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PPP2R2A / Serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B alpha isoform · Western blot design guide
Design a Western Blot for PPP2R2A
Source-linked PPP2R2A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PPP2R2A 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
PPP2R2A 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 | ~51.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Breast (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 | Acetylated | |
| Caveat | Band size unreported | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked PPP2R2A Western Blot Protocol Options
The A04159 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A04159; 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 PPP2R2A Western Blot Band Size?
PPP2R2A has a predicted mass of 51.7 kDa; its two isoforms could affect migration, but no empirical band size or distinct isoform bands are established.
| Band near 51.7 kDa | consistent with the predicted PPP2R2A mass; confirm identity with controls |
| Band above 51.7 kDa | identity or migration difference is unresolved by the supplied features |
| Band below 51.7 kDa | identity or migration difference is unresolved by the supplied features |
| Several bands at different sizes | isoforms 1 and 2 are possible contributors if they migrate differently |
💡Expected PPP2R2A appearanceUniProt predicts 51.7 kDa, but no empirical band size or isoform migration is supplied; confirm any candidate band with antibody and sample controls.
| UniProt predicted mass | places the reference size at 51.7 kDa |
| Isoform 1 | has no supplied isoform-specific mass or migration |
| Isoform 2 | has no supplied isoform-specific mass or migration |
| Alternative splicing into isoforms 1 and 2 | could affect band position, but a resolvable difference is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PPP2R2A expression or detection in the sample is unverified | check a positive lysate and antibody performance |
| Band higher than expected | the supplied features do not establish a size increase | compare with a validated positive control and check antibody specificity |
| Band lower than expected | isoform migration or band identity is unresolved | compare with a validated positive control and check antibody specificity |
| Multiple bands | isoforms 1 and 2 could contribute, but their migration is unknown | verify band identities with isoform-specific controls if available |
| Weak or no signal | sample expression or antibody detection may be insufficient | check a positive lysate and optimize loading and detection |
Experiment evidence
Western blot images showing PPP2R2A band
A04159 · PPP2R2A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PPP2R2A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PPP2R2A in Western blot, you can use breast tissue, which HPA rates as highly expressed.
Positive control: Breast (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 breast and no detection in adipose tissue, supporting tissue-based controls.
HPA tissue expression evidence for PPP2R2A
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 |
|---|---|---|---|---|
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular 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 → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
Section 3
Advanced PPP2R2A Western Blot Tips
Deeper troubleshooting and optimisation questions for PPP2R2A, answered from its protein features.
How should PPP2R2A 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 do the two PPP2R2A isoforms differ?
Isoforms · Isoform 2 replaces the canonical N-terminal MA at positions 1–2 with MFPKFSLRSMFH. The rest of the listed sequence is shared. Check whether your antibody recognizes this changed N terminus or a shared region; the annotation does not establish whether the isoforms resolve as separate bands.
The annotated sequence difference is at the N terminus: isoform 1 begins MA, while isoform 2 substitutes MFPKFSLRSMFH for those two residues. An antibody directed to that differing region could distinguish them; one directed to the shared sequence may detect both. Confirm the antibody's stated epitope.
The annotated sequence difference is at the N terminus: isoform 1 begins MA, while isoform 2 substitutes MFPKFSLRSMFH for those two residues. An antibody directed to that differing region could distinguish them; one directed to the shared sequence may detect both. Confirm the antibody's stated epitope.
Can N-terminal acetylation explain a second band?
PTM · UniProt annotates N-acetylalanine at canonical position 2. This is a UniProt coordinate; antibody or paper numbering may differ. Isoform 2 replaces the canonical first two residues, so do not assume this annotation applies to it. The listed modification alone does not demonstrate a visible band shift.
Does this guide establish induction of PPP2R2A?
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 PPP2R2A?
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 A04159 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 PPP2R2A bands be quantified?
Quantitation · Choose a consistent band or band set and report which you quantified. Because UniProt lists two isoforms, an antibody recognizing shared sequence may measure both if they overlap. The supplied features do not establish their relative abundance or whether they separate on the gel.
Why might PPP2R2A migrate away from its predicted mass?
Interpretation · Its predicted mass is 51.7 kDa. UniProt lists two isoforms and N-acetylalanine at canonical position 2, but these features alone do not establish a visible shift or explain a difference from 51.7 kDa. No observed band position is supplied.
Compare bands with the 51.7 kDa predicted mass and consider the annotated N-terminal isoform difference. PPP2R2A is also described as associating with PP2A core subunits, but that description does not identify an extra band. The supplied features cannot assign an unexpected band without additional experimental evidence.
Compare bands with the 51.7 kDa predicted mass and consider the annotated N-terminal isoform difference. PPP2R2A is also described as associating with PP2A core subunits, but that description does not identify an extra band. The supplied features cannot assign an unexpected band without additional experimental evidence.
Boster reagents
PPP2R2A 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-PPP2R2A Antibody (N-term)
Cat # A04159
the supplier A04159 is an anti-PPP2R2A N-terminal antibody listed for human samples. Its Western blot image shows detection in ZR-75-1 cell lysate at 35 µg per lane. The supplied evidence does not establish performance across other samples or conditions.
Which to pick: A04159 is the only listed option. Choose it when its stated human reactivity and documented ZR-75-1 Western blot conditions fit your experiment.
Source: BosterBio PPP2R2A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P63151.
- Human Protein Atlas. PPP2R2A tissue expression.
- PMC6620383 — target-verified WB comparison
- PMC8455141 — target-verified WB comparison
- PMC4310454 — target-verified WB comparison