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- Table of Contents
PVALB / Parvalbumin alpha · Western blot design guide
Design a Western Blot for PVALB
Real validated PVALB Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PVALB 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
PVALB 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 | ~12.1 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PVALB Western Blot Protocols
The M04041 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human cerebellum lysate (catalog M04041) |
| 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 | M04041; 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 PVALB Western Blot Band Size?
PVALB is predicted at 12.1 kDa; its documented acetylation and phosphorylation have no demonstrated effect on Western-blot migration.
| Band near 12.1 kDa | Consistent with PVALB; confirm identity with antibody controls |
| Slightly shifted band near 12.1 kDa | Could reflect a modified PVALB state; migration is unverified |
| Nearby doublet | Could reflect modified and unmodified PVALB; confirm before assigning either band |
| No band near 12.1 kDa | PVALB is not detected in that sample under these conditions |
💡Expected PVALB appearanceUniProt predicts 12.1 kDa for PVALB; no empirical band size or demonstrated migration shift is supplied, so identify a candidate band using a positive control and antibody specificity controls.
| UniProt predicted mass | Places full-length PVALB near 12.1 kDa before any unverified migration effects |
| N-acetylserine at Ser2 | A documented modification with no demonstrated visible band shift |
| Phosphoserine at Ser2 | A documented modification with no demonstrated visible band shift |
| Phosphoserine at Ser24 | A documented modification with no demonstrated visible band shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PVALB may be below detection in the tested lysate | Check a human cerebellum lysate positive control and sample loading |
| Band higher than expected | The supplied features do not establish a higher migrating PVALB species | Compare with a positive control and check antibody specificity |
| Band lower than expected | The supplied features do not establish PVALB cleavage | Check sample integrity and confirm band identity with a positive control |
| Multiple bands | Modification states are possible, but distinct bands are unverified | Compare with a positive control and assess specificity; use phosphatase treatment to investigate phosphorylation |
| Weak or no signal | PVALB abundance or detection may be insufficient | Check loading, antibody performance, and a human cerebellum lysate positive control |
Experiment evidence
Western blot images showing PVALB band
M04041 · PVALB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PVALB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PVALB in Western blot, you can use cerebellum tissue lysate.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA reports high expression in cerebellum and no detected expression in adipose tissue, making tissue controls feasible.
HPA tissue expression evidence for PVALB
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 |
|---|---|---|---|---|
| Cerebellum | molecular layer cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal 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 → |
| 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 PVALB Western Blot Tips
Deeper troubleshooting and optimisation questions for PVALB, answered from its protein features.
How should PVALB 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 a second PVALB band represent a listed isoform?
Isoforms · UniProt supplies one isoform and no alternative sequence. A second band therefore cannot be assigned to a documented PVALB isoform from these features alone. Verify its identity before interpreting it as PVALB.
Which PVALB modifications matter when interpreting bands?
PTM · UniProt lists N-acetylserine and phosphoserine at position 2, and phosphoserine at position 24. These are UniProt coordinates; paper or antibody numbering may differ. The features do not specify when either phosphorylation occurs or whether it changes mobility.
Does this guide establish induction of PVALB?
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 PVALB Western blot?
Transfer · PVALB is a small, 12.1-kDa protein. Use transfer conditions and a membrane that retain proteins of this size, and check the membrane and post-transfer gel if the signal is weak. The supplied features do not specify a particular transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M04041 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 PVALB signal be quantified?
Quantitation · Quantify a consistently identified PVALB band using matched loading normalization. If the goal is phosphorylation, total PVALB signal alone cannot distinguish phosphorylation at UniProt positions 2 and 24 or measure their occupancy.
Should PVALB run exactly at its predicted 12.1 kDa?
Interpretation · Use 12.1 kDa as the predicted mass for the 110-residue protein, not a guaranteed band position. No observed band position is supplied. The listed modifications alone do not establish a visible shift or explain a difference from calculated mass.
Compare them with the predicted 12.1-kDa mass and verify their identity. The supplied features list no glycosylation sites, disulfides, signal peptide, propeptide, or alternative sequence. They also do not establish that the listed acetylation or phosphorylation produces a visible band shift.
Compare them with the predicted 12.1-kDa mass and verify their identity. The supplied features list no glycosylation sites, disulfides, signal peptide, propeptide, or alternative sequence. They also do not establish that the listed acetylation or phosphorylation produces a visible band shift.
Boster reagents
PVALB 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-Parvalbumin PVALB Rabbit Monoclonal Antibody
Cat # M04041
M04041 is a rabbit monoclonal anti-PVALB antibody listed for human, mouse, and rat. Its supplied WB image shows parvalbumin expression in human cerebellum lysate; no mouse or rat WB examples are provided.
Which to pick: M04041 is the only listed option. It has a WB image using human cerebellum lysate; mouse and rat reactivity is listed without corresponding WB examples.
Source: BosterBio PVALB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P20472.
- Human Protein Atlas. PVALB tissue expression.
- PMC4729132 — target-verified WB comparison
- PMC13598087 — target-verified WB comparison
- PMC11088852 — target-verified WB comparison