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AVP / Vasopressin-neurophysin 2-copeptin · Western blot design guide
Design a Western Blot for AVP
Real validated AVP Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AVP 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
AVP 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 | ~17.3 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Hypothalamus (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 | Glycosylated + Cleaved | |
| Caveat | Processing-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated AVP Western Blot Protocols
The A00384-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A00384-1) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A00384-1) |
| Primary antibody | A00384-1 · 1:1000 (catalog A00384-1) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A00384-1) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A00384-1) |
Section 2
What Is the Expected AVP Western Blot Band Size?
AVP’s predicted precursor is 17.3 kDa; secretion, cleavage, and Asn131 glycosylation may affect detection or migration, but no empirical band size is supplied.
| Band near 17.3 kDa | compatible with the predicted precursor; identity needs confirmation |
| Band below 17.3 kDa | may reflect signal peptide removal or processing at paired basic residues |
| Band above 17.3 kDa | may reflect N-linked glycosylation at Asn131; a visible shift is unproven |
| Several bands at different sizes | may include precursor and processed products; identities need confirmation |
| Little or no band in whole-cell lysate | AVP is secreted |
💡Expected AVP appearanceThe predicted 17.3 kDa mass describes the precursor; signal peptide cleavage, processing, and Asn131 glycosylation may affect migration, but no empirical band size is supplied, so confirm identity with controls.
| Predicted precursor mass | 17.3 kDa is the sequence-based reference, not a validated Western blot band |
| Signal peptide at residues 1–19 | its removal yields a smaller product than the precursor |
| N-linked glycosylation at Asn131 | may alter apparent migration; a visible shift is not established |
| Cleavage on pairs of basic residues | can generate smaller products; their masses are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | AVP is secreted | check conditioned medium alongside lysate |
| Band higher than expected | Asn131 glycosylation may alter migration, but the cause is unconfirmed | compare with an appropriate deglycosylated sample and identity control |
| Band lower than expected | signal peptide removal or cleavage at paired basic residues | check antibody epitope and compare with a precursor control |
| Multiple bands | precursor and processed products may coexist | compare fractions and verify bands with an independent antibody |
| Weak or no signal | secreted AVP may be scarce in the sampled fraction | test conditioned medium and a positive control |
| Fragments below expected size | cleavage at paired basic residues may produce smaller products | check antibody epitope and verify fragment identity with an independent antibody |
Experiment evidence
Western blot images showing AVP band
A00384-1 · AVP
A00384 · AVP
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AVP Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for AVP in Western blot, you can use hypothalamus tissue, where HPA reports high expression.
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 AVP is secreted, whole-tissue lysate may give a weak signal.
HPA tissue expression evidence for AVP
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 → |
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 AVP Western Blot Tips
Deeper troubleshooting and optimisation questions for AVP, answered from its protein features.
How should AVP 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.
Can AVP isoforms explain multiple bands?
Isoforms · UniProt lists one isoform and no alternative sequence. Do not assign multiple bands to isoforms from this record alone. Check which part of the precursor the antibody recognizes and consider processing or antibody specificity when interpreting additional bands.
How should AVP glycosylation be evaluated?
PTM · UniProt lists one N-linked glycosylation site at Asn131, using its precursor numbering. Compare matched untreated and deglycosylated samples if glycosylation is relevant to the band under study. The listed site alone does not establish a detectable mobility shift.
Does this guide establish induction of AVP?
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 AVP?
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 A00384-1 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 AVP species should be quantified?
Quantitation · Define the species recognized by the antibody before quantifying: the 164-residue precursor has a signal peptide at residues 1–19, and UniProt lists cleavage on pairs of basic residues. Quantify the same defined band across matched samples rather than combining bands whose identities are unresolved.
Why might an AVP band differ from 17.3 kDa?
Interpretation · 17.3 kDa is the predicted mass of the 164-residue precursor. UniProt lists a signal peptide at residues 1–19, cleavage on pairs of basic residues, and N-linked glycosylation at Asn131. These features identify possible distinct species, but do not establish a visible band shift or predict an observed band mass.
UniProt lists a glycine amide modification at position 28 in precursor numbering. Consider whether the antibody recognizes a region affected by this modification. Its presence alone does not establish a visible band shift or explain an observed mass difference.
UniProt lists eight disulfide bonds. Compare matched reducing and nonreducing preparations when band behavior is unclear, and record the preparation used. The feature count does not identify which observed band corresponds to a particular disulfide state.
UniProt lists AVP as secreted. Specify whether the blot measures cellular material or a secretion-containing sample, and compare the same sample type across conditions. A band change in one sample type alone does not identify the AVP species responsible.
UniProt lists a glycine amide modification at position 28 in precursor numbering. Consider whether the antibody recognizes a region affected by this modification. Its presence alone does not establish a visible band shift or explain an observed mass difference.
UniProt lists eight disulfide bonds. Compare matched reducing and nonreducing preparations when band behavior is unclear, and record the preparation used. The feature count does not identify which observed band corresponds to a particular disulfide state.
UniProt lists AVP as secreted. Specify whether the blot measures cellular material or a secretion-containing sample, and compare the same sample type across conditions. A band change in one sample type alone does not identify the AVP species responsible.
Boster reagents
AVP 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-AVP Antibody
Cat # A00384-1
Real WB data
Anti-AVP Antibody
Cat # A00384
The catalog reports two anti-AVP antibodies, A00384-1 and A00384, with stated human, mouse, and rat reactivity. Both have Western blot images. The supplied evidence is limited to product captions; no publication evidence is provided.
Which to pick: Both list the same species reactivity and have WB images. Choose A00384 if its documented HEK293T, COS-7, PC12, and CT26 lysates resemble your samples; A00384-1 shows various unnamed cell lines at 1:1000 dilution. The named samples do not establish universal validation.
Source: BosterBio AVP gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P01185.
- Human Protein Atlas. AVP tissue expression.
- PMC7444698 — target-verified WB comparison
- PMC4382529 — target-verified WB comparison
- PMC8447761 — target-verified WB comparison