This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
Plan paraffin-section MZB1 IHC with spleen or tonsil as positive tissue controls (HPA tissue IHC; datasheet A08281-1). Assess cytoplasmic staining in immune-cell subsets, accounting for possible differences between protein staining and RNA location (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasm in a subset of immune cells (HPA tissue IHC) | |
| Staining pattern | Subset of immune cells shows cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08281-1) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep tissue fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Secretion may separate protein staining from RNA location (HPA tissue IHC) | |
| Regulation | Higher expression in lymphoid tissue (UniProt) | |
| Isoform / epitope | 5 isoforms; signal peptide 1–22 is removed; check epitope (UniProt) |
The catalog antibody has an EDTA pH 8.0 IHC protocol (datasheet A08281-1). Published MZB1 IHC protocols below cover lymphoid and other tissues (PMC5791339; PMC7491119; PMC6008462).
| Sample | Paraffin-embedded human spleen tissue; fixative not specified (datasheet A08281-1) |
| Fixation | Image fixative and duration unreported (datasheet A08281-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A08281-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08281-1) |
| Primary antibody | Rabbit anti-MZB1, 1-2 μg/ml (datasheet A08281-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08281-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08281-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MZB1-positive staining in germinal center cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in a subset of immune cells in most tissues. No signal in the no-primary control. |
MZB1 is an ER-lumen protein with no transmembrane segment; UniProt also reports secretion and granular cytoplasm around the nucleus (UniProt Q8WU39). In paraffin-section IHC, expect cytoplasmic staining in a subset of immune cells across many tissues (HPA tissue IHC: Enhanced). Interpret the stained cells alongside tissue architecture: a positive tissue does not imply that every cell should stain (HPA tissue IHC).
| Discrete cytoplasmic staining in selected cells of a lymphoid tissue. | This fits the reported IHC profile. HPA records medium staining in germinal center cells of appendix, lymph node and tonsil, and in spleen red-pulp cells (HPA tissue IHC). Score the proportion and intensity of the relevant cells; avoid assigning the tissue one uniform result when neighboring cells differ (general IHC practice). |
| Predominantly nuclear staining, with little convincing cytoplasmic signal. | An isolated nuclear pattern lacks support from the reported ER-lumen and perinuclear cytoplasmic locations (UniProt Q8WU39) or the HPA tissue IHC profile (HPA tissue IHC). Treat it as suspect and compare control sections before calling it MZB1. A Golgi result reported by ICC-IF does not establish nuclear staining in paraffin IHC (HPA subcellular ICC-IF). |
| Strong staining throughout epithelial cells or adipocytes instead of a subset of immune cells. | This differs from the reported IHC distribution (HPA tissue IHC). Adipocytes in adipose tissue and respiratory epithelial cells in bronchus are listed as not detected (HPA tissue IHC). Check for nonspecific antibody binding or endogenous detection activity using appropriate controls (general IHC practice); the image alone cannot distinguish those causes. |
| Uniform, low contrast color over cells and surrounding tissue. | A widespread haze obscures the subset-of-immune-cells pattern reported by HPA (HPA tissue IHC). It cannot establish a positive cell population. Compare the no-primary control, review blocking and washes, and score only signal with a clear cellular boundary above background (general IHC practice). Secreted MZB1 alone does not justify calling uniform chromogen deposition specific (UniProt Q8WU39). |
| No detectable staining in a section expected to contain positive immune cells. | Absence is inconclusive until tissue sampling and assay controls are checked. HPA reports medium staining in specified cells of appendix, bone marrow, lymph node, spleen, thymus and tonsil, not in every cell of those tissues (HPA tissue IHC). Confirm that the relevant compartment is present, then review antibody and detection performance (general IHC practice). |
| Cellular compartment | MZB1 is reported in the ER lumen, secreted space and cytoplasm, with granular perinuclear cytoplasm described (UniProt Q8WU39). Its signal peptide spans residues 1–22, and the annotated mature chain spans 23–189; there is no transmembrane segment (UniProt Q8WU39). These annotations support cytoplasmic interpretation but do not specify an IHC retrieval condition. |
| Which cells are sampled | HPA describes cytoplasmic expression in a subset of immune cells in most tissues, with medium staining in its listed positive cell groups (HPA tissue IHC). Evaluate the named cells within the section rather than treating an unstained neighboring population as assay failure (general IHC practice). |
| Secreted protein and RNA comparison | MZB1 is annotated as secreted (UniProt Q8WU39). HPA warns that tissue locations of RNA and protein may differ for secreted proteins, despite its Enhanced tissue IHC reliability assessment (HPA tissue IHC). Use observed cell-level staining and controls when interpreting a section; RNA distribution alone cannot assign a stained cell (general IHC practice). |
| Antibody validation and isoforms | Both listed antibodies, HPA043745 and HPA052694, have Enhanced IHC status; HPA052694 also has Approved ICC status (HPA antibodies). UniProt lists five MZB1 isoforms (UniProt Q8WU39). These records do not say which isoforms each antibody detects, so do not infer isoform-specific staining from the pattern. |
| IF/ICC Q&A: must its compartment match IHC? | HPA reports Golgi apparatus localization in ICC-IF, with Approved status, while its tissue IHC summary is cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). UniProt reports ER-lumen and perinuclear cytoplasmic localization (UniProt Q8WU39). Interpret each assay against its own evidence; the ICC-IF record supplies no paraffin-section protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known-positive lymphoid section is blank. | The relevant stained cells may be absent from the sampled area, or the IHC workflow may have failed (HPA tissue IHC; general IHC practice). | Locate the expected cell population first; HPA lists medium staining in germinal center cells of lymph node and tonsil, among other sites (HPA tissue IHC). Then check a positive control and the antibody and detection steps (general IHC practice). |
| Color appears mainly in nuclei. | That compartment is unsupported by the supplied MZB1 localization records (UniProt Q8WU39; HPA tissue IHC). | Compare with a no-primary control, inspect cytoplasmic staining in an expected positive cell group, and review counterstain and detection conditions (general IHC practice). Do not score nuclear color alone as MZB1. |
| Adipocytes or bronchial epithelium stain strongly. | HPA lists adipocytes in adipose tissue and respiratory epithelial cells in bronchus as not detected (HPA tissue IHC). Nonspecific binding or endogenous detection activity is possible (general IHC practice). | Run an appropriate no-primary control and inspect the detection chemistry; confirm whether the color follows cells rather than tissue-wide background (general IHC practice). |
| A diffuse chromogenic haze covers the section. | Background from blocking, washes or detection can hide the restricted cellular pattern (general IHC practice; HPA tissue IHC). | Compare a no-primary control, review blocking and wash steps, and assess only clearly bounded cellular signal (general IHC practice). Do not use diffuse extracellular color alone to identify MZB1-positive cells. |
| Only scattered cells stain within a positive tissue. | Restricted staining may be expected: HPA describes a subset of immune cells, and its listed positive cell groups have medium staining (HPA tissue IHC). | Identify the stained cell population and record its fraction and intensity within the relevant area (general IHC practice). Compare with the HPA cell-level pattern before adjusting assay conditions (HPA tissue IHC). |
| IHC looks cytoplasmic while ICC-IF appears Golgi-localized. | The supplied sources report cytoplasmic tissue IHC and Approved Golgi localization in ICC-IF (HPA tissue IHC; HPA subcellular ICC-IF). | Judge the paraffin IHC section against its tissue cell pattern and controls (HPA tissue IHC; general IHC practice). Treat the ICC-IF result as assay-specific evidence, not as a paraffin IHC protocol or a reason by itself to reject cytoplasmic staining. |
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. 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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Medium | Protein (IHC) | HPA → |
| Thymus | Cortical cells | Medium | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Use the paraffin-section IHC evidence for A08281-1 as the starting point, then assess staining by cell type, compartment and controls.
The IHC-validated anti-MZB1 antibody has paraffin-section images from human spleen and tonsil (A08281-1 IHC captions), plus IF data from Jurkat cells (A08281-1 IF caption).
A08281-1 will render with its human spleen IHC figure (A08281-1 card caption). Its additional captions show human tonsil IHC and Jurkat-cell IF (A08281-1 image captions).
Which to pick: Choose A08281-1 for human paraffin-section IHC: its spleen caption reports EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (A08281-1 IHC caption); the fixative is unreported (A08281-1 IHC captions). The same SKU is listed for IF and ICC, with a Jurkat-cell IF image at 5 μg/ml (A08281-1 applications and IF caption). No cross-species choice is supported because the catalog lists human reactivity only (A08281-1 reactivity).