CASP7 / Caspase-7 · IHC design guide

Design Immunohistochemistry for CASP7

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).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for CASP7 (IHC for CASP7): expected localisation Cytoplasmic, with additional nuclear staining (HPA tissue IHC), antibody A01044-2, validated IHC image, and IHC protocol steps
Printable CASP7 IHC protocol sheet — expected localisation Cytoplasmic, with additional nuclear staining (HPA tissue IHC), antibody A01044-2, controls and protocol steps. Open the full CASP7 IHC guide →

CASP7 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, 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
Important caveats
Reasons your staining may differ from the expected pattern.
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)
Section 1

Recommended CASP7 IHC & IF Protocols

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).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human breast cancer tissue; fixative not specified (datasheet A01044-2)
FixationImage fixative and duration unreported (datasheet A01044-2); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A01044-2); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A01044-2)
Primary antibodyRabbit anti-CASP7, 2-5μg/ml (datasheet A01044-2)
Primary incubationOvernight at 4 °C (datasheet A01044-2)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet A01044-2)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultCASP7-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.
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A01044-2); the porcine study used citrate at pH 6.0 (PMC7907442).
Section 2

What Is the Expected CASP7 Staining Pattern?

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).

What am I looking at on my slide?
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).
💡Expected CASP7 appearanceCall a positive result when gastrointestinal glandular cells show conspicuous cytoplasmic staining, with possible nuclear staining; a membrane-only rim or uniform stain across blank areas is suspect (HPA tissue IHC; standard IHC practice).
How each factor affects the staining
Tissue and cell identityHPA 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 assayCAB025563 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 identityCASP7 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 disagreementUniProt 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 chemistryEndogenous 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).
Why is my staining missing, weak or wrong?
SituationLikely causeNext 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).

Sample controls for CASP7 IHC & IF

🧪Run colon first: its glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes), and treat unstained cells within the colon section as internal background references without assuming a specific cell type is CASP7-negative (HPA: colon glandular-cell staining only).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CASP7 in HEK293, Rh30, U2OS, with annotated localisation: Nucleoplasm (approved), Nucleoli fibrillar center (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and clonality; the catalog primary is rabbit-derived (caption: rabbit anti-CASP7). Confirm specificity with CASP7 knockout material or a validated peptide block, and quench endogenous peroxidase and assess endogenous biotin in the colon section when using the caption’s SABC–DAB detection (caption: SABC–DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01044-2 paraffin-section caption does not state the fixative (caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (caption: EDTA retrieval). No supplied comparison establishes whether frozen sections or IF are easier; in colon, check glandular luminal material for misleading DAB deposits (standard IHC practice).

HPA tissue IHC evidence for CASP7

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.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
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 →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Section 3

Advanced CASP7 IHC Tips

Troubleshoot CASP7 staining in paraffin sections by separating technical signal from cell type, compartment and activation state (UniProt P55210; HPA tissue IHC).

Which retrieval conditions should I try first for weak CASP7 staining?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01044-2). The selected image used this retrieval before incubation with 2 µg/mL antibody overnight at 4°C; it does not establish an optimal heating time (datasheet A01044-2). If staining remains weak, compare retrieval duration on matched sections while holding antibody concentration and detection constant (standard IHC practice). Include a glandular cell reference section, such as colon, and assess signal in the expected cells rather than judging overall section darkness (HPA tissue IHC: High in colon glandular cells; standard IHC practice).
Could fixation explain weak or uneven CASP7 staining?
CASP7 specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin tissue caption does not state a fixative (datasheet A01044-2). Record the actual fixative and processing history for each section, then compare sections processed together before attributing differences to the target (standard IHC practice). Examine whether weak staining tracks areas with poor preservation, variable section thickness or tissue edges, using morphology and matched controls (standard IHC practice). Keep retrieval at EDTA pH 8.0 during this comparison, because that is the documented condition for this antibody (datasheet A01044-2).
How should I assess cytoplasmic, nuclear or membrane CASP7 staining?
Score cytoplasmic and nuclear staining separately in paraffin sections, since tissue IHC describes general cytoplasmic staining with additional nuclear expression (HPA tissue IHC). CASP7 is also annotated in the cytosol and nucleus, while a separate ICC/IF dataset reports nucleoplasm, nucleolar fibrillar center and additional plasma membrane localisation (UniProt P55210 subcellular location; HPA subcellular). Do not treat membrane staining alone as proof of a membrane spanning protein: CASP7 has no transmembrane segment (UniProt P55210 topology). Check whether an unusual compartmental pattern repeats in appropriate cells and across replicate sections before interpreting it biologically (standard IHC practice).
Can this antibody distinguish CASP7 isoforms or activated CASP7?
The supplied caption establishes tissue staining with A01044-2, but gives no epitope or cleavage specificity, so its chromogenic signal cannot identify an isoform or activated CASP7 (datasheet A01044-2). CASP7 has 4 annotated isoforms, and processing yields the 24–198 and 207–303 chains (UniProt P55210 isoforms and processing). An antibody recognizing a region absent from a processed form could produce a different staining pattern, depending on its mapped epitope (UniProt P55210 processing; standard immunostaining principle). Obtain epitope and validation information before labeling positive cells as cleavage positive, and use a cleavage specific assay when activation is the question (standard IHC practice).
How can IF help resolve ambiguous CASP7 localisation seen by chromogenic IHC?
Use IF as a separate localisation check and multiplex CASP7 with a marker for the cell type whose IHC staining is being assessed; colon glandular cells are a documented high staining reference (HPA tissue IHC; standard IF practice). Choose fluorophores and imaging channels after checking tissue autofluorescence, and include single stain controls to assess channel bleed through (standard IF practice). For an intracellular epitope, assess permeabilisation because CASP7 is annotated in the cytosol and nucleus and lacks a transmembrane segment (UniProt P55210 subcellular location and topology; standard IF practice). Interpret IF compartments against the IHC pattern without assuming that the paraffin tissue caption establishes IF fixation or performance (datasheet A01044-2; HPA subcellular).
What should I check when CASP7 DAB staining is diffuse?
Compare antibody stained sections with a no primary control to identify detection background, and inspect endogenous peroxidase activity before interpreting DAB deposits (standard chromogenic IHC practice). The selected paraffin image used 10% goat serum blocking, biotinylated secondary antibody, streptavidin–biotin detection and DAB, so evaluate background at each relevant detection step (datasheet A01044-2). Diffuse colour across stromal spaces or section edges deserves scrutiny when the intended readout is staining within identifiable cells (standard IHC practice). Use the documented 2 µg/mL primary concentration as the comparison point, then assess any concentration changes with matched controls (datasheet A01044-2; standard IHC practice).
How should I quantify heterogeneous CASP7 staining in paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; report cytoplasmic and nuclear results separately where both appear (HPA tissue IHC; standard IHC practice). An H-score can combine the percentage of cells at each intensity, while percentage positive cells or positive cells per mm² may suit a simpler endpoint (standard IHC practice). Normalise cell counts to the number of evaluable cells, or spatial counts to viable tissue area, and exclude folds, empty spaces and necrotic regions consistently (standard IHC practice). Apply the same positivity threshold and imaging conditions across sections, with a documented glandular cell reference to monitor staining consistency (HPA tissue IHC; standard IHC practice).
When does CASP7 staining support a biological conclusion rather than artefact?
A credible positive follows identifiable cell boundaries, persists away from cut edges and is reproducible in matched sections and controls (standard IHC practice). Compare cell identity with the tissue profile: gastrointestinal glandular cells stain highly, whereas adipocytes and several muscle cell groups are reported as not detected (HPA tissue IHC). Treat colour concentrated at edges, in necrosis or in a no primary control as possible artefact, and check endogenous enzyme activity in DAB sections (standard chromogenic IHC practice). Finally, total CASP7 staining does not by itself establish apoptosis or protease activation, because CASP7 is a processed effector protease and this antibody's cleavage specificity is unspecified (UniProt P55210 function and processing; datasheet A01044-2).
Boster reagents

Best CASP7 / Caspase-7 IHC Antibodies

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).

Real IHC data IHC analysis of Caspase-7/CASP7 using anti-Caspase-7/CASP7 antibody (A01044-2). Caspase-7/CASP7 was detected in paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Caspase-7/CASP7 Antibody (A01044-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Anti-Caspase-7/CASP7 Antibody ®
Cat # A01044-2
Real IHC data Anti-Caspase-7(P11) antibody, PA1442, IHC(P) IHC(P): Human Intestinal Cancer Tissue
Anti-Caspase-7(P11)/CASP7 Antibody ®
Cat # PA1442
Real IHC data Anti-Caspase-7 vendor antibody, PB9369,IHC(P) IHC(P): Human Mammary Cancer Tissue
Anti-Caspase-7/CASP7 Antibody ®
Cat # PB9369

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).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry P55210 (CASP7_HUMAN, Caspase-7).
  2. Human Protein Atlas. CASP7 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. CASP7 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nucleoli fibrillar center. In addition localized to the plasma membrane..
  4. Human Protein Atlas. CASP7 antibody validation summary (2 antibodies).
  5. MicroRNA-224 is implicated in lung cancer pathogenesis through targeting caspase-3 and caspase-7. Oncotarget 2015 — PMC4673127.
  6. Spatiotemporal characterization of extracellular matrix maturation in human artificial stromal-epithelial tissue substitutes. BMC biology 2024 — PMC11575135.
  7. Expression of Caspases in the Pig Endometrium Throughout the Estrous Cycle and at the Maternal-Conceptus Interface During Pregnancy and Regulation by Steroid Hormones and Cytokines. Frontiers in veterinary science 2021 — PMC7907442.
  8. miR-224 Is Significantly Upregulated and Targets Caspase-3 and Caspase-7 During Colorectal Carcinogenesis. Translational oncology 2019 — PMC6240712.
  9. PubMed PMID:8521391 — UniProt-cited evidence.
  10. PubMed PMID:8576161 — UniProt-cited evidence.
  11. PubMed PMID:8567622 — UniProt-cited evidence.