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SGK3 / Serine/threonine-protein kinase Sgk3 · Western blot design guide
Design a Western Blot for SGK3
Source-linked SGK3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SGK3 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
SGK3 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 | ~57.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked SGK3 Western Blot Protocol Options
The A02534 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A02534) |
| 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 | A02534; 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 SGK3 Western Blot Band Size?
SGK3 has a predicted mass of 57.1 kDa; isoforms may affect migration, but no empirical band size or distinct isoform bands are established.
| Band near 57.1 kDa | Consistent with the predicted SGK3 mass; confirm identity with controls |
| Single band near 57.1 kDa | Isoforms 1 and 2 need not resolve into separate bands |
| Two bands at different positions | Could represent isoforms 1 and 2; their migration is not established |
| Band away from 57.1 kDa | Could be an isoform of unknown mass; verify its identity |
💡Expected SGK3 appearanceUniProt predicts 57.1 kDa for SGK3; no empirical band size is supplied, so confirm any detected band with ordinary identity controls.
| UniProt predicted mass | Sets a 57.1 kDa reference for SGK3 |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing of isoforms 1 and 2 | Could alter band position, but a resolvable difference is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SGK3 may be scarce or poorly recovered from endosomal material | Check lysate recovery and run an SGK3 positive control |
| Band higher than expected | An isoform or unrelated antibody target is possible; the cause is unverified | Compare a positive control and confirm identity by SGK3 depletion |
| Band lower than expected | An isoform or unrelated antibody target is possible; no isoform mass is supplied | Confirm identity by SGK3 depletion and compare a positive control |
| Multiple bands | Isoforms 1 and 2 may contribute, but distinct migration is unproven | Use SGK3 depletion to identify specific bands |
| Weak or no signal | SGK3 abundance or recovery may be low in the tested sample | Check loading and antibody performance with a positive control |
Experiment evidence
Western blot images showing SGK3 band
A02534 · SGK3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SGK3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SGK3 in Western blot, you can use appendix tissue, which HPA scores High.
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: HPA identifies adipose tissue as a feasible negative control, and SGK3 is localized to endosomes.
HPA tissue expression evidence for SGK3
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 | endocrine cells | High | Protein (IHC) | HPA → |
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Lung | macrophages | 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 → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SGK3 Western Blot Tips
Deeper troubleshooting and optimisation questions for SGK3, answered from its protein features.
How should SGK3 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 SGK3 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 327–358 of the canonical sequence, so it may migrate differently. A second band alone cannot establish isoform identity.
Which SGK3 phosphorylation sites should I track?
PTM · The supplied UniProt features list Ser-126, Ser-129, Thr-320, and Ser-486 as phosphorylated residues; Thr-320 is phosphorylated by PDPK1. These are UniProt canonical-sequence coordinates, which may differ from antibody or paper numbering.
SGK3 localizes to early and recycling endosomes. Its UniProt note says endosomal localization is required for complete kinase activity and for colocalization with the kinase responsible for Ser-486 phosphorylation, enabling PDPK1 phosphorylation of Thr-320. Consider localization when interpreting changes in phospho-SGK3 signal.
Measure the phosphosite signal alongside total SGK3 so changes in protein abundance can be distinguished from changes in phosphorylation. Specify whether the readout concerns Ser-126, Ser-129, Thr-320, or Ser-486, using the supplied UniProt canonical-sequence coordinates.
SGK3 localizes to early and recycling endosomes. Its UniProt note says endosomal localization is required for complete kinase activity and for colocalization with the kinase responsible for Ser-486 phosphorylation, enabling PDPK1 phosphorylation of Thr-320. Consider localization when interpreting changes in phospho-SGK3 signal.
Measure the phosphosite signal alongside total SGK3 so changes in protein abundance can be distinguished from changes in phosphorylation. Specify whether the readout concerns Ser-126, Ser-129, Thr-320, or Ser-486, using the supplied UniProt canonical-sequence coordinates.
Does this guide establish induction of SGK3?
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 SGK3?
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 A02534 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 SGK3 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Should SGK3 migrate at its predicted 57.1 kDa?
Interpretation · 57.1 kDa is the predicted mass of canonical SGK3. No observed band position is supplied. The listed phosphorylation sites and isoform difference alone do not establish a visible shift or explain any difference from 57.1 kDa.
Compare it with the predicted 57.1 kDa canonical protein and consider isoform 2, which lacks canonical residues 327–358. The listed phosphorylation sites are also relevant, but their presence alone does not establish a visible shift. With no observed band position supplied, these features cannot identify an unexpected band.
Compare it with the predicted 57.1 kDa canonical protein and consider isoform 2, which lacks canonical residues 327–358. The listed phosphorylation sites are also relevant, but their presence alone does not establish a visible shift. With no observed band position supplied, these features cannot identify an unexpected band.
Boster reagents
SGK3 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-SGK3 Antibody
Cat # A02534
A02534 is an anti-SGK3 antibody listed for Western blotting with reported Human and Mouse reactivity. Its WB image shows analysis of extracts from various cell lines; the supplied description does not identify the lines or provide experimental conditions.
Which to pick: A02534 is the only listed SGK3 antibody. It has a WB image and reported Human and Mouse reactivity; check whether its documented sample and conditions fit your experiment.
Source: BosterBio SGK3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96BR1.
- Human Protein Atlas. SGK3 tissue expression.
- PMC10285316 — target-verified WB comparison
- PMC8751936 — target-verified WB comparison
- PMC10758069 — target-verified WB comparison