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PTHLH / Parathyroid hormone-related protein · Western blot design guide
Design a Western Blot for PTHLH
Source-linked PTHLH Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PTHLH 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
PTHLH Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~20.2 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Caudate (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 | Precursor processing | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked PTHLH Western Blot Protocol Options
The A01321 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A01321 · 1:800 (catalog A01321) |
| 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 PTHLH Western Blot Band Size?
PTHLH has a predicted 20.2 kDa precursor; secretion, cleavage, and three isoforms may affect detection or migration, but no specific observed band is established.
| Band near 20.2 kDa | Could represent the predicted full-length PTHLH precursor; confirm its identity. |
| Band below 20.2 kDa | Could reflect signal-peptide or propeptide cleavage; its identity needs confirmation. |
| Little or no band in whole-cell lysate | PTHLH is secreted, so some protein may be found in conditioned medium. |
| Several discrete bands | Could reflect isoforms 1, 2, and 3 or processing; distinct migration is not established. |
💡Expected PTHLH appearanceThe full-length PTHLH precursor is predicted at 20.2 kDa; cleavage may yield smaller species, but no empirical band size or resolvable isoform pattern is supplied, so confirm band identity with appropriate controls.
| Predicted full-length mass | The 177-residue precursor has a calculated mass of 20.2 kDa, not a validated migration position. |
| Signal peptide at residues 1–24 | Its cleavage can make a mature species smaller than the precursor. |
| Propeptide at residues 25–34 | Its cleavage can further reduce the size of a processed species. |
| Splice isoforms 1, 2, and 3 | They may differ in size, but their masses and distinct migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PTHLH can be secreted. | Probe matched conditioned medium and cell lysate. |
| Band lower than expected | Signal-peptide or propeptide cleavage may produce a smaller species. | Compare precursor and processed samples using antibodies to different regions. |
| Multiple bands | Isoforms or proteolytic processing may contribute. | Check band identity with an independent PTHLH antibody and appropriate controls. |
| Weak or no signal | Secretion may reduce the amount retained in cells. | Check conditioned medium and include a positive control. |
| Fragments below expected size | PTHLH has cleavage features, but fragment masses are not supplied. | Compare antibodies to different regions and assess sample handling. |
Experiment evidence
Western blot images showing PTHLH band
A01321 · PTHLH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PTHLH Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PTHLH in Western blot, you can use caudate tissue, which has medium HPA expression.
Positive control: Caudate (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: Because PTHLH is secreted, whole-cell lysates may give weak signal; consider conditioned medium.
HPA tissue expression evidence for PTHLH
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 |
|---|---|---|---|---|
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells - cytoplasm/membrane | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
| Colon | peripheral nerve/ganglion | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PTHLH Western Blot Tips
Deeper troubleshooting and optimisation questions for PTHLH, answered from its protein features.
What should I check when unexpected PTHLH bands appear?
Band shift · Compare their detection with the antibody epitope, the signal peptide and propeptide coordinates, and the C-terminal isoform changes. Check whether the bands occur in medium, cytoplasmic, or nuclear samples. The supplied features suggest possibilities for follow-up, but no observed band sizes are available to identify a band conclusively.
How could PTHLH isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Relative to the canonical sequence, isoform 2 lacks residues 176–177, while isoform 3 replaces those residues with a longer C-terminal sequence. An antibody targeting this region may detect the isoforms differently. These are UniProt precursor coordinates; check the antibody’s numbering convention before comparing epitopes.
Can annotated modifications explain extra PTHLH bands?
PTM · This record lists no glycosylation sites or modified residues. Do not assign an extra band to a specific modification from these features. Consider the annotated precursor regions and isoform differences, then use antibody epitope information to assess which species could be detected.
Does this guide establish induction of PTHLH?
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 PTHLH?
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 A01321 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.
How should I quantify PTHLH across samples?
Quantitation · Quantify a consistently identified band within the same sample fraction and detection setup. Because PTHLH has three isoforms and is annotated in secreted, cytoplasmic, and nuclear locations, state which fraction and band you measured. Do not combine distinct bands as total PTHLH without evidence that the antibody recognizes each species comparably.
Why might PTHLH bands differ from the predicted 20.2 kDa?
Interpretation · 20.2 kDa is the predicted mass of the 177-residue precursor. UniProt annotates a signal peptide at residues 1–24 and a propeptide at 25–34, so a detected product may contain fewer residues. No empirical band size is supplied; these features alone cannot establish a visible shift or explain a particular band.
Check whether the antibody epitope lies in the signal peptide (UniProt residues 1–24), propeptide (25–34), or the remaining sequence. Detection can depend on which region a sample contains. UniProt also flags cleavage on pairs of basic residues but supplies no cleavage coordinates here, so do not assign a band to a specific product from this record alone.
UniProt places PTHLH in secreted, cytoplasmic, and nuclear locations. Choose a sample fraction that matches the species you intend to measure, and compare the same fraction across samples. A difference between medium and cell lysate may reflect distribution as well as abundance.
Check whether the antibody epitope lies in the signal peptide (UniProt residues 1–24), propeptide (25–34), or the remaining sequence. Detection can depend on which region a sample contains. UniProt also flags cleavage on pairs of basic residues but supplies no cleavage coordinates here, so do not assign a band to a specific product from this record alone.
UniProt places PTHLH in secreted, cytoplasmic, and nuclear locations. Choose a sample fraction that matches the species you intend to measure, and compare the same fraction across samples. A difference between medium and cell lysate may reflect distribution as well as abundance.
Boster reagents
PTHLH 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-PTH-rP PTHLH Antibody
Cat # A01321
A01321 is an anti-PTHLH antibody listed for human, mouse, and rat. Its WB image shows HeLa cell analysis at a 1:800 dilution; the supplied evidence does not show WB results in mouse or rat samples.
Which to pick: A01321 is the only listed option. Its HeLa WB image provides a tested starting context at 1:800; confirm performance in your sample, especially for mouse or rat work.
Source: BosterBio PTHLH gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P12272.
- Human Protein Atlas. PTHLH tissue expression.
- PMC3396650 — target-verified WB comparison
- PMC10950337 — target-verified WB comparison
- PMC4393566 — target-verified WB comparison