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- Table of Contents
VIRMA / Protein virilizer homolog · Western blot design guide
Design a Western Blot for VIRMA
Real validated VIRMA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-VIRMA 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
VIRMA 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 | ~202 kDa | |
| Observed band | ~202 kDa | |
| Gel | 5–20% (catalog A32283-1) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Heart muscle (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated VIRMA Western Blot Protocols
The A32283-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human PC-3, human SK-OV-3, human K562 (catalog A32283-1) |
| Gel % | 5–20% (catalog A32283-1) |
| Load | 30 ug; reducing conditions (catalog A32283-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A32283-1) |
| Membrane | nitrocellulose membrane (catalog A32283-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A32283-1) |
| Primary antibody | A32283-1 · 0.5 μg/mL (catalog A32283-1) |
| Primary incubation | overnight at 4°C (catalog A32283-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A32283-1) |
| Secondary incubation | 1.5 hour at RT (catalog A32283-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A32283-1) |
| Detection | ECL (catalog A32283-1) |
Section 2
What Is the Expected VIRMA Western Blot Band Size?
VIRMA is predicted at 202 kDa and observed at ~202 kDa; named isoforms exist, but distinct band sizes are unestablished.
| Band at ~202 kDa | Matches both the predicted mass and the observed VIRMA band |
| Several bands near 202 kDa | Could reflect isoforms 1, 2, 3, and 4; distinct migration is unconfirmed |
| Stronger ~202 kDa band in nuclear fraction | Consistent with VIRMA being mainly nuclear |
| Fainter ~202 kDa band in cytoplasmic fraction | Consistent with a smaller cytoplasmic fraction of VIRMA |
💡Expected VIRMA appearanceVIRMA has a predicted mass of 202 kDa and an empirical band at ~202 kDa; confirm band identity with ordinary controls.
| UniProt predicted mass | Places full-length VIRMA near 202 kDa, matching the observed ~202 kDa band |
| Isoform 1 | Apparent size relative to the other named isoforms is unspecified |
| Isoform 2 | Apparent size relative to the other named isoforms is unspecified |
| Isoforms 3 and 4 | Their apparent sizes and separation from other isoforms are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| Multiple bands | Named splice isoforms are possible, but their migration is unknown | Confirm band identity by VIRMA depletion and compare fractions |
| Weak or no signal | VIRMA is mainly nuclear, so a cytoplasmic fraction may contain little target | Check a nuclear fraction and verify sample loading |
| Band higher than expected | The identity of a band above the observed ~202 kDa position is unestablished | Check molecular-weight markers and test whether VIRMA depletion removes the band |
| Band lower than expected | An isoform or protein breakdown is possible; no lower band size is established | Check sample integrity and test band identity by VIRMA depletion |
| Fragments below expected size | Protein breakdown during sample handling is possible | Prepare fresh samples with protease inhibitors and compare with the ~202 kDa band |
Experiment evidence
Western blot images showing VIRMA band
A32283-1 · VIRMA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for VIRMA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for VIRMA in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Heart muscle (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: VIRMA is mainly nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for VIRMA
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Colon | peripheral nerve/ganglion | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced VIRMA Western Blot Tips
Deeper troubleshooting and optimisation questions for VIRMA, answered from its protein features.
How should VIRMA 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 VIRMA isoforms produce smaller bands?
Isoforms · Yes. Four isoforms are listed. Isoform 2 lacks canonical residues 1378–1812, and isoform 4 lacks 1148–1812; both could migrate below full-length VIRMA. Isoform 3 replaces residues 1684–1812 with a different sequence. Check whether the antibody recognizes a region retained in the isoform under study.
Define whether the measurement targets canonical VIRMA or multiple isoforms. Isoforms 2 and 4 lose substantial C-terminal sequence, while isoform 3 has a replacement C terminus. An antibody against a region missing from an isoform will not quantify that isoform; sum bands only when their identities and antibody recognition are established.
Define whether the measurement targets canonical VIRMA or multiple isoforms. Isoforms 2 and 4 lose substantial C-terminal sequence, while isoform 3 has a replacement C terminus. An antibody against a region missing from an isoform will not quantify that isoform; sum bands only when their identities and antibody recognition are established.
Which phosphorylation sites matter when interpreting VIRMA bands?
PTM · UniProt lists phosphoserine at 133, 138, 173, 222, and 1579; phosphothreonine at 184 and 1708; and phosphotyrosine at 914. These are canonical UniProt coordinates, which may differ from antibody or paper numbering. Their presence does not establish a visible band shift.
Does this guide establish induction of VIRMA?
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 VIRMA Western blot?
Transfer · VIRMA is approximately 202 kDa, so verify that your transfer conditions retain and move proteins of this size onto the membrane. Check the transferred high-mass region with a total-protein stain or marker before interpreting a weak VIRMA signal. The supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A32283-1 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.
How should VIRMA be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
How should the ~202 kDa band compare with predicted VIRMA mass?
Interpretation · The observed band is approximately 202 kDa, matching the supplied predicted mass for canonical VIRMA. Use that position as a starting point for identification; mass agreement alone does not establish band identity.
UniProt lists methylarginine at 1723, alternate asymmetric dimethylarginine or omega-N-methylarginine at 1741, and asymmetric dimethylarginine at 1773, 1775, and 1793. These canonical coordinates lie in regions absent from isoforms 2 and 4. The annotations alone do not predict a resolvable shift.
VIRMA is mainly nuclear, in speckles and nucleoplasm, with some cytoplasmic fraction. Its listed isoforms can differ in size, but the supplied features do not identify every possible band. Compare fractions and antibody epitope location, and verify unexpected bands before assigning them to VIRMA.
UniProt lists methylarginine at 1723, alternate asymmetric dimethylarginine or omega-N-methylarginine at 1741, and asymmetric dimethylarginine at 1773, 1775, and 1793. These canonical coordinates lie in regions absent from isoforms 2 and 4. The annotations alone do not predict a resolvable shift.
VIRMA is mainly nuclear, in speckles and nucleoplasm, with some cytoplasmic fraction. Its listed isoforms can differ in size, but the supplied features do not identify every possible band. Compare fractions and antibody epitope location, and verify unexpected bands before assigning them to VIRMA.
Boster reagents
VIRMA 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-KIAA1429/VIRMA Antibody Picoband®
Cat # A32283-1
The catalog reports one anti-VIRMA antibody for Western blotting, A32283-1, with reported human reactivity. Its product image shows a band near the expected 202 kDa in four human cell lysates. The supplied evidence is limited to this product example.
Which to pick: A32283-1 is the only listed option. Its WB image uses PC-3, SK-OV-3, K562 and HEL whole cell lysates, with antibody at 0.5 μg/mL. Choose it when these reported human conditions fit your experiment.
Source: BosterBio VIRMA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q69YN4.
- Human Protein Atlas. VIRMA tissue expression.
- PMC10198946 — target-verified WB comparison
- PMC11379021 — target-verified WB comparison
- PMC12541586 — target-verified WB comparison