | That distribution does not match the reported IHC profile (HPA: cytoplasmic glandular expression) or a membrane-spanning topology (UniProt O75556: no transmembrane segment). Review controls and staining conditions before calling it specific (general IHC practice). |
| Prominent staining in cells outside the reported positive populations, especially when expected glands are unstained. | Treat this as uncertain specificity; cross-reactivity or endogenous detection activity can mimic a positive result (general IHC practice). HPA reports selected cell populations and staining levels, not proof that every other cell must be negative (HPA: tissue IHC). |
| A broad haze covers glands, stroma, and blank areas with little distinction between cells. | This is difficult to score as localized antigen staining (general IHC practice). Compare a no-primary control and assess blocking, washing, and chromogen development before interpreting the glandular signal (general IHC practice). |
| No convincing signal in an adequately preserved cervix, endometrium, or epididymis section. | HPA reports high staining in cervix glandular cells, endometrial ciliated epithelial cells, and epididymal glandular cells (HPA: tissue IHC). Confirm the relevant cells are present, then check the IHC workflow; a negative run alone does not establish absent SCGB2A1 (general IHC practice). |
💡Expected SCGB2A1 appearanceCall a positive when appropriate glandular or ciliated epithelial cells show distinct cytoplasmic staining, potentially high in cervix, endometrium, or epididymis (HPA: tissue IHC); widespread haze or isolated nuclear staining warrants investigation (general IHC practice).
| Tissue and cell selection | Cervix glandular cells, endometrial ciliated epithelial cells, and epididymal glandular cells are reported high; nasopharyngeal ciliated cells and salivary glandular cells are medium (HPA: tissue IHC). Evaluate the named cells, since an entire section has no single expected intensity (general IHC practice). |
| Secreted protein and topology | SCGB2A1 is secreted and lacks a transmembrane segment (UniProt O75556: location and topology). The HPA cytoplasmic IHC pattern describes observed cells; extracellular material should be interpreted cautiously because protein and RNA locations can differ (HPA: reliability description). |
| Protein processing | UniProt annotates a signal peptide at residues 1–18 and a mature chain at 19–95 (UniProt O75556: processing). The supplied evidence does not locate the catalog antibody epitope, so it cannot predict which form the stain detects. |
| Antibody evidence | The listed rabbit polyclonal antibody HPA034584 has Enhanced IHC status (HPA: antibody validation). This supports the reported pattern but does not certify every positive cell or replace tissue and detection controls (general IHC practice). |
| IF/ICC question: should vesicular signal appear in IHC? | Vesicles are the approved ICC-IF location (HPA: subcellular). That observation may fit secretory trafficking (UniProt O75556: secreted), but it does not define a required vesicular pattern in paraffin-section IHC; judge IHC against the tissue profile (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive tissue has no staining. | The sampled section may lack the reported positive cells, or the staining run may have failed (HPA: tissue IHC; general IHC practice). | Check for cervix or epididymis glandular cells or endometrial ciliated epithelial cells (HPA: tissue IHC). Review antibody dilution, retrieval, detection reagents, and run controls using standard IHC practice; no SCGB2A1-specific retrieval condition is supplied. |
| The whole section shows brown haze. | Excess background from detection chemistry, incomplete blocking, or insufficient washing is possible (general IHC practice). | Compare a no-primary control, review blocking and wash steps, and shorten chromogen development if needed (general IHC practice). Score only cellular staining distinguishable from background. |
| Nuclear or sharply membrane-bound signal dominates. | This conflicts with the reported cytoplasmic IHC profile (HPA: tissue IHC) and with the lack of a transmembrane segment (UniProt O75556: topology). | Inspect the counterstain and negative control, then reassess detection background and antibody concentration before assigning SCGB2A1 localization (general IHC practice). |
| Unexpected cells stain more strongly than the expected glands. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA's profile describes particular tissues and cells, not universal absence elsewhere (HPA: tissue IHC). | Compare the unexpected cells with a no-primary control and a positive tissue; record cell type and compartment before scoring (general IHC practice). |
| A low-staining tissue appears negative. | HPA reports low staining in breast glandular cells, fallopian-tube ciliated cells, and several other specified populations (HPA: tissue IHC). | Use an HPA high-staining cell population to assess whether the run worked, then describe the low-staining sample as below detection under the tested conditions (HPA: tissue IHC; general IHC practice). |
| An ICC-IF image looks punctate, but the IHC section looks broadly cytoplasmic. | HPA approves a vesicular ICC-IF location and separately reports cytoplasmic tissue IHC expression (HPA: subcellular; HPA: tissue IHC). | Interpret each image in its assay context; for paraffin IHC, assess the documented cell type and cytoplasmic pattern, with controls for background (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing SCGB2A1 staining
A07927 · IHCImmunohistochemistry (IHC) analyzes of Mammaglobin B (K23) pAb in paraffin-embedded human breast carcinoma tissue.
A07927-1 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue, using Mammaglobin B Antibody. The picture on the right is
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000124939-SCGB2A1/tissue/cerebral+cortex#imid_8115128
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000124939-SCGB2A1/tissue/colon#imid_8113678
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000124939-SCGB2A1/tissue/liver#imid_8113687
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000124939-SCGB2A1/tissue/kidney#imid_8113680
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034584; green = SCGB2A1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA034584; green = SCGB2A1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SCGB2A1 IHC & IF
🧪Run cervix first and expect staining in glandular cells (HPA: High in cervical glandular cells). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the cervical slide, assess non-glandular cells for background staining without assuming they are confirmed SCGB2A1-negative (HPA: cervical glandular cells High).
Positive control tissue: Cervix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SCGB2A1 in A-431, EFO-21, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a host-matched nonimmune IgG control for a polyclonal primary, or a host- and isotype-matched irrelevant antibody for a monoclonal primary (standard IHC practice); add SCGB2A1 knockout material or validated antigen-peptide competition as a biological specificity control (standard IHC practice). Block endogenous peroxidase in cervical sections for chromogenic detection, and check endogenous biotin if using an avidin–biotin system (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and the effect of antigen retrieval are unreported in the supplied evidence; optimize retrieval empirically (standard paraffin-section IHC practice). The selected catalog antibody caption shows staining in paraffin-embedded breast carcinoma but does not report the fixative (caption: A07927); the available IF images show vesicular localization, but do not establish that IF or frozen sections are easier (HPA: vesicles, approved). Because SCGB2A1 is secreted, assess luminal or extracellular signal carefully when scoring cellular staining (UniProt O75556: secreted).
HPA tissue IHC evidence for SCGB2A1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Secreted protein, tissue location of RNA and protein is expected to differ. 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 |
|---|---|---|---|---|
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Ciliated epithelial cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Salivary 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 → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SCGB2A1 IHC Tips
Troubleshoot SCGB2A1 staining in paraffin sections by checking retrieval, controls, cellular location and scoring before interpreting chromogenic signal.
Which retrieval conditions should I try first for weak SCGB2A1 staining?
Start with citrate buffer at pH 6.0 for heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval specification). If staining remains weak, compare a longer citrate incubation or an alkaline retrieval buffer on adjacent sections, keeping antibody concentration and detection conditions constant (standard IHC practice). Use a glandular positive-control section alongside the test tissue, since SCGB2A1 staining is high in cervical and epididymal glandular cells (HPA tissue IHC). Check morphology and background before adopting a stronger condition; excessive retrieval can damage sections or increase nonspecific staining (standard IHC practice).
How should I assess whether fixation is causing variable SCGB2A1 staining?
Target-specific fixation sensitivity is unknown from the supplied evidence; the catalog image identifies paraffin-embedded breast carcinoma but does not report its fixative (selected A07927 tissue-IHC caption). Record fixative, fixation duration and tissue thickness for each specimen, then compare sections processed and stained together (standard IHC practice). Include a consistently processed positive-control section and a no-primary control to separate variable antigen detection from chromogen or tissue background (standard IHC practice). If signal differs between otherwise comparable blocks, assess morphology and retrieval response before attributing the difference to SCGB2A1 biology; fixation and processing can alter epitope accessibility in IHC (standard IHC practice).
Where should convincing SCGB2A1 chromogenic staining appear?
Look for cytoplasmic staining in appropriate glandular epithelium, especially uterine and epididymal glands (HPA tissue IHC: cytoplasmic expression mainly in uterine and epididymal glands). SCGB2A1 is secreted, has no transmembrane segment and localizes to vesicles in subcellular imaging, so cytoplasmic puncta or luminal signal can merit review in the context of tissue architecture (UniProt O75556 topology; HPA subcellular: vesicles). Score epithelial cells separately from luminal material, because secreted protein may appear away from its producing cells (UniProt O75556: secreted; HPA tissue IHC: RNA and protein locations may differ). Treat predominantly nuclear or diffuse stromal staining as suspect until supported by controls and reproducible cellular localization (standard IHC practice).
Could processing or glycosylation explain an unexpected staining pattern?
Check the antibody's stated immunogen or epitope against the mature 19–95 chain, because the 1–18 signal peptide is removed during processing (UniProt O75556 processing). One glycosylation site is annotated at residue 68, so an epitope near that site may warrant comparison across retrieval conditions, although the payload gives no measured effect on antibody binding (UniProt O75556 glycosylation). No isoforms are annotated, making an isoform-specific explanation unsupported by this record (UniProt O75556 isoforms). Compare the same control across staining runs before interpreting a change in distribution as altered processing; section handling and detection can also change apparent signal (standard IHC practice).
How can IF help resolve ambiguous SCGB2A1 IHC staining?
Use IF/ICC as a separate localization check and pair SCGB2A1 with a validated marker of the expected glandular or ciliated epithelial cell type (HPA tissue IHC: glandular and ciliated epithelial staining; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with low tissue autofluorescence, using single-label controls to assess bleed-through (standard IF practice). For a putative intravesicular epitope, compare mild permeabilization after fixation with an unpermeabilized condition; SCGB2A1 is secreted and its approved subcellular location is vesicles (UniProt O75556 topology; HPA subcellular: vesicles). Compare the resulting cell-level pattern with chromogenic sections without assuming that IF and IHC signal intensities are directly interchangeable (standard IF/IHC practice).
What should I check when SCGB2A1 staining appears widespread?
Run a no-primary control and inspect endogenous pigment, endogenous peroxidase and nonspecific secondary-reagent signal before assigning diffuse chromogen to SCGB2A1 (standard chromogenic IHC practice). Use a peroxidase block for an HRP/DAB workflow, then adjust blocking, primary-antibody concentration or wash stringency one variable at a time (standard IHC practice). Compare glandular signal with a control expected to lack staining, such as adipocytes in adipose tissue, while recognizing that this is an HPA observation rather than proof of universal absence (HPA tissue IHC: adipocytes not detected). If only section edges, folds or necrotic areas stain, repeat assessment in intact central tissue (standard IHC practice).
How should I quantify heterogeneous SCGB2A1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the epithelial compartment and exclude folds, necrosis and acellular luminal material before scoring, so deposited secreted protein does not inflate cell-based results (UniProt O75556: secreted; standard IHC practice). Record the percentage of positive cells and intensity on a prespecified 0–3 scale, then calculate an H-score from 0–300 if intensity-weighted scoring is needed (standard IHC scoring practice). For spatial analysis, report positive-cell density per mm² of viable epithelial area and retain luminal staining as a separate observation (standard IHC practice). Normalize comparisons to the same defined cell population, tissue area, staining batch and positive-control performance (standard IHC practice).
When is an apparent SCGB2A1 positive result credible?
Favor reproducible cytoplasmic staining in morphologically appropriate glandular cells, with a clean no-primary control and acceptable positive-control staining (HPA tissue IHC: glandular cytoplasmic profile; standard IHC practice). Interpret luminal signal cautiously because SCGB2A1 is secreted, and assess whether nearby epithelial cells show a coherent pattern (UniProt O75556: secreted; standard IHC practice). Predominantly nuclear staining, staining confined to tissue edges or necrosis, and color reproduced without primary antibody point toward artifact or endogenous enzyme activity (HPA subcellular: vesicles; standard chromogenic IHC practice). The A07927 caption documents paraffin-section staining in breast carcinoma but does not establish fixation conditions or diagnostic specificity for a new specimen (selected A07927 tissue-IHC caption).
Boster reagents
Best SCGB2A1 / Mammaglobin-B IHC Antibodies
Two anti-SCGB2A1 antibodies have paraffin-section human breast carcinoma IHC images; both list IF, but neither provides an IF image (catalog image captions and applications).
Real IHC data
Anti-Mammaglobin B (K23) SCGB2A1 Antibody
Cat # A07927
Real IHC data
Anti-Mammaglobin B SCGB2A1 Antibody
Cat # A07927-1
A07927 has an IHC image from paraffin-embedded human breast carcinoma and lists IHC/IF reactivity with human, mouse and rat (A07927 catalog). A07927-1 has a paraffin-embedded human breast carcinoma IHC image with a peptide-blocked comparison and lists IHC/IF reactivity with human and rat (A07927-1 catalog).
Which to pick: For human tissue IHC, consider A07927-1 because its own paraffin-section image includes a peptide-blocked comparison (A07927-1 image caption). For IF, both list the application, but neither provides an IF image; ICC validation is unreported (catalog applications and image captions). For mouse samples, choose A07927 because only it lists mouse reactivity; both antibodies are rabbit-hosted, their clonality is unreported, and neither IHC caption reports the fixative (catalog reactivity, host and clone fields; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75556 (SG2A1_HUMAN, Mammaglobin-B).
- Human Protein Atlas. SCGB2A1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SCGB2A1 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. SCGB2A1 antibody validation summary (1 antibodies).
- Decreased secretoglobin family 2A member 1expression is associated with poor outcomes in endometrial cancer. Oncology letters 2020 — PMC7406884.
- The LINC00365/SCGB2A1 (Mammaglobin B) Axis Down-Regulates NF-κB Signaling and Is Associated with the Progression of Gastric Cancer. Cancer management and research 2020 — PMC6995300.
- Mammaglobin B (SCGB2A1) is a novel tumour antigen highly differentially expressed in all major histological types of ovarian cancer: implications for ovarian cancer immunotherapy. British journal of cancer 2013 — PMC3721400.
- The cytokine-driven regulation of secretoglobins in normal human upper airway and their expression, particularly that of uteroglobin-related protein 1, in chronic rhinosinusitis. Respiratory research 2011 — PMC3063214.
- PubMed PMID:9806831 — UniProt-cited evidence.
- PubMed PMID:10066439 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SCGB2A1 (Mammaglobin-B). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07927 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A07927); 2. Fixation: Image fixative and duration unreported (datasheet A07927); 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-SCGB2A1, 1:50-1:200 (datasheet A07927); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: RNA and protein locations may differ for this secreted protein (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 19–95 (UniProt); Regulation: May be regulated by steroid hormones (UniProt). Evidence: UniProt O75556, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet A07927, PMC6995300. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SCGB2A1/immunohistochemistry-ihc-guide.