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CARD11 / Caspase recruitment domain-containing protein 11 · IHC design guide
Design Immunohistochemistry for CARD11
Plan CARD11 IHC-P with the catalog antibody’s starting concentration of 10 μg/mL (datasheet: A00740). Assess cytoplasmic staining in lymphoid cell subsets, using lymph node, tonsil, spleen or appendix as positive tissue references (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CARD11 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 | Cytoplasm in subsets of lymphoid cells (HPA tissue IHC) | |
| Staining pattern | Subset lymphoid cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+3 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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining was observed and disregarded (HPA tissue IHC) | |
| Regulation | No expression change specified (UniProt) | |
| Isoform / epitope | 0 isoforms; one full-length chain; epitope site unreported (UniProt) |
Section 1
Recommended CARD11 IHC & IF Protocols
The catalog antibody IHC-P protocol is accompanied by published CARD11 methods for kidney cancer tissue microarrays (PMC8568653) and avian organ sections (PMC9969321).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A00740); 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-CARD11, 10 μg/mL (datasheet A00740) |
| 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 | CARD11-positive staining in lymphoid tissue of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in subsets of lymphoid cells in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); the kidney microarray method also uses citrate pH 6.0 (PMC8568653).
Section 2
What Is the Expected CARD11 Staining Pattern?
CARD11 should stain the cytoplasm of subsets of lymphoid cells in paraffin sections (HPA tissue IHC: Approved; cytoplasmic expression in lymphoid subsets). UniProt also places CARD11 in membrane rafts and reports no transmembrane segment, so a continuous cell-surface outline is not required (UniProt Q9BXL7: subcellular location and topology). HPA notes presumed off-target binding, which should temper interpretation of unexpected staining (HPA tissue IHC: reliability description).
| Cytoplasmic staining in some germinal-center cells of lymph node or tonsil, with unstained neighboring cells. | This fits the reported cell pattern. HPA grades germinal-center staining Medium in both tissues and describes expression in subsets of lymphoid cells; a mixed positive and negative field is therefore plausible (HPA tissue IHC: lymph node, tonsil, profile). |
| A strong, uniform nuclear signal dominates cells expected to contain CARD11. | Treat this as an unexpected compartment pattern and check controls and morphology before scoring it as CARD11. The supplied localization is cytoplasm and membrane raft; HPA describes cytoplasmic tissue staining, without a nuclear pattern (UniProt Q9BXL7: subcellular location; HPA tissue IHC: profile). |
| Broad staining appears in a cell population reported as unstained, such as adipocytes in adipose tissue. | Consider nonspecific binding or detection-system activity, then inspect the negative control. HPA reports adipocytes as Not detected and explicitly notes presumed off-target binding in its antibody assessment; a positive reaction here needs independent support (HPA tissue IHC: adipose tissue, reliability description). |
| Diffuse color covers tissue and empty spaces without resolving individual cytoplasms. | This is background until controls show a cell-associated pattern. Review blocking, washes, chromogen development and the no-primary control as general IHC practice. HPA's interpretable CARD11 pattern is cytoplasmic staining in lymphoid subsets, not a field-wide haze (HPA tissue IHC: profile). |
| No cytoplasmic signal is seen in lymph node germinal-center cells or spleen white-pulp cells. | First confirm that the relevant cells are present and that the run worked with a suitable positive control. HPA reports Medium staining in these specific populations, but its Approved rating is not a guarantee that every section or run will show signal (HPA tissue IHC: lymph node, spleen, reliability). |
💡Expected CARD11 appearanceCall a result consistent with CARD11 when Medium cytoplasmic staining is resolved in lymphoid subsets, especially germinal-center or white-pulp cells; dominant nuclear, uniform cell-surface, or broad nonlymphoid staining warrants review (HPA tissue IHC: profile, positive tissues; UniProt Q9BXL7: subcellular location and topology).
| Cell population and tissue selection | Appendix lymphoid tissue, lymph node and tonsil germinal centers, and spleen white pulp are reported at Medium; score the named cells rather than the entire section (HPA tissue IHC: positive tissues, profile). |
| Subcellular context | Cytoplasm is the primary expected compartment. Membrane-raft association can contribute localized signal, but the record gives no IHC requirement for a membrane rim (UniProt Q9BXL7: subcellular location, topology; HPA tissue IHC: profile). |
| Strength of antibody evidence | The listed antibody HPA052984 has IHC status Approved, and HPA reports presumed off-target binding. Unexpected staining needs control-based interpretation; the supplied record does not label this antibody IHC Enhanced (HPA antibodies: HPA052984; HPA tissue IHC: reliability description). |
| Activation-related organization | UniProt describes CARD11 polymerization after T-cell or B-cell receptor activation and a membrane-raft complex. These facts explain possible spatial heterogeneity, but the supplied IHC observations do not establish a reproducible activation-specific stain pattern (UniProt Q9BXL7: function, subunit; HPA tissue IHC: profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive lymphoid tissue is blank. | The relevant cell population may be absent from the section, or the IHC run may have failed; the record gives no CARD11-specific retrieval or dilution setting (HPA tissue IHC: positive tissues). | Confirm germinal-center or white-pulp cells on morphology, check a concurrently stained positive control, then review retrieval and detection against the validated IHC workflow as general IHC practice. |
| A faint reaction is hard to distinguish from background. | HPA reports staining in subsets at Medium, so the expected signal is cell-restricted; diffuse color cannot be assigned to CARD11 from intensity alone (HPA tissue IHC: profile, positive tissues). | Compare the same cell population with a no-primary control and assess cytoplasmic localization at higher magnification; review blocking and wash steps as general IHC practice. |
| Adipocytes or another reported negative population stain strongly. | HPA reports adipocytes as Not detected and acknowledges presumed off-target binding; endogenous detection activity or nonspecific antibody binding are general IHC possibilities (HPA tissue IHC: adipose tissue, reliability description). | Run no-primary and detection-only controls, inspect where the color develops, and seek an independent antibody or orthogonal result before assigning the signal to CARD11 (general IHC practice). |
| Signal appears predominantly nuclear or as a continuous surface rim. | Neither pattern is the reported tissue appearance. UniProt places CARD11 in cytoplasm and membrane rafts and lists no transmembrane segment (HPA tissue IHC: profile; UniProt Q9BXL7: subcellular location, topology). | Check compartment assignment against the counterstain and cell boundaries, then compare a positive lymphoid control; do not score the unexpected pattern as confirmed CARD11 without independent support (general IHC practice). |
| All cells in a lymphoid region stain equally. | HPA describes cytoplasmic expression in subsets of lymphoid cells, so uniform staining may reflect background or an overdeveloped reaction; the record does not define a universal positive-cell fraction (HPA tissue IHC: profile). | Recheck exposure to chromogen, washes and no-primary control as general IHC practice; score identifiable cells and compartments rather than the overall darkness of the region. |
| Can an ICC-IF image or IF protocol be inferred from this IHC pattern? | HPA lists no ICC-IF image-bearing cell lines or subcellular location call for CARD11, and the listed antibody has no ICC validation status (HPA subcellular: image list, location; HPA antibodies: HPA052984). | Use this section only to set a cautious localization expectation. Consult the separate IF/ICC guide for assay design; this IHC evidence does not establish an IF staining result or protocol (HPA tissue IHC: profile; HPA subcellular: image list). |
Experiment evidence
IHC & IF images showing CARD11 staining
A00740 · IHCImmunohistochemistry of CARMA1 in mouse thymus with CARMA1 antibody at 10 μg/mL.
A00740 · IFImmunofluorescence of CARMA1 in Mouse Spleen cells with CARMA1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198286-CARD11/tissue/cerebral+cortex#imid_15699715
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198286-CARD11/tissue/colon#imid_15699625
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198286-CARD11/tissue/liver#imid_15699626
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000198286-CARD11/tissue/kidney#imid_15699635
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CARD11 IHC & IF
🧪Run tonsil first: germinal center cells should stain (HPA: Medium in tonsil germinal center cells). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected); compare cells outside the tonsil germinal centers as internal background without assuming they are all CARD11-negative (HPA: staining reported for germinal center cells).
Positive control tissue: Appendix (Lymphoid tissue, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for CARD11; derive a cell-line control from the positive tissue's cell type (Lymphoid tissue) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and primary host-species- and immunoglobulin-class-matched isotype controls, plus CARD11-knockout material or a peptide-block control where available (standard IHC practice). For chromogenic staining of tonsil, block endogenous peroxidase and assess background in the controls (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or retrieval dependence is reported in the supplied evidence; optimize antigen retrieval empirically for paraffin sections. The A00740 mouse-thymus IHC caption reports 10 µg/mL but leaves the fixative unreported (A00740 caption). There is no evidence here that frozen sections or IF/ICC are easier; tonsil leukocytes can contribute endogenous peroxidase background in chromogenic IHC (standard IHC practice).
HPA tissue IHC evidence for CARD11
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Presumed off target binding observed and disregarded. 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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in white pulp | Medium | Protein (IHC) | HPA → |
| Tonsil | Germinal center 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CARD11 IHC Tips
Troubleshoot CARD11 staining in paraffin sections by checking retrieval, cell identity, cytoplasmic localisation, and controls before comparing staining intensity.
What retrieval conditions should I try first for weak CARD11 staining?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 minutes (page retrieval setting). Run a known positive lymphoid section alongside the test section; germinal center cells in lymph node and tonsil show medium staining (HPA: tissue IHC). If staining remains weak, compare a longer exposure to the same buffer with an EDTA buffer at pH 9, changing only one condition at a time (standard IHC practice). Check whether improved signal remains cytoplasmic in lymphoid cells (HPA: tissue IHC profile), since stronger staining outside that pattern may indicate retrieval-related background.
Could fixation explain weak or uneven CARD11 staining?
Target-specific CARD11 sensitivity to fixation is unknown from the supplied evidence, so assess fixation as a general source of IHC variation (selected A00740 caption: fixative not stated). Record the fixative, time before fixation, and fixation duration for each specimen; compare sections processed under the same conditions (standard IHC practice). If a specimen was fixed in 10% neutral buffered formalin, compare it with a similarly processed positive lymphoid control before changing retrieval (standard IHC practice). Uneven signal near tissue edges or poorly preserved areas warrants a processing review before a biological conclusion, even when the expected lymphoid cells are present (standard IHC practice; HPA: tissue IHC profile).
Where should convincing CARD11 staining appear in a paraffin section?
Expect predominantly cytoplasmic staining in subsets of lymphoid cells (HPA: tissue IHC profile), with possible membrane raft association (UniProt Q9BXL7: subcellular location). CARD11 has no transmembrane segment (UniProt Q9BXL7: topology), so a continuous cell-surface rim alone should prompt a specificity check. Compare stained cells with adjacent morphology and, where available, a serial section carrying a lymphoid marker (standard IHC practice); germinal center cells in lymph node and tonsil provide relevant reference patterns (HPA: tissue IHC). Do not score isolated nuclear staining as CARD11 localisation without independent support, particularly if the same pattern appears in a no-primary control (standard IHC practice; UniProt Q9BXL7: subcellular location).
How should I assess an unexpected staining pattern when the antibody epitope is unclear?
First confirm the antibody’s stated immunogen or epitope and species reactivity in its documentation; the selected tissue caption identifies A00740 but does not define an epitope (selected A00740 caption). The supplied record lists 0 isoforms and a single 1–1154 chain, so an isoform-specific explanation is unsupported here (UniProt Q9BXL7: isoforms and processing). CARD11 contains CARD, PDZ, and guanylate kinase-like domains and several modified residues (UniProt Q9BXL7: domains and modified residues); their effects on this antibody’s staining are unknown. Compare retrieval conditions and a second antibody with a documented, distinct epitope if available, retaining the same positive and negative section controls (standard IHC practice).
How can IF help check the cell identity and location seen by chromogenic IHC?
Use IF as a separate validation experiment and pair CARD11 with a marker identifying the lymphoid population under study; IHC shows cytoplasmic staining in subsets of lymphoid cells (HPA: tissue IHC profile). Choose spectrally separated fluorophores and inspect unstained tissue in each channel before assigning faint signal, because tissue autofluorescence can mimic staining (standard IF practice). Apply permeabilisation for an intracellular epitope: CARD11 is cytoplasmic and has no transmembrane segment (UniProt Q9BXL7: subcellular location and topology). The supplied HPA subcellular record has 0 cell lines with ICC/IF images (HPA: subcellular record), so establish IF antibody performance with appropriate single-stain and no-primary controls (standard IF practice).
What should I check when CARD11 staining is widespread or diffuse?
Check a no-primary control, endogenous peroxidase blocking, secondary reagent concentration, and wash steps before interpreting widespread DAB signal (standard chromogenic IHC practice). HPA reports cytoplasmic expression in subsets of lymphoid cells and notes presumed off-target binding in its reliability assessment (HPA: tissue IHC profile and reliability). Diffuse signal in unrelated cell populations therefore needs confirmation with tissue morphology and an independently validated control (standard IHC practice; HPA: tissue IHC profile). If background rises after retrieval, restore the initial citrate condition of pH 6.0, 95–98 °C, and 20 minutes, then adjust antibody concentration or blocking in separate comparisons (page retrieval setting; standard IHC practice).
How should I quantify CARD11 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: CARD11 staining is cytoplasmic in subsets of lymphoid cells (HPA: tissue IHC profile). Within matched regions, report the percentage of positive target cells and staining intensity, or calculate an H-score from intensity categories weighted by their cell percentages (standard IHC practice). For spatial comparisons, report positive target cells per mm² of viable tissue and normalise counts to the target-cell population or sampled viable area (standard IHC practice). Use identical imaging, threshold, retrieval, and DAB development settings across sections, and exclude folds, necrosis, and tissue edges by a prespecified rule (standard IHC practice).
How can I distinguish true CARD11 signal from staining artefact?
A credible result follows cell identity and compartment: cytoplasmic staining in subsets of lymphoid cells is the reported pattern (HPA: tissue IHC profile). Medium staining in germinal center cells of lymph node and tonsil, or cells in splenic white pulp, offers a reference, while HPA also cautions that presumed off-target binding was observed (HPA: tissue IHC and reliability). Question isolated nuclear staining, a continuous surface rim, or strong staining in an unexpected cell population; CARD11 is cytoplasmic, associates with membrane rafts, and lacks a transmembrane segment (UniProt Q9BXL7: subcellular location and topology). Exclude edge staining, necrosis, and endogenous peroxidase signal with morphology and control sections before interpreting a difference biologically (standard IHC practice).
Boster reagents
Best CARD11 / Caspase recruitment domain-containing protein 11 IHC Antibodies
A00740 has mouse thymus IHC and mouse spleen cell IF images (catalog image captions); human and mouse reactivity is listed (catalog: reactivity).
Real IHC data
Anti-CARMA1 CARD11 Antibody
Cat # A00740
The rendered card is A00740, listed for IHC-P and IF (catalog: applications). Its images show mouse thymus IHC at 10 μg/mL and mouse spleen cell IF at 20 μg/mL (catalog image captions).
Which to pick: Choose A00740 for paraffin-section IHC because IHC-P is listed and its own IHC image shows mouse thymus staining (catalog: applications; A00740 IHC image caption); the fixative is unreported (A00740 IHC image caption). For IF/ICC, A00740 has an IF image of mouse spleen cells, while M00740 is listed for ICC/IF without a supplied image (catalog: A00740 IF image caption; M00740 applications/image alts). Both list human and mouse reactivity, and M00740 is a rabbit monoclonal; only A00740 lists IHC-P (catalog: reactivity/host/clone/applications).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BXL7 (CAR11_HUMAN, Caspase recruitment domain-containing protein 11).
- Human Protein Atlas. CARD11 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CARD11 subcellular location (ICC-IF): Highest expression in RT-4: 72.9 nTPM.
- Human Protein Atlas. CARD11 antibody validation summary (1 antibodies).
- [Expression and prognostic value of CARD11 in diffuse large B cell lymphoma]. Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi 2016 — PMC7342294.
- Gene Body Methylation of the Lymphocyte-Specific Gene CARD11 Results in Its Overexpression and Regulates Cancer mTOR Signaling. Molecular cancer research : MCR 2021 — PMC8568653.
- Enforced CARD11/MALT1 signaling in dendritic cells triggers hemophagocytic lymphohistiocytosis. Proceedings of the National Academy of Sciences of the United States of America 2024 — PMC11665875.
- The brain-specific upregulation of CARD11 in response to avian brain-neurotropic virus infection serves as a potential biomarker. Poultry science 2023 — PMC9969321.
- PubMed PMID:11278692 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
- PubMed PMID:12690205 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CARD11 (Caspase recruitment domain-containing protein 11). Expected localisation Cytoplasm in subsets of lymphoid cells (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A00740 at 10 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A00740); 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-CARD11, 10 μg/mL (datasheet A00740); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining was observed and disregarded (HPA tissue IHC); Isoform / epitope: 0 isoforms; one full-length chain; epitope site unreported (UniProt); Regulation: No expression change specified (UniProt). Evidence: UniProt Q9BXL7, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in RT-4: 72.9 nTPM, Datasheet A00740, PMC8568653, PMC9969321. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CARD11/immunohistochemistry-ihc-guide.