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- Table of Contents
CCL19 / C-C motif chemokine 19 · Western blot design guide
Design a Western Blot for CCL19
Real validated CCL19 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CCL19 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
CCL19 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 | ~11 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Cleaved | |
| Caveat | Signal peptide cleavage | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated CCL19 Western Blot Protocols
The M01605-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Human Macrophage Inflammatory Protein 3 beta recombinant protein lysate (catalog M01605-2) |
| 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 | M01605-2; use the WB datasheet starting dilution (standard starting point) |
| 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 CCL19 Western Blot Band Size?
CCL19 has an 11 kDa predicted precursor; secretion and signal-peptide cleavage may affect the detected band, but migration has not been demonstrated.
| Band near 11 kDa | Consistent with the predicted full-length precursor; confirm identity |
| Band below 11 kDa | Could reflect removal of the 1–21 signal peptide; confirm identity |
| Little or no band in whole-cell lysate | Consistent with secretion of CCL19 |
| Band in conditioned medium | Consistent with secreted CCL19; size and identity require confirmation |
💡Expected CCL19 appearanceUniProt predicts an 11 kDa full-length precursor; signal-peptide cleavage may yield a smaller secreted protein, but no empirical band size is supplied, so verify any candidate band with identity controls.
| Full-length precursor mass | Predicted at 11 kDa; observed migration is unreported |
| Signal peptide at residues 1–21 | Cleavage reduces the mass of the secreted protein; its apparent size is unreported |
| Intrachain disulfide between residues 29 and 55 | Adds no mass; any effect of reduction on migration is unestablished |
| Intrachain disulfide between residues 30 and 71 | Adds no mass; any effect of reduction on migration is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CCL19 is secreted | Check conditioned medium alongside lysate |
| Band higher than expected | An uncleaved precursor is possible, but band identity is unconfirmed | Compare lysate and medium and use an independent CCL19 identity control |
| Band lower than expected | Signal-peptide cleavage may yield a smaller protein | Compare with conditioned medium and confirm CCL19 identity |
| Multiple bands | Precursor and signal-cleaved protein are possible; identities are unconfirmed | Compare fractions and verify bands with an independent CCL19 antibody |
| Weak or no signal | Secreted CCL19 may be scarce in the sampled fraction | Check conditioned medium and a CCL19 positive control |
Experiment evidence
Western blot images showing CCL19 band
M01605-2 · CCL19
M01605 · CCL19
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CCL19 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CCL19 in Western blot, you can use appendix tissue.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Because CCL19 is secreted, whole-cell lysates may give weak signal; conditioned medium may be preferable.
HPA tissue expression evidence for CCL19
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 |
|---|---|---|---|---|
| Appendix | germinal center cells | Medium | Protein (IHC) | HPA → |
| Colon | mucosal lymphoid cells | Medium | Protein (IHC) | HPA → |
| Rectum | mucosal lymphoid cells | Medium | Protein (IHC) | HPA → |
| Small intestine | endocrine cells | Medium | Protein (IHC) | HPA → |
| Tonsil | germinal center cells | 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 → |
| 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 CCL19 Western Blot Tips
Deeper troubleshooting and optimisation questions for CCL19, answered from its protein features.
How should CCL19 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 CCL19 bands represent different isoforms?
Isoforms · UniProt lists one isoform and no alternative sequence. Do not assign additional bands to CCL19 isoforms from this record alone; check sample preparation and antibody specificity.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of CCL19?
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.
What transfer method to use for CCL19 Western blot?
Transfer · CCL19 is small: its precursor is predicted at 11 kDa, and its signal peptide spans residues 1–21. Use conditions that retain small proteins, such as a 0.2 µm membrane, and verify transfer near the expected size with a suitable marker.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01605-2 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.
Which samples are useful for quantifying CCL19?
Quantitation · UniProt identifies CCL19 as secreted. Include conditioned medium when measuring released CCL19, and process samples consistently. Specify whether the measurement comes from medium or cell lysate when comparing results.
Why might CCL19 migrate differently from its predicted 11 kDa mass?
Interpretation · The 11 kDa prediction describes the 98-residue precursor. UniProt marks residues 1–21 as a signal peptide, leaving residues 22–98 after processing. Compare the band with a marker near this smaller product, but do not assume processing alone explains its apparent mass; no observed band size was supplied.
UniProt lists two disulfide bonds. Compare reducing and nonreducing samples if mobility differs between preparations. A mobility change is possible, but the feature record alone does not establish that one will occur.
First compare them with the 11 kDa precursor and the product expected after removal of the residues 1–21 signal peptide, using UniProt numbering. The record lists one isoform, two disulfide bonds, and an interaction with TNFAIP6, but none of these features by itself identifies an unexpected band.
UniProt lists two disulfide bonds. Compare reducing and nonreducing samples if mobility differs between preparations. A mobility change is possible, but the feature record alone does not establish that one will occur.
First compare them with the 11 kDa precursor and the product expected after removal of the residues 1–21 signal peptide, using UniProt numbering. The record lists one isoform, two disulfide bonds, and an interaction with TNFAIP6, but none of these features by itself identifies an unexpected band.
Boster reagents
CCL19 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-CCL19 Rabbit Monoclonal Antibody
Cat # M01605-2
Real WB data
Anti-CCL19/Mip 3 Beta Rabbit Monoclonal Antibody
Cat # M01605
Two human-reactive rabbit monoclonal anti-CCL19 antibodies have Western blot images using recombinant protein samples. The supplied evidence does not establish performance with endogenous CCL19 or other species.
Which to pick: Both M01605-2 and M01605 have WB images: M01605-2 uses human MIP-3 beta recombinant protein lysate; M01605 uses CCL19 recombinant protein extracts. Neither image establishes a preference for endogenous samples.
Source: BosterBio CCL19 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99731.
- Human Protein Atlas. CCL19 tissue expression.
- PMC4726905 — target-verified WB comparison
- PMC8517398 — target-verified WB comparison
- PMC7835033 — target-verified WB comparison