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- Table of Contents
PRKCQ / Protein kinase C theta type · Western blot design guide
Design a Western Blot for PRKCQ
Source-linked PRKCQ Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PRKCQ 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
PRKCQ 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 | ~81.9 kDa | |
| Observed band | ~82 kDa | |
| Gel | 8–10% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked PRKCQ Western Blot Protocol Options
The M01293 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog M01293) |
| Gel % | 8–10% (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 | M01293; 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 PRKCQ Western Blot Band Size?
PRKCQ is predicted at 81.9 kDa and observed at ~82 kDa; the small difference has no established cause in the supplied evidence.
| Band at ~82 kDa | Matches the empirical PRKCQ band and its 81.9 kDa predicted mass; confirm identity with controls |
| Additional bands near the main band | Isoforms 1, 2 and 3 or phosphorylation may contribute, but distinct migration is unproven |
| Weak band in a cytosolic fraction after TCR stimulation | PRKCQ may have moved to membrane lipid rafts |
| Band in a membrane fraction after TCR stimulation | Consistent with PRKCQ recruitment to membrane lipid rafts |
💡Expected PRKCQ appearancePRKCQ has a predicted mass of 81.9 kDa and an empirical band at ~82 kDa; confirm band identity with a positive lysate and a PRKCQ-specific control.
| Predicted PRKCQ mass | 81.9 kDa predicted; the empirical band is ~82 kDa |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its size relative to isoform 1 and its migration are not supplied |
| Isoform 3 | Its size relative to the other isoforms and its migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PRKCQ abundance or detection may be insufficient | Run Jurkat lysate as a positive control and check loading and transfer |
| Band higher than expected | An isoform or phosphorylation could affect migration, but the cause is unestablished | Compare with the ~82 kDa Jurkat band and verify identity by PRKCQ depletion |
| Band lower than expected | An isoform or an unrelated band could be present; no fragment size is established | Verify identity by PRKCQ depletion and compare with a positive lysate |
| Multiple bands | PRKCQ has three isoforms and documented phosphorylation sites, but distinct bands are unproven | Check which bands disappear after PRKCQ depletion |
| Weak or no signal | PRKCQ can redistribute from cytoplasm to membrane lipid rafts after TCR stimulation | Compare total lysate and fractions, with Jurkat lysate as a positive control |
| Fragments below expected size | Degradation or nonspecific binding is possible; no cleavage feature is supplied | Use fresh lysate with protease inhibitors and verify bands by PRKCQ depletion |
Experiment evidence
Western blot images showing PRKCQ band
M01293 · PRKCQ
M01293 · PRKCQ
A01293-3 · PRKCQ
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PRKCQ Western blot
🧪For positive controls for PRKCQ in Western blot, you can use T-cell lysate, although the supplied HPA data identify no top positive sample.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no tissue data here, so a validated positive tissue and clean negative tissue cannot be selected.
HPA tissue expression evidence for PRKCQ
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced PRKCQ Western Blot Tips
Deeper troubleshooting and optimisation questions for PRKCQ, answered from its protein features.
How should PRKCQ 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 isoform 2 produce a smaller band?
Isoforms · Isoform 2 lacks canonical residues 550–612. A band below the canonical ~82 kDa band could be consistent with this shorter sequence, but band position alone cannot identify the isoform.
Isoform 3 lacks canonical residues 2–126. When assessing a lower band, consider whether the antibody recognizes a region retained in isoform 3; the supplied features do not specify an antibody epitope.
Isoform 3 lacks canonical residues 2–126. When assessing a lower band, consider whether the antibody recognizes a region retained in isoform 3; the supplied features do not specify an antibody epitope.
Which PRKCQ phosphorylation sites matter when interpreting bands?
PTM · UniProt annotates Tyr90 phosphorylated by LCK, Thr219 by autocatalysis, Ser348, Thr538 by PDPK1, and Ser676, Ser685 and Ser695. These are UniProt coordinates; antibody or paper numbering may differ. Their annotation does not establish a visible band shift.
What changes after TCR stimulation?
Induction · In resting T cells, PRKCQ is mostly cytoplasmic. After TCR stimulation, it associates with lipid rafts and localizes to the immunological synapse. Compare matched fractions if measuring its distribution; this localization change alone does not establish increased total PRKCQ.
How should transfer be checked for PRKCQ?
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 M01293 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 PRKCQ be quantified across cell fractions?
Quantitation · PRKCQ is annotated in the cytoplasm and as a peripheral cell membrane protein. Because TCR stimulation changes its localization, quantify like fractions across samples when assessing redistribution, and use whole cell samples when assessing total abundance.
Should PRKCQ run above its predicted mass?
Interpretation · The predicted mass is 81.9 kDa, close to the observed ~82 kDa band. Phosphorylation is annotated, but its presence alone does not establish a visible shift or explain any difference in apparent mass.
Check its position relative to the ~82 kDa observed band and whether the antibody's target region is present in isoforms 2 and 3. Isoform 2 lacks residues 550–612; isoform 3 lacks residues 2–126. The annotated phosphorylation sites alone cannot identify an unexpected band.
Check its position relative to the ~82 kDa observed band and whether the antibody's target region is present in isoforms 2 and 3. Isoform 2 lacks residues 550–612; isoform 3 lacks residues 2–126. The annotated phosphorylation sites alone cannot identify an unexpected band.
Boster reagents
PRKCQ 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-PKC theta Rabbit Monoclonal Antibody
Cat # M01293
Real WB data
Anti-PKC theta/PRKCQ Antibody Picoband®
Cat # A01293-3
Two the supplier anti-PRKCQ antibodies have Western blot images: M01293 in Jurkat cell lysate and A01293-3 in human K562 whole-cell and blood lysates, with an approximately 78 kDa band reported for A01293-3. No publication or independent validation evidence is supplied.
Which to pick: For human K562 or blood lysates, A01293-3 has matching WB examples and reported conditions. For Jurkat lysate, M01293 has a WB example and lists human, mouse, and rat reactivity; its supplied image documents Jurkat only.
Source: BosterBio PRKCQ gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q04759.
- Human Protein Atlas. PRKCQ tissue expression.
- PMC12841367 — target-verified WB comparison
- PMC11610359 — target-verified WB comparison
- PMC5085156 — target-verified WB comparison