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- Table of Contents
OXT / Oxytocin-neurophysin 1 · Western blot design guide
Design a Western Blot for OXT
Source-linked OXT Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-OXT 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
OXT 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 | ~12.7 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Hypothalamus (IHC candidate; verify WB) +1 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 | 1 isoform(s) |
Section 1
Source-Linked OXT Western Blot Protocol Options
The A01545 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A549 (20ug), SGC7901 (40ug), 3T3-L1 (40ug), H9C2 (40ug) (catalog A01545) |
| 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 | A01545; 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 OXT Western Blot Band Size?
OXT has a predicted 12.7 kDa precursor; secretion and processing may affect the detected band, but no empirical migration is supplied.
| Band near 12.7 kDa | May represent the predicted full-length precursor; confirm its identity with controls |
| Band below 12.7 kDa | May reflect signal-peptide removal or cleavage at paired basic residues |
| Little or no band in whole-cell lysate | OXT is secreted |
| Several bands below 12.7 kDa | May represent products of the annotated cleavage at paired basic residues; confirm their identities |
💡Expected OXT appearanceUniProt predicts a 12.7 kDa precursor, but signal-peptide removal and annotated cleavage may yield smaller products; no empirical band size is supplied, so confirm band identity with appropriate controls.
| Predicted full-length mass | The 125-residue precursor has a calculated mass of 12.7 kDa |
| Signal peptide at residues 1–19 | Its removal yields a smaller product than the precursor |
| Cleavage at paired basic residues | Processing may yield smaller products; their masses are not supplied |
| Gly28 amidation | This modification is annotated, but a visible migration effect is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | OXT is secreted | Check conditioned medium alongside lysate |
| Band lower than expected | Signal-peptide removal or cleavage at paired basic residues may yield a smaller product | Compare with a verified precursor or processed-product control |
| Multiple bands | Annotated processing may produce distinct products, but their migration is unverified | Use a verified positive control to identify the recognized species |
| Weak or no signal | A secreted target may be scarce in the sampled lysate | Test conditioned medium and a positive control |
| Fragments below expected size | Cleavage at paired basic residues may generate smaller products | Confirm fragment identity with an appropriate epitope or processing control |
Experiment evidence
Western blot images showing OXT band
A01545 · OXT
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for OXT Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for OXT in Western blot, you can use hypothalamus tissue.
Positive control: Hypothalamus (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 OXT is secreted, signal in tissue lysate may be low; consider an extracellular sample.
HPA tissue expression evidence for OXT
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 |
|---|---|---|---|---|
| Hypothalamus | neuronal cells | High | Protein (IHC) | HPA → |
| Ovary | follicle 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 | lymphoid tissue | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced OXT Western Blot Tips
Deeper troubleshooting and optimisation questions for OXT, answered from its protein features.
How should OXT 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.
Does OXT have alternative isoforms that explain extra bands?
Isoforms · Only one isoform is listed, with no alternative sequence. Do not assign extra bands to a documented OXT isoform on this evidence.
Is OXT glycosylated?
PTM · No glycosylation sites are listed for OXT. Its supplied features therefore provide no basis for attributing a band shift to glycosylation.
Does this guide establish induction of OXT?
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 OXT?
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 A01545 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 relevant when quantifying OXT?
Quantitation · OXT is annotated as secreted. If measuring production or release, consider the sample medium as well as the cell or tissue fraction, and report which fraction was measured. A signal in one fraction alone does not quantify total OXT.
Should OXT migrate at its predicted 12.7 kDa?
Interpretation · The 12.7 kDa prediction refers to the 125-residue sequence. OXT has a signal peptide at UniProt positions 1–19 and a cleavage-on-paired-basic-residues annotation, so processed species may differ in size. No observed band is supplied; these features alone cannot predict its apparent migration.
The sequence has a signal peptide at UniProt positions 1–19, and OXT is annotated for cleavage on pairs of basic residues. A smaller band could represent a processed species, but band size alone cannot establish its identity. Check which part of OXT the antibody recognizes.
The listed modification is glycine amide at UniProt position 28. Keep that coordinate convention when comparing antibody or paper numbering. The annotation establishes a modification site, but does not establish a visible band shift.
OXT has eight annotated disulfide bonds. Comparing reducing and nonreducing samples may help assess whether disulfide-dependent structure affects migration. The annotation alone does not predict a particular band position or shift.
The sequence has a signal peptide at UniProt positions 1–19, and OXT is annotated for cleavage on pairs of basic residues. A smaller band could represent a processed species, but band size alone cannot establish its identity. Check which part of OXT the antibody recognizes.
The listed modification is glycine amide at UniProt position 28. Keep that coordinate convention when comparing antibody or paper numbering. The annotation establishes a modification site, but does not establish a visible band shift.
OXT has eight annotated disulfide bonds. Comparing reducing and nonreducing samples may help assess whether disulfide-dependent structure affects migration. The annotation alone does not predict a particular band position or shift.
Boster reagents
OXT 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-Neurophysin I (K40) OXT Antibody
Cat # A01545
The catalog reports one anti-OXT antibody, A01545, with stated Human, Mouse, and Rat reactivity. Its WB image caption reports testing at 1:500 on A549, SGC7901, 3T3-L1, and H9C2 whole-cell lysates. No independent publication evidence is supplied.
Which to pick: A01545 is the only listed option. Its WB image uses A549, SGC7901, 3T3-L1, and H9C2 lysates at 1:500; consider whether those tested samples and conditions fit your experiment.
Source: BosterBio OXT gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P01178.
- Human Protein Atlas. OXT tissue expression.
- PMC10030782 — target-verified WB comparison
- PMC9392805 — target-verified WB comparison