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- Table of Contents
PIK3CD / Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit delta isoform · Western blot design guide
Design a Western Blot for PIK3CD
Real validated PIK3CD Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PIK3CD 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
PIK3CD 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 | ~119.5 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated PIK3CD Western Blot Protocols
The M02269 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | (1) K562 cell lysate; (2) RAW 264 (catalog M02269) |
| 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 | M02269; 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 PIK3CD Western Blot Band Size?
PIK3CD is predicted at 119.5 kDa; isoforms and phosphorylation are annotated, but distinct migration has not been demonstrated here.
| Single band near 119.5 kDa | Consistent with the predicted PIK3CD mass; confirm identity with appropriate controls. |
| Two bands at different positions | Could reflect isoforms 1 and 2 if both are expressed and resolve separately. |
| Doublet near 119.5 kDa | Could reflect distinct phosphorylation states; validate with phosphatase treatment. |
| Band displaced from 119.5 kDa | Apparent migration differs from the prediction; identity requires confirmation. |
💡Expected PIK3CD appearancePIK3CD has a predicted mass of 119.5 kDa, but no empirical band size is supplied; use antibody specificity and PIK3CD depletion controls to identify any band.
| Predicted PIK3CD mass | Places the expected full-length band near 119.5 kDa. |
| Alternative splicing | May produce proteins with different apparent sizes; their separation is unknown. |
| Isoform 1 | Its individual mass and migration are not supplied. |
| Isoform 2 | Its individual mass and migration are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | Its identity or apparent migration is uncertain. | Compare with PIK3CD depletion and a second antibody. |
| Band lower than expected | An isoform or degradation product is possible. | Compare antibodies against different epitopes and check sample integrity. |
| Multiple bands | Isoforms 1 and 2 or nonspecific binding may contribute. | Use PIK3CD depletion to identify specific bands. |
| Weak or no signal | PIK3CD abundance or antibody sensitivity may be insufficient. | Check lysate loading and include a known positive lysate. |
| Fragments below expected size | Protein degradation is possible. | Prepare fresh lysate with protease inhibitors and compare epitopes. |
Experiment evidence
Western blot images showing PIK3CD band
M02269 · PIK3CD
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PIK3CD Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PIK3CD in Western blot, you can use appendix tissue lysate, which has medium HPA expression.
Positive control: Appendix (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: Appendix provides a feasible cytoplasmic positive control, though its HPA signal is only medium.
HPA tissue expression evidence for PIK3CD
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 | lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Testis | Leydig cells | Medium | Protein (IHC) | HPA → |
| Tonsil | squamous 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PIK3CD Western Blot Tips
Deeper troubleshooting and optimisation questions for PIK3CD, answered from its protein features.
How should PIK3CD 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 the two PIK3CD isoforms affect band interpretation?
Isoforms · Isoform 2 replaces canonical residues 201–300 with an alternative sequence and lacks residues 302–1044. It therefore differs substantially from full-length isoform 1. Check whether the antibody recognizes a region retained in the isoform before assigning a shorter band.
No. The listed sites, positions 524 and 1039 in the canonical UniProt sequence, fall within residues 302–1044, which are missing in isoform 2. State the numbering convention when comparing these sites with antibody or paper coordinates.
No. The listed sites, positions 524 and 1039 in the canonical UniProt sequence, fall within residues 302–1044, which are missing in isoform 2. State the numbering convention when comparing these sites with antibody or paper coordinates.
Could phosphorylation explain a PIK3CD band shift?
PTM · UniProt lists phosphotyrosine at position 524 and phosphoserine at position 1039, the latter by autocatalysis. These are canonical-sequence coordinates. Their presence alone does not establish a visible shift or explain any difference between apparent and predicted mass.
Does this guide establish induction of PIK3CD?
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 PIK3CD?
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 M02269 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 PIK3CD bands be quantified?
Quantitation · Choose the band corresponding to the isoform being measured and use that assignment consistently across samples. Because isoform 2 lacks canonical residues 302–1044, an antibody recognizing that region would not measure isoform 2. Do not combine bands without establishing what each represents.
Where should full-length PIK3CD appear?
Interpretation · Canonical PIK3CD has a predicted mass of 119.5 kDa. Use this as a reference when identifying the full-length band; no observed band position was supplied, so its apparent migration cannot be specified.
Isoform 2 is a plausible candidate because it lacks canonical residues 302–1044, but size alone cannot identify it. Check the antibody’s recognition region: a region within those missing residues cannot detect isoform 2. The supplied features do not establish an observed band position.
No. PIK3CD forms a heterodimer with a p85 regulatory subunit, PIK3R1, PIK3R2 or PIK3R3. Complex membership alone does not identify a detected band as PIK3CD; use PIK3CD-specific recognition and the isoform sequence information when assigning bands.
Isoform 2 is a plausible candidate because it lacks canonical residues 302–1044, but size alone cannot identify it. Check the antibody’s recognition region: a region within those missing residues cannot detect isoform 2. The supplied features do not establish an observed band position.
No. PIK3CD forms a heterodimer with a p85 regulatory subunit, PIK3R1, PIK3R2 or PIK3R3. Complex membership alone does not identify a detected band as PIK3CD; use PIK3CD-specific recognition and the isoform sequence information when assigning bands.
Boster reagents
PIK3CD 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-PI 3 Kinase p110 delta PIK3CD Rabbit Monoclonal Antibody
Cat # M02269
M02269 is a rabbit monoclonal anti-PIK3CD antibody listed for Western blotting with human, mouse, and rat reactivity. Its WB image uses K562 and RAW 264. cell lysates and rat kidney lysate; these examples do not establish performance in every sample type.
Which to pick: M02269 is the only listed option. Its WB image includes human K562 cells, mouse RAW 264. cells, and rat kidney; choose it if its listed reactivity and tested samples fit your experiment.
Source: BosterBio PIK3CD gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00329.
- Human Protein Atlas. PIK3CD tissue expression.
- PMC10964935 — target-verified WB comparison
- PMC13374213 — target-verified WB comparison
- PMC10589178 — target-verified WB comparison