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- Table of Contents
Plan CASP7 IHC in paraffin sections around the cytoplasmic and additional nuclear staining observed in tissue (HPA tissue IHC). Compare glandular cells in the gastrointestinal tract, where staining is most abundant (HPA tissue IHC), and interpret signal in light of CASP7 cleavage (UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasmic, with additional nuclear staining (HPA tissue IHC) | |
| Staining pattern | GI glandular cells: cytoplasmic with some nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01044-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining alone does not establish CASP7 activation (UniProt) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 4 isoforms; cleavage yields p20/p11, so epitope choice matters (UniProt) |
Compare the catalog antibody’s IHC-P protocol (datasheet A01044-2) with published CASP7 IHC methods for human lung cancer cases (PMC4673127) and porcine endometrium (PMC7907442).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01044-2) |
| Fixation | Image fixative and duration unreported (datasheet A01044-2); 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 A01044-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01044-2) |
| Primary antibody | Rabbit anti-CASP7, 2-5μg/ml (datasheet A01044-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01044-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01044-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CASP7-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic with additional nuclear expression, most abundant in gastrointestinal tract. No signal in the no-primary control. |
CASP7 is a soluble protein found in the cytosol and nucleus, with no transmembrane segment (UniProt P55210). On paraffin sections, expect predominantly cytoplasmic staining with some nuclear staining, especially in gastrointestinal glandular cells (HPA tissue IHC). HPA rates the tissue IHC profile Enhanced, while describing only medium consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining, sometimes accompanied by nuclear staining, is strongest in appendix, colon, duodenum, rectum or stomach glands. | This fits the reported tissue pattern: these glandular cells are High, and the overall profile is generally cytoplasmic with additional nuclear expression (HPA tissue IHC). Compare like cell types within a section; intensity in neighboring cells need not be uniform (standard IHC practice). |
| Staining appears exclusively as a sharp cell-border rim, with little cytoplasmic or nuclear signal. | Treat a membrane-only IHC pattern as discordant with the reported tissue profile and check for artefact (HPA tissue IHC; standard IHC practice). Do not dismiss every membrane-associated observation: HPA reports an additional plasma-membrane location by ICC-IF, which is a different assay (HPA subcellular ICC-IF). |
| Strong staining dominates adipocytes, skeletal myocytes or cardiomyocytes while gastrointestinal glands are weak. | Those cell types are listed as Not detected, whereas several gastrointestinal glandular populations are High (HPA tissue IHC). Suspect nonspecific staining, cross-reactivity or detection background; assess the tissue architecture and controls before assigning biological meaning (standard IHC practice). |
| A diffuse tint covers cells, stroma and blank areas without a discernible intracellular pattern. | This does not resemble the reported cytoplasmic-plus-nuclear tissue profile (HPA tissue IHC). In chromogenic IHC, widespread tint can reflect nonspecific reagent binding or endogenous detection activity; inspect a no-primary control before scoring it as CASP7 (standard IHC practice). |
| No staining is seen in well-preserved gastrointestinal glandular cells. | Investigate the assay before calling the sample CASP7-negative: appendix, colon, duodenum, rectum and stomach glandular cells are reported High (HPA tissue IHC). Confirm the expected cells are present, then review positive-control performance and the antibody’s IHC validation (HPA tissue IHC; standard IHC practice). |
| Tissue and cell identity | HPA reports High staining in several gastrointestinal glandular populations, Medium in bronchial respiratory epithelium and cervical glands, and Low in liver hepatocytes and kidney tubules (HPA tissue IHC). Score the named cell population rather than the whole organ; weak staining in a Low population is compatible with the atlas profile (HPA tissue IHC). |
| Antibody validation and assay | CAB025563 has an Enhanced IHC validation status; HPA069167 is listed as Supported for ICC without an IHC status (HPA antibodies). Apply the IHC evidence to paraffin-section interpretation, and keep ICC-IF localization observations separate when comparing compartments (HPA antibodies; HPA subcellular ICC-IF). |
| Processing and epitope identity | CASP7 is processed into p20 and p11 chains, and activated CASP7 can be secreted through GSDMD or perforin pores in specified settings (UniProt P55210). Whether a stain detects precursor, cleaved CASP7 or both depends on an antibody’s epitope; no epitope information is supplied here (UniProt P55210 processing; antibody information supplied). Do not interpret intensity alone as activation (standard IHC practice). |
| Isoforms and tissue-source disagreement | UniProt lists four CASP7 isoforms and reports no brain expression, while HPA reports High staining in cerebellar Purkinje cells (UniProt P55210; HPA tissue IHC). This disagreement makes Purkinje staining a finding to verify with the chosen antibody, rather than an uncontested positive reference (HPA tissue IHC; standard IHC practice). |
| Detection chemistry | Endogenous enzyme activity or nonspecific chromogen deposition can complicate chromogenic IHC interpretation (standard IHC practice). Evaluate detection controls and any activity block according to the detection system used; neither HPA nor UniProt supplies a CASP7-specific blocking requirement (standard IHC practice; HPA tissue IHC; UniProt P55210). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected gastrointestinal glands are unstained. | A missing signal may reflect an assay or tissue-control problem; these glandular populations are reported High (HPA tissue IHC; standard IHC practice). | Verify glandular cells on the section and review the IHC-validated antibody, retrieval and detection controls before scoring the sample negative (HPA antibodies; standard IHC practice). No CASP7-specific retrieval setting is supplied (HPA tissue IHC; UniProt P55210). |
| Adipocytes or muscle stain more strongly than the expected glands. | HPA lists adipocytes, skeletal myocytes and cardiomyocytes as Not detected, so this distribution warrants a specificity check (HPA tissue IHC). | Compare with the no-primary control and an expected-positive glandular area; assess whether the signal follows cell boundaries, tissue debris or the intended intracellular pattern (standard IHC practice; HPA tissue IHC). |
| Brown signal extends across blank areas or obscures cell compartments. | Diffuse chromogen can arise from detection background or nonspecific binding (standard IHC practice). | Review no-primary and detection-system controls, then adjust general blocking or detection conditions only as indicated by those controls (standard IHC practice). Reassess whether cytoplasmic and nuclear staining remains distinguishable (HPA tissue IHC). |
| The section shows nuclear staining alongside cytoplasmic staining. | Additional nuclear staining is part of the HPA tissue IHC profile, and UniProt also lists a nuclear location (HPA tissue IHC; UniProt P55210). | Score nuclear and cytoplasmic compartments separately in the relevant cells (standard IHC practice). Investigate only if the pattern lacks the expected cell specificity or resembles diffuse background (HPA tissue IHC; standard IHC practice). |
| Cerebellar Purkinje cells stain, but another brain population does not. | HPA reports High Purkinje-cell staining, whereas UniProt states no brain expression; HPA also lists several glial populations as Not detected (HPA tissue IHC; UniProt P55210). | Record the cell type and this source disagreement. Check antibody specificity and controls before using Purkinje staining as evidence of CASP7 expression (HPA tissue IHC; UniProt P55210; standard IHC practice). |
| IF/ICC Q&A: Should a nucleolar or plasma-membrane IF signal be scored as the expected paraffin IHC pattern? | HPA’s ICC-IF summary places CASP7 mainly in the nucleoplasm and nucleoli fibrillar center, with an additional plasma-membrane location; the tissue IHC summary emphasizes cytoplasmic and additional nuclear staining (HPA subcellular ICC-IF; HPA tissue IHC). | Interpret IF/ICC using its own assay evidence and guide. For this paraffin IHC section, use the tissue IHC compartment and cell-type profile to judge the slide (HPA subcellular ICC-IF; HPA tissue IHC). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Rectum | Glandular cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Troubleshoot CASP7 staining in paraffin sections by separating technical signal from cell type, compartment and activation state (UniProt P55210; HPA tissue IHC).
Three anti-CASP7 antibodies have human cancer tissue IHC images (catalog IHC captions); A01044-2 also lists human IF/ICC applications (catalog applications and reactivity), although no IF figure is supplied (catalog IF image alts).
A01044-2 has a paraffin section IHC image from human breast cancer tissue and lists human and rat reactivity (catalog IHC caption; catalog reactivity). PA1442 has a human intestinal cancer IHC(P) image and lists human, mouse and rat reactivity; PB9369 has a human mammary cancer IHC(P) image and lists human and rat reactivity (catalog IHC captions; catalog reactivity).
Which to pick: For tissue IHC, choose A01044-2 when its documented human breast cancer paraffin section procedure is useful: the caption specifies EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and DAB detection, but does not report the fixative (A01044-2 IHC caption). For IF/ICC, choose A01044-2 because it alone lists those applications at 5 μg/ml for human samples; no IF image is supplied (catalog applications, dilution and IF image alts). For planned work across human, mouse and rat, PA1442 has the broadest listed reactivity, although its IHC(P) image shows only human intestinal cancer tissue; its fixative is unreported (catalog reactivity; PA1442 IHC caption).