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CAMP / Cathelicidin antimicrobial peptide · IHC design guide
Design Immunohistochemistry for CAMP
Plan CAMP IHC-P with the human-reactive catalog antibody starting at 5 μg/mL (datasheet). Use bone marrow or spleen as positive tissue and assess selective cytoplasmic staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CAMP 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 staining in subsets of immune cells (HPA tissue IHC) | |
| Staining pattern | Selective cytoplasmic immune-cell staining in marrow and spleen (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Bone marrow+2 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 | Secretion can separate tissue protein and RNA signals (HPA tissue IHC) | |
| Regulation | Staining regulation not reported (UniProt) | |
| Isoform / epitope | No isoforms annotated; propeptide is processed (UniProt) |
Section 1
Recommended CAMP IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by one published CAMP IHC protocol for testicular tissue (PMC11559180).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A05475); 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-CAMP, 5 μg/mL (datasheet A05475) |
| 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 | CAMP-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in subsets of immune cells in several tissues, most abundant in bone marrow and spleen. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval); the published protocol uses higher-pressure citrate retrieval (PMC11559180).
Section 2
What Is the Expected CAMP Staining Pattern?
CAMP should appear as selective cytoplasmic staining in subsets of immune cells, strongest in bone marrow hematopoietic cells and spleen red-pulp cells (HPA tissue IHC: Enhanced; High in both). Its secreted, vesicle-associated biology and lack of a transmembrane segment support intracellular granule-associated signal without a required membrane outline (UniProt P49913: subcellular location and topology).
| Strong, selective cytoplasmic staining in bone marrow hematopoietic cells or spleen red-pulp cells (HPA tissue IHC: High in both). | This matches the reported tissue pattern (HPA tissue IHC: Enhanced). Granular cytoplasmic signal is plausible because CAMP is stored in neutrophil granules and phagolysosomes (UniProt P49913: subcellular location); HPA does not establish a required granule pattern. |
| Predominantly nuclear staining or a uniform cell-surface rim, without the expected cytoplasmic signal (HPA tissue IHC: selective cytoplasmic expression). | Treat this as discordant with HPA's observed compartment and UniProt's lack of a transmembrane segment (HPA tissue IHC; UniProt P49913: topology). Review controls and staining conditions before interpreting it as CAMP (standard IHC practice). |
| Strong staining in adipocytes or adrenal glandular cells (HPA tissue IHC: Not detected in those cell types). | This conflicts with the reported cell-type pattern; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; standard IHC practice). Check a primary-omission control and compare with a known-positive tissue before assigning specificity (standard IHC practice). |
| Broad staining across cells and tissue spaces, with little separation between positive and negative areas (HPA tissue IHC: selective cytoplasmic expression). | Diffuse signal is less persuasive than a cell-restricted pattern (HPA tissue IHC). Secreted CAMP can make extracellular signal biologically plausible, so use controls to distinguish it from background (UniProt P49913: secreted; standard IHC practice). |
| No staining in bone marrow hematopoietic cells despite an apparently intact section (HPA tissue IHC: High). | A negative result in this reported positive compartment limits interpretation of other negative samples (HPA tissue IHC). Check antibody and detection controls, then review retrieval and dilution as general IHC variables; CAMP-specific retrieval sensitivity is unreported (standard IHC practice). |
💡Expected CAMP appearanceCall a result positive when selective cytoplasmic signal is strong in bone marrow hematopoietic cells or spleen red-pulp cells (HPA tissue IHC: High); isolated nuclear or widespread cell-type-mismatched staining warrants control review (HPA tissue IHC; standard IHC practice).
| Tissue and cell selection (HPA tissue IHC). | Bone marrow hematopoietic cells and spleen red-pulp cells are reported High; round or early spermatids are Medium (HPA tissue IHC). Score the identified cells, since HPA describes expression as selective within immune-cell subsets (HPA tissue IHC). |
| Secretion and compartment (UniProt P49913; HPA tissue IHC). | CAMP is secreted and stored in neutrophil granules and phagolysosomes (UniProt P49913). HPA observes selective cytoplasmic staining and cautions that RNA and protein tissue locations may differ for a secreted protein (HPA tissue IHC). |
| Processing and antibody epitope (UniProt P49913). | The precursor has a signal peptide at residues 1–30 and propeptide at 31–131; postsecretory processing produces peptides of different lengths (UniProt P49913). Epitope-dependent recognition is possible, but the supplied record does not locate an antibody epitope. |
| IHC evidence and antibody choice (HPA antibodies). | HPA029874, CAB015949 and CAB016522 each have Enhanced IHC validation (HPA antibodies). This supports comparison against HPA's tissue pattern; it does not establish identical staining for every antibody or preparation (HPA antibodies; standard IHC practice). |
| IF/ICC expectation? (HPA subcellular; HPA antibodies). | HPA lists CAMP as Secreted but provides no main ICC-IF location or cell-line images, and no ICC status for the listed antibodies (HPA subcellular; HPA antibodies). An IF/ICC staining pattern therefore cannot be specified from this evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow positive control is blank (HPA tissue IHC: High in hematopoietic cells). | The run may have inadequate primary-antibody signal or failed detection (standard IHC practice); the supplied sources do not identify a CAMP-specific retrieval requirement. | Confirm tissue morphology and positive-control setup, then review antibody dilution, retrieval and detection steps under the assay's general IHC workflow (standard IHC practice). |
| Adipocytes stain strongly (HPA tissue IHC: Not detected in adipocytes). | Cell-type discordance raises concern for nonspecific antibody binding or detection background (HPA tissue IHC; standard IHC practice). | Compare with a primary-omission control and reported positive marrow or spleen cells; reassess specificity if the discordant signal persists (HPA tissue IHC; standard IHC practice). |
| Nuclear staining dominates (HPA tissue IHC: selective cytoplasmic expression). | The compartment conflicts with HPA's cytoplasmic pattern and UniProt's secreted, vesicle-associated annotation (HPA tissue IHC; UniProt P49913). | Inspect negative controls and compare the same run with cytoplasmic signal in a reported positive tissue before scoring CAMP (HPA tissue IHC; standard IHC practice). |
| Chromogenic signal is widespread, including areas outside cells (HPA tissue IHC: selective expression). | Background or endogenous detection activity is possible (standard IHC practice); secretion also makes some extracellular CAMP plausible (UniProt P49913). | Check the primary-omission control and the assay's blocking and detection steps; retain extracellular signal as interpretable only when controls support specificity (standard IHC practice). |
| Only weak signal appears in a reported positive tissue (HPA tissue IHC: High in marrow and spleen). | Low assay sensitivity or sampling of the wrong cell population may explain the mismatch (HPA tissue IHC; standard IHC practice). | Identify hematopoietic or red-pulp cells first, then review retrieval, antibody dilution and detection as general IHC variables; do not infer a CAMP fixation effect (HPA tissue IHC; standard IHC practice). |
| IF/ICC gives a punctate or diffuse pattern that seems unlike IHC (HPA subcellular: no main ICC-IF location). | The supplied HPA record lacks ICC-IF images and antibody ICC status, so it provides no validated IF compartment for comparison (HPA subcellular; HPA antibodies). | Assess IF/ICC with its own controls and guide; use the HPA IHC tissue pattern only as tissue context, not as an IF validation claim (HPA tissue IHC; HPA subcellular; standard IF practice). |
Experiment evidence
IHC & IF images showing CAMP staining
A05475 · IHCImmunohistochemistry of Catheli-cidin in human spleen tissue with Cathelicidin antibody at 5 μg/mL.
A05475 · IHCImmunohistochemistry of Cathelicidin in human spleen tissue with Cathelicidin antibody at 5 μg/ml.
A05475 · IFImmunofluorescence of Cathelicidin in Human Spleen cells with Cathelicidin antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000164047-CAMP/tissue/cerebral+cortex#imid_7466860
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000164047-CAMP/tissue/colon#imid_7434774
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000164047-CAMP/tissue/liver#imid_7434767
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000164047-CAMP/tissue/kidney#imid_7434768
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CAMP IHC & IF
🧪Run bone marrow first and score hematopoietic cells for staining (HPA: High in bone marrow hematopoietic cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the bone marrow slide, assess morphologically identified adipocytes as an internal background check, without treating their staining status as an HPA result for marrow.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for CAMP; derive a cell-line control from the positive tissue's cell type (Hematopoietic cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host and immunoglobulin class; use a CAMP knockout specimen or cognate-peptide competition, if available, to assess target specificity (standard IHC practice). Quench endogenous peroxidase in bone marrow for chromogenic detection, and check unstained tissue for autofluorescence if evaluating IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window, fixation effect, and antigen-retrieval dependency are unreported in the supplied evidence; the fixative for the selected A05475 spleen IHC caption is also unreported (selected-SKU caption). The evidence does not establish whether frozen sections or IF/ICC would be easier for CAMP (HPA: no ICC-IF cell-line images; selected-SKU caption: tissue IHC only). In bone marrow, endogenous peroxidase can complicate chromogenic scoring, so interpret staining against the quenched controls (standard IHC practice).
HPA tissue IHC evidence for CAMP
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.). 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | Protein (IHC) | HPA → |
| Testis | Round or early spermatids | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CAMP IHC Tips
Troubleshoot CAMP staining in paraffin sections by checking retrieval, cell identity, compartment, and controls before comparing signal intensity across specimens.
Which retrieval conditions should I try first for weak CAMP staining?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). Let sections cool consistently, then compare the same tissue across retrieval runs so changes in staining can be attributed to the retrieval step (standard IHC practice). Include a known positive compartment, such as spleen red-pulp cells or bone marrow hematopoietic cells, when judging a weak result (HPA: High). If staining remains weak, adjust heating or try another retrieval condition as a fallback, recording its effect on tissue morphology and background (standard IHC practice). Do not treat the spleen image at 5 µg/mL as evidence for a different retrieval method; its caption reports antibody concentration only (A05475 tissue-IHC caption).
Could fixation explain inconsistent CAMP staining between paraffin blocks?
Target-specific fixation sensitivity is unknown from the supplied evidence, so a weak block cannot be assigned to a particular fixative or fixation duration (A05475 tissue-IHC caption: fixative unreported). Compare blocks with documented, similar processing histories and stain them in the same run before changing the antibody concentration (standard IHC practice). Check whether the positive immune-cell population is present in each section, because CAMP staining is selective across tissues and cells (HPA: tissue IHC profile). If morphology is poor or staining varies across a section, review processing records and section quality before interpreting intensity differences (standard IHC practice). The spleen caption reports 5 µg/mL but gives no fixation condition to reproduce (A05475 tissue-IHC caption).
Where should CAMP signal appear in a positive tissue section?
Expect selective cytoplasmic staining in subsets of immune cells, with strong reported staining in bone marrow hematopoietic cells and spleen red-pulp cells (HPA: tissue IHC profile). CAMP is stored as a propeptide in neutrophil granules and phagolysosomes and can be secreted, so intracellular and extracellular signal require separate interpretation (UniProt P49913: subcellular location). It has no transmembrane segment; a crisp, continuous plasma-membrane outline alone is therefore difficult to reconcile with its annotated topology (UniProt P49913: topology). Review staining at the cellular level against a hematoxylin counterstain, including areas adjacent to strongly positive cells (standard IHC practice). Do not assume protein and local RNA must occupy the same tissue position for this secreted target (HPA: reliability description).
Could epitope placement change which CAMP forms the stain detects?
Map the antibody's stated immunogen or epitope against the 170-aa precursor before interpreting a stain as mature peptide (UniProt P49913: sequence length and processing). The annotated signal peptide spans residues 1–30, and the propeptide spans 31–131; postsecretory processing generates peptides of multiple lengths (UniProt P49913: processing and function). An antibody recognizing a region removed during processing may mark precursor-containing cells without detecting every secreted product (inference from UniProt P49913: processing). No isoforms or glycosylation sites are annotated, so neither provides an evidence-based explanation for divergent staining here (UniProt P49913: isoforms and glycosylation). If epitope information is unavailable, report staining as CAMP immunoreactivity rather than assigning it to one processed form (standard IHC interpretation).
How should I check CAMP localisation in a multiplex IF experiment?
Use a validated neutrophil marker alongside CAMP to test whether intracellular signal occurs in the expected cell population (UniProt P49913: neutrophilic granulocyte expression; standard IF practice). Choose a fluorophore in a channel with low measured tissue autofluorescence, and assess each channel with single-stain and no-primary controls (standard IF practice). CAMP is secreted and stored in granules and phagolysosomes, with no annotated transmembrane segment; if the antibody recognizes an intracellular epitope, test permeabilisation after fixation (UniProt P49913: subcellular location and topology; standard IF practice). If the epitope is unknown, compare permeabilised and nonpermeabilised preparations before assigning a compartment (standard IF practice). The supplied tissue image documents chromogenic IHC at 5 µg/mL, not IF performance (A05475 tissue-IHC caption).
How can I reduce diffuse or misleading chromogenic CAMP background?
Run a no-primary control and inspect whether brown signal persists after the usual endogenous-peroxidase block; residual signal can reflect the detection workflow (standard chromogenic IHC practice). Titrate the catalog antibody around the documented 5 µg/mL spleen-image concentration while holding retrieval, detection, and imaging conditions constant (A05475 tissue-IHC caption; standard IHC practice). Apply an appropriate protein block, wash consistently, and avoid interpreting precipitate or strong section-edge staining as cellular signal (standard IHC practice). Because CAMP is secreted, some extracellular staining may occur, but diffuse color without identifiable positive cells still needs control-based review (UniProt P49913: subcellular location; standard IHC interpretation). Compare with expected spleen red-pulp or bone marrow hematopoietic-cell staining before accepting a broad tissue-wide pattern (HPA: tissue IHC profile).
What should I score when comparing CAMP staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and region of interest before scoring, since CAMP expression is selective within tissues (HPA: tissue IHC profile; standard IHC practice). Record the percentage of positive eligible cells and an intensity-based H-score, or count positive cells per mm² when cell density is the primary outcome (standard IHC practice). Normalize cell-based scores to the number of eligible cells in the same region, and density scores to the analyzed tissue area (standard IHC practice). Keep retrieval, antibody concentration, chromogen development, and image thresholds consistent across the comparison (standard IHC practice). Score intracellular cellular staining separately from extracellular deposits because CAMP can be stored in vesicles and secreted (UniProt P49913: subcellular location).
How do I distinguish true CAMP staining from an artefact?
Prioritize staining in identifiable immune-cell cytoplasm, especially spleen red-pulp cells or bone marrow hematopoietic cells, when judging a positive pattern (HPA: tissue IHC profile). Intracellular granule-associated signal is plausible for neutrophils, whereas a uniform membrane rim is discordant with the annotated absence of a transmembrane segment (UniProt P49913: subcellular location and topology). Inspect section edges and necrotic or damaged areas separately, and compare suspicious color with the no-primary control and peroxidase-block check (standard IHC practice). Extracellular signal can fit a secreted protein but should not, by itself, identify the producing cell (UniProt P49913: subcellular location; standard IHC interpretation). Confirm that the expected cell population and morphology support the conclusion before calling weak or diffuse chromogen true CAMP staining (HPA: tissue IHC profile; standard IHC practice).
Boster reagents
Best CAMP / Cathelicidin antimicrobial peptide IHC Antibodies
A05475 lists human IHC-P and IF applications (catalog: applications and reactivity), with images of human spleen tissue by IHC and human spleen cells by IF (image captions).
Real IHC data
Anti-Cathelicidin CAMP Antibody
Cat # A05475
A05475 lists IHC-P for human samples, with its IHC image showing human spleen tissue at 5 μg/mL (catalog: applications and reactivity; IHC image caption). A05475 also lists IF, with its IF image showing human spleen cells at 20 μg/mL (catalog: applications; IF image caption).
Which to pick: For paraffin-section tissue IHC, choose A05475 based on its IHC-P listing and human spleen tissue image (catalog: IHC-P; IHC image caption); the fixative is unreported (IHC image caption). For IF, choose A05475 based on its human spleen cell image (IF image caption); ICC validation is unreported (catalog: application list). No cross-species choice is supported because A05475 lists Human reactivity only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49913 (CAMP_HUMAN, Cathelicidin antimicrobial peptide).
- Human Protein Atlas. CAMP tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CAMP subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. CAMP antibody validation summary (3 antibodies).
- Nobiletin, as a Novel PDE4B Inhibitor, Alleviates Asthma Symptoms by Activating the cAMP-PKA-CREB Signaling Pathway. International journal of molecular sciences 2024 — PMC11477036.
- Prediction model of male reproductive function damage caused by CHOP chemotherapy regimen for non-Hodgkin's lymphoma. BMC cancer 2024 — PMC11559180.
- Evidences for Mutant Huntingtin Inducing Musculoskeletal and Brain Growth Impairments via Disturbing Testosterone Biosynthesis in Male Huntington Disease Animals. Cells 2022 — PMC9737670.
- The dietary ingredient, genistein, stimulates cathelicidin antimicrobial peptide expression through a novel S1P-dependent mechanism. The Journal of nutritional biochemistry 2014 — PMC4441818.
- PubMed PMID:7529412 — UniProt-cited evidence.
- PubMed PMID:7615076 — UniProt-cited evidence.
- PubMed PMID:7890387 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CAMP (Cathelicidin antimicrobial peptide). Expected localisation Cytoplasmic staining in subsets of immune cells (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A05475 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); 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 A05475); 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-CAMP, 5 μg/mL (datasheet A05475); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Secretion can separate tissue protein and RNA signals (HPA tissue IHC); Isoform / epitope: No isoforms annotated; propeptide is processed (UniProt); Regulation: Staining regulation not reported (UniProt). Evidence: UniProt P49913, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet A05475, PMC11559180. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CAMP/immunohistochemistry-ihc-guide.