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PPP1CA / Serine/threonine-protein phosphatase PP1-alpha catalytic subunit · Western blot design guide
Design a Western Blot for PPP1CA
Source-linked PPP1CA Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PPP1CA 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
PPP1CA 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 | ~37.5 kDa | |
| Observed band | ≈38 kDa | |
| Gel | 5–20% (catalog M02801) | |
| Positive control | Kidney (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 | PPP1CB cross-reactivity | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked PPP1CA Western Blot Protocol Options
The M02801 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human Jurkat, human K562, human U87, rat brain, rat C6, mouse brain, mouse NIH/3T3 (catalog M02801) |
| Gel % | 5–20% (catalog M02801) |
| Load | 30 ug; reducing conditions (catalog M02801) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02801) |
| Membrane | nitrocellulose membrane (catalog M02801) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M02801) |
| Primary antibody | M02801 · 1:500 (catalog M02801) |
| Primary incubation | overnight at 4°C (catalog M02801) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M02801) |
| Secondary incubation | 1.5 hour at RT (catalog M02801) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M02801) |
| Detection | ECL (catalog M02801) |
Section 2
What Is the Expected PPP1CA Western Blot Band Size?
PPP1CA is predicted at 37.5 kDa and observed near 38 kDa; the supplied evidence does not establish the cause of the small difference.
| Band near 38 kDa in whole-cell lysate | matches the reported antibody QC band; confirm PPP1CA identity because the antibody also recognizes PPP1CB |
| Several bands at different masses | could include annotated isoforms 1, 2, and 3, whose migration is not supplied |
| Band in a cytoplasmic fraction | consistent with PPP1CA's cytoplasmic location |
| Band in a nuclear fraction | consistent with PPP1CA's nuclear location |
💡Expected PPP1CA appearancePPP1CA's predicted mass is 37.5 kDa, while antibody QC reports a band near 38 kDa; confirm its identity because that antibody also recognizes PPP1CB.
| Predicted mass of 37.5 kDa | provides the sequence-based reference for the band reported near 38 kDa |
| Splice isoform 1 | its individual mass and migration are not supplied |
| Splice isoform 2 | may differ in size from other isoforms; its mass is not supplied |
| Splice isoform 3 | may differ in size from other isoforms; its mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | signal may be below detection despite annotated cytoplasmic and nuclear localization | check a positive-control lysate and inspect cytoplasmic and nuclear fractions |
| Band higher than expected | the supplied features do not establish a higher migrating form | check the size marker and confirm identity with PPP1CA depletion or a second antibody |
| Band lower than expected | an isoform is possible, but isoform masses are not supplied | confirm identity with PPP1CA depletion and compare an independent antibody |
| Multiple bands | isoforms 1, 2, and 3 are annotated, but distinct bands are not established | test band identity with PPP1CA depletion and isoform-specific controls |
| Weak or no signal | PPP1CA may be sparse in the sampled fraction | check a positive control and assess both cytoplasmic and nuclear fractions |
Experiment evidence
Western blot images showing PPP1CA band
M02801 · PPP1CA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PPP1CA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PPP1CA in Western blot, you can use kidney lysate, which has high HPA expression.
Positive control: Kidney (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: An intracellular kidney lysate is suitable, while HPA-undetected adipose tissue provides a candidate negative control.
HPA tissue expression evidence for PPP1CA
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 |
|---|---|---|---|---|
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Rectum | glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PPP1CA Western Blot Tips
Deeper troubleshooting and optimisation questions for PPP1CA, answered from its protein features.
How should PPP1CA 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 might PPP1CA isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Isoform 2 replaces E18 with EGSRVLTPHCAP; isoform 3 lacks residues 19–62. Check whether the antibody recognizes the sequence retained in each isoform before assigning bands. These sequence differences do not establish how the isoforms will migrate.
Which PPP1CA modifications matter when interpreting a blot?
PTM · In UniProt coordinates, residue 2 is listed as N-acetylserine and phosphoserine; S22, Y306, T320 and S325 are phosphorylated sites, and K305 is N6-acetylated. These annotations identify possible site-specific signals but do not establish detectable band shifts.
Match any site-specific antibody’s numbering to the UniProt sequence before comparing signals, especially across isoforms. Isoform 3 lacks canonical residues 19–62, including S22; isoform 2 changes the sequence at E18. Compare each site-specific signal with PPP1CA measured in the same samples.
Match any site-specific antibody’s numbering to the UniProt sequence before comparing signals, especially across isoforms. Isoform 3 lacks canonical residues 19–62, including S22; isoform 2 changes the sequence at E18. Compare each site-specific signal with PPP1CA measured in the same samples.
Does this guide establish induction of PPP1CA?
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 PPP1CA?
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 M02801 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 PPP1CA be quantified across cell fractions?
Quantitation · PPP1CA is listed in the cytoplasm and nucleus, including the nucleoplasm and nucleolus. Quantify matched fractions consistently and account for PPP1CA distribution between them when interpreting changes in a single fraction.
Why does PPP1CA appear near 38 kDa instead of 37.5 kDa?
Interpretation · The observed band is approximately 38 kDa, close to the canonical sequence’s predicted 37.5 kDa. Apparent mass can differ from calculated mass; the listed modifications alone do not establish the cause of this difference.
Check antibody recognition against the three isoform sequences and compare the bands with the approximately 38 kDa observation. UniProt lists no glycosylation sites, signal peptide or propeptide. Its listed modifications and isoforms are candidates to investigate, not proof that they caused an extra band.
Check antibody recognition against the three isoform sequences and compare the bands with the approximately 38 kDa observation. UniProt lists no glycosylation sites, signal peptide or propeptide. Its listed modifications and isoforms are candidates to investigate, not proof that they caused an extra band.
Boster reagents
PPP1CA 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-PPP1CA+PPP1CB Rabbit Monoclonal Antibody
Cat # M02801
M02801 is a rabbit monoclonal antibody against PPP1CA and PPP1CB. Its supplied Western blot image reports an approximately 38 kDa band in human cell, rat cell and brain, and mouse cell and brain lysates. The evidence does not distinguish PPP1CA from PPP1CB.
Which to pick: M02801 is the only listed option. It reports human, mouse, and rat reactivity and includes a Western blot image using samples from all three species. Choose it when a combined PPP1CA/PPP1CB signal is acceptable; the supplied evidence does not establish PPP1CA-specific detection.
Source: BosterBio PPP1CA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P62136.
- Human Protein Atlas. PPP1CA tissue expression.
- PMC11633043 — target-verified WB comparison
- PMC10698990 — target-verified WB comparison
- PMC11569623 — target-verified WB comparison