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- Table of Contents
CAPZB / F-actin-capping protein subunit beta · Western blot design guide
Design a Western Blot for CAPZB
Source-linked CAPZB Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CAPZB 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
CAPZB 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 | ~30.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 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 | 3 isoform(s) |
Section 1
Source-Linked CAPZB Western Blot Protocol Options
The A05623 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 A05623) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A05623) |
| Primary antibody | A05623 · 1:1000 (catalog A05623) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A05623) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A05623) |
Section 2
What Is the Expected CAPZB Western Blot Band Size?
CAPZB is predicted at 30.6 kDa; isoforms and reported modifications may affect migration, but no distinct band size has been demonstrated.
| Band near 30.6 kDa | Consistent with the predicted CAPZB mass; confirm identity with antibody controls. |
| Several discrete bands | Isoforms 1, 2, and 3 may contribute, but distinct migration has not been established. |
| Weak band in a soluble fraction | CAPZB is associated with the cytoskeleton and may be recovered in another fraction. |
| Band with altered mobility near 30.6 kDa | Reported acetylation or phosphorylation could affect migration, but a shift has not been demonstrated. |
💡Expected CAPZB appearanceUniProt predicts 30.6 kDa for CAPZB; no empirical band size is supplied, so use a band near that mass as a starting point and confirm its identity with antibody controls.
| UniProt predicted mass | Places the reference band near 30.6 kDa. |
| Isoform 1 | Its individual mass and migration relative to the other isoforms are not supplied. |
| Isoform 2 | Its sequence variation may affect size, but its relative mass is not supplied. |
| Isoform 3 | Its sequence variation may affect size, but its relative mass is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoskeleton-associated CAPZB may be poorly recovered by the extraction method. | Check the cytoskeletal fraction and verify extraction and antibody performance. |
| Band higher than expected | An isoform or modified CAPZB is possible, but the cause of a shift is unestablished. | Compare isoform expression and confirm band identity with an independent antibody. |
| Band lower than expected | An isoform or protein breakdown is possible; no smaller CAPZB mass is supplied. | Use protease inhibitors and confirm the band with an independent antibody. |
| Multiple bands | CAPZB has three named isoforms, though distinct bands are not established. | Check isoform expression and compare bands using an independent antibody. |
| Weak or no signal | Extraction may recover little cytoskeleton-associated CAPZB. | Compare soluble and cytoskeletal fractions and check a positive control. |
| Fragments below expected size | Protein breakdown during sample handling is possible. | Prepare fresh samples with protease inhibitors and verify fragment identity. |
Experiment evidence
Western blot images showing CAPZB band
A05623 · CAPZB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CAPZB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CAPZB in Western blot, you can use appendix tissue lysate, which HPA scores High.
Positive control: Appendix (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: CAPZB is intracellular, and HPA identifies adipose tissue as a potential negative control.
HPA tissue expression evidence for CAPZB
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | High | Protein (IHC) | HPA → |
| Placenta | decidual 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 → |
| Caudate | neuronal cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced CAPZB Western Blot Tips
Deeper troubleshooting and optimisation questions for CAPZB, answered from its protein features.
Where should the main CAPZB band appear?
Band shift · The predicted mass is 30.6 kDa. No empirical apparent band is supplied, so use 30.6 kDa as a reference rather than an established band position.
Could CAPZB isoforms produce different bands?
Isoforms · Three isoforms are listed. In isoform 2, residues 246–272 are replaced by a different sequence; the supplied features give no sequence change for isoform 3. Consider isoform identity when assessing multiple bands, but these features do not establish their apparent masses.
Which CAPZB modifications matter when interpreting bands?
PTM · UniProt lists N-acetylserine and phosphoserine at position 2, and the supplied annotation ‘N6-acetyllysine Phosphoserine’ at position 235. Keep these UniProt coordinates when comparing site-specific reagents; paper or antibody numbering may differ. The position-235 label is mixed, so verify its chemistry before assigning it.
The listed modifications identify possible molecular differences, but their presence alone does not demonstrate a visible shift or explain a difference from 30.6 kDa. Compare band behavior across samples before assigning a shift to a modification.
The listed modifications identify possible molecular differences, but their presence alone does not demonstrate a visible shift or explain a difference from 30.6 kDa. Compare band behavior across samples before assigning a shift to a modification.
Does this guide establish induction of CAPZB?
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 CAPZB?
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 A05623 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 CAPZB be quantified across samples?
Quantitation · CAPZB is annotated in the cytoplasm and cytoskeleton, including myofibrils and sarcomeres. Keep sample preparation and the measured fraction consistent across samples so differences in recovery are less likely to affect the comparison.
How should unexpected CAPZB bands be interpreted?
Interpretation · Check whether bands could reflect the three listed isoforms, especially the sequence replacement in isoform 2. The supplied features provide no observed band positions, so band size alone cannot identify an isoform or modification; confirm antibody specificity before assigning an unexpected band.
CAPZB is the beta subunit of an alpha–beta F-actin capping complex and is also a dynactin subunit. Those associations do not establish that a higher band is a CAPZB complex on a Western blot. Check sample preparation and band specificity before making that assignment.
CAPZB is the beta subunit of an alpha–beta F-actin capping complex and is also a dynactin subunit. Those associations do not establish that a higher band is a CAPZB complex on a Western blot. Check sample preparation and band specificity before making that assignment.
Boster reagents
CAPZB 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-CAPZB Antibody
Cat # A05623
The catalog reports A05623, an anti-CAPZB antibody with reported human, mouse, and rat reactivity. Its Western blot image uses cell-line extracts at 1:1000 dilution and 25 µg per lane. The caption does not identify the cell lines or provide species-specific results.
Which to pick: A05623 is the only listed CAPZB antibody and has a Western blot image. Check whether its reported reactivity and shown blot conditions fit your sample; the caption does not establish which species were tested in the image.
Source: BosterBio CAPZB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P47756.
- Human Protein Atlas. CAPZB tissue expression.
- PMC8136287 — target-verified WB comparison
- PMC11660498 — target-verified WB comparison
- PMC13333883 — target-verified WB comparison