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- Table of Contents
PTH / Parathyroid hormone · Western blot design guide
Design a Western Blot for PTH
Real validated PTH Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PTH 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
PTH 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 | ~12.9 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Parathyroid gland (IHC candidate; verify WB) | |
| 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 C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PTH Western Blot Protocols
The M00208-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Parathyroid Hormone recombinant protein (catalog M00208-2) |
| 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 | M00208-2; 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 PTH Western Blot Band Size?
The PTH precursor is predicted at 12.9 kDa; annotated cleavage may yield smaller PTH, but no empirical migration is supplied.
| Band near 12.9 kDa | May represent the full-length PTH precursor; confirm its identity. |
| Band below 12.9 kDa | May represent PTH after signal-peptide and propeptide cleavage. |
| Bands near and below 12.9 kDa | Could reflect precursor and processed PTH; confirm both identities. |
| Little or no band in whole-cell lysate | PTH is secreted. |
💡Expected PTH appearanceUniProt predicts 12.9 kDa for the PTH precursor; signal-peptide and propeptide cleavage may yield a smaller band, but no empirical band size is supplied, so confirm band identity with appropriate controls.
| UniProt-predicted precursor mass | Places the full-length 115-residue PTH precursor near 12.9 kDa. |
| Signal peptide at residues 1–25 | Its cleavage makes PTH smaller than the full-length precursor. |
| Propeptide at residues 26–31 | Its cleavage further reduces the size of processed PTH. |
| Combined precursor processing | Removal of both annotated segments yields a smaller mature product; its apparent mass is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PTH is secreted. | Check conditioned medium or another relevant secreted fraction. |
| Band higher than expected | The band may contain uncleaved PTH precursor. | Compare precursor and secreted fractions and verify band identity. |
| Band lower than expected | Signal-peptide and propeptide cleavage may reduce PTH size. | Check whether the antibody recognizes processed PTH and verify the band. |
| Multiple bands | Precursor and processed PTH may coexist. | Compare cellular and secreted fractions and confirm each band’s identity. |
| Weak or no signal | Secreted PTH may be scarce in the tested fraction. | Check sample fraction, loading, and a suitable positive control. |
Experiment evidence
Western blot images showing PTH band
M00208-2 · PTH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PTH Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PTH in Western blot, you can use parathyroid gland tissue.
Positive control: Parathyroid gland (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: PTH is secreted, so tissue lysates may give weak signal; consider testing the secreted fraction.
HPA tissue expression evidence for PTH
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 |
|---|---|---|---|---|
| Parathyroid gland | glandular cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PTH Western Blot Tips
Deeper troubleshooting and optimisation questions for PTH, answered from its protein features.
How should PTH 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.
Do annotated PTH isoforms explain multiple bands?
Isoforms · UniProt lists one isoform and no alternative sequence. Its annotated signal peptide and propeptide offer a processing-related possibility to investigate, but the supplied features do not identify the cause of any extra band.
Are annotated PTH modifications likely to explain a shifted band?
PTM · The supplied UniProt record lists no glycosylation sites, modified residues, or disulfides. It therefore provides no specific modification to test as the cause of a shift. Compare any observed bands with the precursor and processed regions before assigning an explanation.
Does this guide establish induction of PTH?
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 PTH Western blot?
Transfer · PTH is annotated as a 115-residue precursor with a shorter processed hormone comprising residues 32..115. Select and validate a transfer setup that retains small proteins; check retention with a suitable small-protein control. The UniProt features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M00208-2 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 sample fraction is relevant when quantifying PTH?
Quantitation · UniProt identifies PTH as secreted. For secreted PTH, collect the extracellular fraction and keep collection volume and cell input consistent across samples. State whether the assay targets processed PTH or the 115-residue precursor, since the annotated signal peptide and propeptide distinguish them.
Why might PTH migrate differently from its predicted 12.9 kDa mass?
Interpretation · The 12.9 kDa prediction describes the 115-residue precursor. UniProt annotates a signal peptide at residues 1..25 and a propeptide at 26..31, leaving residues 32..115 in the processed hormone. These features suggest different molecular sizes, but no observed band is supplied, and cleavage alone does not establish a particular apparent mass.
Choose an antibody whose epitope lies within residues 32..115 if the goal is to detect processed PTH. An epitope restricted to the annotated signal peptide (1..25) or propeptide (26..31) would target precursor regions removed during processing. Confirm the vendor's residue numbering against UniProt precursor coordinates.
Check whether the antibody recognizes residues retained in processed PTH (32..115) or the precursor regions (1..31), using UniProt precursor numbering. The record lists one isoform and no modification sites, while the apparent band position is unavailable. These features alone cannot assign an unexpected band to a specific PTH form.
Choose an antibody whose epitope lies within residues 32..115 if the goal is to detect processed PTH. An epitope restricted to the annotated signal peptide (1..25) or propeptide (26..31) would target precursor regions removed during processing. Confirm the vendor's residue numbering against UniProt precursor coordinates.
Check whether the antibody recognizes residues retained in processed PTH (32..115) or the precursor regions (1..31), using UniProt precursor numbering. The record lists one isoform and no modification sites, while the apparent band position is unavailable. These features alone cannot assign an unexpected band to a specific PTH form.
Boster reagents
PTH 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-Parathyroid Hormone Rabbit Monoclonal Antibody
Cat # M00208-2
M00208-2 is a rabbit monoclonal anti-PTH antibody listed as human-reactive. Its Western blot image shows human recombinant PTH; the supplied evidence does not establish performance in endogenous samples.
Which to pick: M00208-2 is the only listed antibody. Choose it if human reactivity and a Western blot image using recombinant PTH fit your design; no comparison with other antibodies is supplied.
Source: BosterBio PTH gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P01270.
- Human Protein Atlas. PTH tissue expression.
- PMC11952747 — target-verified WB comparison