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- Table of Contents
CAMP / Cathelicidin antimicrobial peptide · Western blot design guide
Design a Western Blot for CAMP
Real validated CAMP Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CAMP WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
CAMP Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~19.3 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Bone marrow (IHC candidate; verify WB) +2 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Cleaved | |
| Caveat | Proteolytic cleavage | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated CAMP Western Blot Protocols
The A05475 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Human spleen tissue lysate (catalog A05475) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A05475 · 1 μg/mL (catalog A05475) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected CAMP Western Blot Band Size?
CAMP has a predicted 19.3 kDa precursor; signal peptide removal and propeptide cleavage can yield smaller bands, with no empirical band size supplied.
| Band near 19.3 kDa | Unprocessed CAMP precursor is consistent with the predicted mass. |
| Band below 19.3 kDa | Signal peptide removal or propeptide cleavage yields a smaller CAMP product. |
| Little or no band in whole-cell lysate | CAMP may be secreted or stored in neutrophil granules and phagolysosomes. |
| Several bands below 19.3 kDa | Different cleavage states of CAMP may be present. |
💡Expected CAMP appearanceExpect a band near 19.3 kDa for the CAMP precursor, with smaller bands possible after signal peptide removal and propeptide cleavage; the supplied evidence does not establish an empirical band size.
| Predicted CAMP precursor mass | Sets the unprocessed protein reference at 19.3 kDa. |
| Signal peptide at residues 1–30 | Removal makes the processed protein smaller than the precursor. |
| Propeptide at residues 31–131 | Cleavage yields a smaller peptide without a supplied fragment mass. |
| Intrachain disulfides at residues 86–97 and 108–125 | Constrain the propeptide but do not establish a doubled band mass. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CAMP is secreted or stored in neutrophil granules and phagolysosomes. | Check the sampled compartment and whether it contains CAMP-expressing cells. |
| Band higher than expected | The 19.3 kDa precursor may remain uncleaved when a smaller processed product was expected. | Compare precursor and processed samples and check which CAMP region the antibody recognizes. |
| Band lower than expected | Signal peptide removal or propeptide cleavage produces a smaller species. | Check which cleavage state the antibody recognizes. |
| Multiple bands | Precursor and cleaved CAMP products may coexist. | Compare samples with different processing states and verify antibody epitope coverage. |
| Weak or no signal | The detected CAMP region may be absent after cleavage or scarce in the sampled material. | Verify antibody epitope coverage and test a CAMP-containing sample. |
| Fragments below expected size | Propeptide cleavage can release a smaller antimicrobial peptide. | Check sample processing and whether the antibody recognizes the released peptide. |
Experiment evidence
Western blot images showing CAMP band
A05475 · CAMP
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CAMP Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CAMP in Western blot, you can use bone marrow as a high-expression sample.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: CAMP is secreted and stored in neutrophil granules, so lysate signal may depend on the sample preparation.
HPA tissue expression evidence for CAMP
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-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 Western Blot Tips
Deeper troubleshooting and optimisation questions for CAMP, answered from its protein features.
How should CAMP band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could multiple CAMP bands represent isoforms?
Isoforms · Only one isoform is listed. Consider precursor processing before assigning separate bands to isoforms, and check which CAMP region the antibody recognizes.
Are glycosylation or other listed modifications likely to explain a shift?
PTM · No glycosylation sites or modified residues are listed. CAMP does have two disulfide bonds and undergoes signal peptide and propeptide processing. Interpret band differences with those features in mind rather than assigning an unsupported modification.
Does this guide establish induction of CAMP?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for CAMP?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05475 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
What should I quantify when comparing CAMP samples?
Quantitation · Decide whether the measurement concerns the precursor, stored propeptide, or secreted peptide. These can occur in different sample fractions, so quantify the same identified band in the same fraction across samples. The supplied features do not establish an expected apparent band position.
Why might CAMP bands differ from the predicted 19.3 kDa?
Interpretation · The 19.3 kDa prediction is for the precursor. CAMP has a signal peptide at residues 1–30 and a propeptide at residues 31–131, so processing can produce smaller species. Compare bands with the region recognized by your antibody; no empirical apparent band is supplied.
CAMP is stored as a propeptide in neutrophil granules and phagolysosomes and is also secreted, including in sweat. Choose cellular or secreted samples according to the CAMP species you want to detect.
First consider precursor processing and antibody recognition of the signal peptide, propeptide, or remaining peptide. LL-37 can form several oligomers in vitro, but the supplied evidence does not establish that these produce Western blot bands. Avoid assigning an unexpected band to an oligomer without further evidence.
CAMP is stored as a propeptide in neutrophil granules and phagolysosomes and is also secreted, including in sweat. Choose cellular or secreted samples according to the CAMP species you want to detect.
First consider precursor processing and antibody recognition of the signal peptide, propeptide, or remaining peptide. LL-37 can form several oligomers in vitro, but the supplied evidence does not establish that these produce Western blot bands. Avoid assigning an unexpected band to an oligomer without further evidence.
Boster reagents
CAMP Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Cathelicidin CAMP Antibody
Cat # A05475
The catalog reports one anti-CAMP antibody, A05475, with a Western blot image showing Cathelicidin in human spleen tissue lysate at 1 μg/mL. The supplied evidence does not establish performance in other samples or species.
Which to pick: A05475 is the only listed option. Its supplied Western blot image uses human spleen lysate; check suitability for your sample before use.
Source: BosterBio CAMP gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P49913.
- Human Protein Atlas. CAMP tissue expression.
- PMC7646218 — target-verified WB comparison
- PMC5946005 — target-verified WB comparison