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- Table of Contents
PABPC5 / Polyadenylate-binding protein 5 · Western blot design guide
Design a Western Blot for PABPC5
Source-linked PABPC5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PABPC5 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
PABPC5 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 | ~43.3 kDa | |
| Observed band | 72 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Fallopian tube (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked PABPC5 Western Blot Protocol Options
The A14533 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | the lysates from HeLa cells (catalog A14533) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A14533; 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 PABPC5 Western Blot Band Size?
PABPC5 is predicted at 43.3 kDa, while antibody QC reports 72 kDa; the cause of this difference is not established.
| Band near 72 kDa | Empirical band reported in cell lysates; PABPC5 identity still needs confirmation. |
| Band near 43.3 kDa | Matches the predicted mass; band identity still needs confirmation. |
| Additional discrete band | Isoforms 1 and 2 could contribute, but distinct migration is unproven. |
| Single band despite two isoforms | Isoforms 1 and 2 are annotated, but separate visible bands are not established. |
💡Expected PABPC5 appearanceUniProt predicts 43.3 kDa, while antibody QC reports a 72 kDa band in cell lysates; its identity and the reason for the difference require controls such as peptide blocking or knockdown.
| UniProt predicted mass | 43.3 kDa is the sequence-based reference, not a measured migration. |
| Isoform 1 | Its individual mass and migration are not supplied. |
| Isoform 2 | Its individual mass and migration are not supplied. |
| Isoforms 1 and 2 | Different forms could affect band size, but their relative sizes and separation are unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoplasmic PABPC5 may be below detection or poorly recovered. | Check lysate loading and extraction; compare with HeLa or K562 lysate. |
| Band higher than expected | The reported 72 kDa band exceeds the 43.3 kDa prediction; the cause is unestablished. | Confirm identity with peptide blocking or PABPC5 knockdown. |
| Band lower than expected | Isoforms are annotated, but no isoform mass or migration is supplied. | Confirm PABPC5 identity and compare isoform-specific evidence. |
| Multiple bands | Isoforms 1 and 2 are possible contributors; their separation is unproven. | Use peptide blocking or knockdown to identify specific bands. |
| Weak or no signal | PABPC5 abundance or assay sensitivity may be insufficient. | Check loading, antibody conditions, and a HeLa or K562 positive-control lysate. |
Experiment evidence
Western blot images showing PABPC5 band
A14533 · PABPC5
A14533 · PABPC5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PABPC5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PABPC5 in Western blot, you can use fallopian tube tissue, the highest-scoring HPA positive sample.
Positive control: Fallopian tube (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: No tissue is reported as not detected, so use siRNA knockdown or a KO line for a clear negative control.
HPA tissue expression evidence for PABPC5
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 |
|---|---|---|---|---|
| Fallopian tube | glandular cells | High | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
Section 3
Advanced PABPC5 Western Blot Tips
Deeper troubleshooting and optimisation questions for PABPC5, answered from its protein features.
How should PABPC5 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.
How could PABPC5 isoforms affect band interpretation?
Isoforms · UniProt lists two isoforms. Isoform 2 lacks residues 1–32 of the canonical sequence, so it has a shorter sequence. That difference alone does not establish where either isoform will migrate on a blot.
An antibody recognizing canonical residues 1–32 may miss isoform 2 because that segment is absent. Check the antibody’s stated epitope against the UniProt coordinates before interpreting an absent isoform band.
An antibody recognizing canonical residues 1–32 may miss isoform 2 because that segment is absent. Check the antibody’s stated epitope against the UniProt coordinates before interpreting an absent isoform band.
Can annotated modifications explain the 72 kDa band?
PTM · The supplied features list no modified residues or glycosylation sites. They therefore provide no annotated modification that explains the 72 kDa apparent band; they also do not rule out unlisted modifications.
Does this guide establish induction of PABPC5?
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 PABPC5?
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 A14533 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 PABPC5 bands be chosen for quantitation?
Quantitation · Define the band to measure consistently across samples. Because two isoforms are listed and the reported 72 kDa band differs from the 43.3 kDa predicted mass, establish which band represents PABPC5 before comparing its intensity.
Why might PABPC5 appear at 72 kDa instead of 43.3 kDa?
Interpretation · The supplied apparent band is 72 kDa, while the predicted mass is 43.3 kDa. The listed features do not establish the cause of that difference. Confirm the band’s identity before assigning it to PABPC5.
Isoform 2 lacks canonical residues 1–32, making it a possible identity to investigate for a lower band. Size alone cannot identify it; check whether the antibody epitope is retained in isoform 2.
Isoform 2 lacks canonical residues 1–32, making it a possible identity to investigate for a lower band. Size alone cannot identify it; check whether the antibody epitope is retained in isoform 2.
Boster reagents
PABPC5 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-PABPC5 Antibody
Cat # A14533
The catalog reports one anti-PABPC5 antibody, A14533, with reported human, mouse, and rat reactivity. Its Western blot image shows HeLa cell lysate; the supplied evidence does not show testing in other samples.
Which to pick: A14533 is the only listed option. Choose it if its reported reactivity fits your sample; the supplied Western blot example uses HeLa cell lysate, so performance in other sample types is not shown.
Source: BosterBio PABPC5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96DU9.
- Human Protein Atlas. PABPC5 tissue expression.
- PMC7183103 — target-verified WB comparison