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- Table of Contents
PIK3C3 / Phosphatidylinositol 3-kinase catalytic subunit type 3 · Western blot design guide
Design a Western Blot for PIK3C3
Real validated PIK3C3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PIK3C3 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
PIK3C3 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 | ~101.5 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (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 + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated PIK3C3 Western Blot Protocols
The A02356 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse small intestine tissue lysate (catalog A02356) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02356 · 1 μg/ml (catalog A02356) |
| 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 PIK3C3 Western Blot Band Size?
PIK3C3 is predicted at 101.5 kDa; phosphorylation could affect mobility, but no empirical band or demonstrated migration shift is supplied.
| Band near 101.5 kDa | Consistent with the predicted PIK3C3 size; confirm identity with controls |
| Close doublet near 101.5 kDa | Could reflect different phosphorylation states; separation is not established |
| Band slightly displaced from 101.5 kDa | Phosphorylation may affect mobility, but the cause requires testing |
| Single band without a visible shift | Annotated phosphorylation sites need not produce resolvable bands |
💡Expected PIK3C3 appearancePIK3C3 has a predicted mass of 101.5 kDa, but no empirical band size is supplied; assess a band near that size with ordinary identity controls because phosphorylation does not establish a migration shift.
| UniProt predicted mass | Places the reference band near 101.5 kDa |
| AMPK phosphorylation at Thr163 | May affect mobility; no shift is demonstrated |
| AMPK phosphorylation at Ser165 | May affect mobility; no shift is demonstrated |
| Phosphorylation at Ser244 | May affect mobility; no shift is demonstrated |
| Phosphorylation at Ser261 and Ser282 | May affect mobility; no shift is demonstrated |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PIK3C3 occurs at endosomes and autophagosomes; recovery from the preparation may be poor | Check extraction and loading with a positive control |
| Band higher than expected | Phosphorylation is possible, but its effect on migration is unproven | Compare phosphatase-treated material and confirm band identity |
| Band lower than expected | A smaller PIK3C3 product is not established by the supplied features | Check antibody specificity and sample integrity |
| Multiple bands | Different phosphorylation states are possible, but distinct bands are unproven | Compare phosphatase-treated samples and use an independent PIK3C3 antibody |
| Weak or no signal | Recovery of endosomal or autophagosomal PIK3C3 may be limited | Check extraction, loading, and a positive control |
Experiment evidence
Western blot images showing PIK3C3 band
A02356 · PIK3C3
A02356-3 · PIK3C3
M02356 · PIK3C3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PIK3C3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PIK3C3 in Western blot, you can use adrenal gland tissue, which HPA rates as high expression.
Positive control: Adrenal gland (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: PIK3C3 is intracellular, and HPA identifies adipose tissue as a not-detected comparator.
HPA tissue expression evidence for PIK3C3
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | 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 → |
| Caudate | glial cells | Low | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Low | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced PIK3C3 Western Blot Tips
Deeper troubleshooting and optimisation questions for PIK3C3, answered from its protein features.
What band size should I expect for PIK3C3?
Band shift · The predicted mass is 101.5 kDa. No observed band size is supplied, so use this as a reference rather than an exact migration position. The listed phosphorylation sites alone do not establish a visible shift or explain a difference from the predicted mass.
Could PIK3C3 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning additional bands to annotated isoforms.
Which PIK3C3 phosphorylation sites could affect blot interpretation?
PTM · UniProt lists Thr163 and Ser165 phosphorylated by AMPK, plus phosphoserines at 244, 261, and 282. These are UniProt coordinates; antibody or paper numbering may differ. A site-specific signal can change without a detectable shift in total PIK3C3 migration.
Does this guide establish induction of PIK3C3?
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 PIK3C3 Western blot?
Transfer · Choose transfer conditions that retain and recover a protein near 101.5 kDa. Check the membrane and post-transfer gel to verify recovery before interpreting a weak or absent band. The supplied features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02356 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 I quantify PIK3C3 across samples?
Quantitation · Quantify the band near the predicted 101.5 kDa position within a consistent sample fraction and normalize for loading. PIK3C3 is reported at autophagosomes, late endosomes, and the midbody, so changes in fraction recovery can affect comparisons.
How should I investigate unexpected PIK3C3 bands?
Interpretation · Compare bands with the 101.5 kDa prediction and check antibody specificity. The record lists one isoform and five phosphorylation sites, but neither establishes the identity or size of an extra band. Avoid assigning bands to an isoform or modification without additional evidence.
Boster reagents
PIK3C3 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-PIK3C3 Antibody
Cat # A02356
Real WB data
Anti-VPS34/PIK3C3 Antibody Picoband®
Cat # A02356-3
Real WB data
Anti-PI 3 Kinase Class 3 PIK3C3 Rabbit Monoclonal Antibody
Cat # M02356
Three the supplier anti-PIK3C3 antibodies report human, mouse, and rat reactivity and have Western blot images. Captions show mouse small intestine lysate for A02356, human cell lysates for A02356-3, and 293T lysate for M02356. No rat blot is supplied.
Which to pick: For mouse small intestine, consider A02356; for human cell lysates, A02356-3 has the most detailed blot conditions and a reported 102 kDa band. M02356 shows a 293T lysate blot. All three list rat reactivity, but none shows a rat sample.
Source: BosterBio PIK3C3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8NEB9.
- Human Protein Atlas. PIK3C3 tissue expression.
- PMC7111147 — target-verified WB comparison
- PMC11784013 — target-verified WB comparison