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- Table of Contents
PABPN1 / Polyadenylate-binding protein 2 · Western blot design guide
Design a Western Blot for PABPN1
Real validated PABPN1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PABPN1 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
PABPN1 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 | ~32.7 kDa | |
| Observed band | ~50 kDa | |
| Gel | 10% (catalog A02445-2) | |
| Positive control | Appendix (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 | Phosphorylated + Acetylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated PABPN1 Western Blot Protocols
The A02445-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human RT4, human THP-1 (catalog A02445-2) |
| Gel % | 10% (catalog A02445-2) |
| Load | 30 ug; reducing conditions (catalog A02445-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02445-2) |
| Membrane | nitrocellulose membrane (catalog A02445-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02445-2) |
| Primary antibody | A02445-2 · 0.5 μg/mL (catalog A02445-2) |
| Primary incubation | overnight at 4°C (catalog A02445-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02445-2) |
| Secondary incubation | 1.5 hour at RT (catalog A02445-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02445-2) |
| Detection | ECL (catalog A02445-2) |
Section 2
What Is the Expected PABPN1 Western Blot Band Size?
PABPN1 predicts at 32.7 kDa but is observed at ~50 kDa in reducing Western blots; the cause of this difference is not established.
| Band at ~50 kDa | Empirical PABPN1 band in reducing whole-cell lysates; identity still requires controls |
| Band near 33 kDa | Near the 32.7 kDa sequence-predicted mass; identity requires confirmation |
| Several bands at different positions | Could include splice isoforms 1, 2, and 3; their migration is unestablished |
| Additional higher-mass band | Could reflect retained PABPN1 homooligomers; confirm before assigning identity |
💡Expected PABPN1 appearancePABPN1 has a predicted mass of 32.7 kDa but an empirical band at ~50 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm identity with appropriate controls.
| UniProt predicted mass | 32.7 kDa from sequence; the empirical band is ~50 kDa |
| Splice isoform 1 | Its apparent size relative to the other isoforms is not supplied |
| Splice isoform 2 | Its apparent size relative to the other isoforms is not supplied |
| Splice isoform 3 | Its apparent size relative to the other isoforms is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear PABPN1 may be poorly recovered during extraction | Check nuclear extraction and include a known positive lysate |
| Band higher than expected | The observed ~50 kDa band exceeds the 32.7 kDa prediction for an unestablished reason | Compare with the antibody QC band and verify identity by PABPN1 depletion |
| Band lower than expected | A splice isoform or nonspecific band is possible; isoform migration is unknown | Verify with PABPN1 depletion and an independent antibody |
| Multiple bands | PABPN1 has three splice isoforms, but distinct bands are not established | Check which bands respond to PABPN1 depletion |
| Weak or no signal | Nuclear PABPN1 may be underrepresented in the prepared sample | Assess extraction with a nuclear marker and use a positive control |
Experiment evidence
Western blot images showing PABPN1 band
A02445-2 · PABPN1
M02445 · PABPN1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PABPN1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PABPN1 in Western blot, you can use appendix tissue, which shows high HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No tissue is listed as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for PABPN1
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 → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | 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 → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced PABPN1 Western Blot Tips
Deeper troubleshooting and optimisation questions for PABPN1, answered from its protein features.
How should PABPN1 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 PABPN1 isoforms produce different bands?
Isoforms · Three isoforms are listed. Relative to the 306-residue canonical sequence, isoform 2 has GR replaced by SG at residues 295–296 and lacks residues 297–306. This could change its calculated mass, but the features do not establish a resolvable band difference. No sequence change is supplied for isoform 3.
Which PABPN1 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at canonical positions 19, 52, 95, 150, and 235. These sites support considering phosphorylation when investigating band patterns, but they do not establish a visible shift. Keep the UniProt numbering explicit when comparing site-specific antibodies or reports.
Does this guide establish induction of PABPN1?
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 PABPN1 Western blot?
Transfer · The supplied features do not specify a transfer method. Optimize transfer and check retention around both the 32.7 kDa predicted mass and the reported apparent band near 50 kDa; neither value alone establishes the best transfer conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02445-2 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 PABPN1 bands be quantified across samples?
Quantitation · Define the band or band region consistently across samples, using the reported approximately 50 kDa band as a starting reference. If additional bands appear, verify their identity before combining their signals. The three listed isoforms and modification sites alone do not identify which bands should be included.
Why does PABPN1 appear near 50 kDa instead of 32.7 kDa?
Interpretation · 32.7 kDa is the predicted mass, while approximately 50 kDa is the reported apparent band. The listed modifications and isoforms do not establish the cause of that difference. Use the observed position as a reference and verify band identity independently.
Methylation is listed at canonical position 17 and multiple positions from 238 to 298. Positions 238, 259, and 263 have alternate mono- and asymmetric dimethylarginine annotations. These annotations do not show which forms coexist or whether they separate on a blot. Check numbering against the isoform being measured, especially near isoform 2’s altered C terminus.
Consider the listed isoforms and modifications as candidates to investigate, without assigning an unexpected band to either from size alone. PABPN1 is described as a monomer and homooligomer that oligomerizes when bound to poly(A). Verify band identity and compare sample preparation before interpreting additional bands.
Methylation is listed at canonical position 17 and multiple positions from 238 to 298. Positions 238, 259, and 263 have alternate mono- and asymmetric dimethylarginine annotations. These annotations do not show which forms coexist or whether they separate on a blot. Check numbering against the isoform being measured, especially near isoform 2’s altered C terminus.
Consider the listed isoforms and modifications as candidates to investigate, without assigning an unexpected band to either from size alone. PABPN1 is described as a monomer and homooligomer that oligomerizes when bound to poly(A). Verify band identity and compare sample preparation before interpreting additional bands.
Boster reagents
PABPN1 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-PABPN1 Antibody Picoband®
Cat # A02445-2
Real WB data
Anti-PABPN1 Rabbit Monoclonal Antibody
Cat # M02445
The catalog reports two anti-PABPN1 antibodies with Western blot images from human, rat, and mouse lysates. Both captions report a band near 50 kDa, above their stated expected sizes of 33 or 35 kDa; the supplied evidence does not resolve this discrepancy.
Which to pick: Both A02445-2 (polyclonal) and M02445 (monoclonal) have WB images. Choose by antibody format and the closest tested lysate: A02445-2 includes rat and mouse brain samples, while M02445 includes rat PC-12 and mouse Raw264.7 cells.
Source: BosterBio PABPN1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q86U42.
- Human Protein Atlas. PABPN1 tissue expression.
- PMC4859507 — target-verified WB comparison
- PMC3879409 — target-verified WB comparison