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SIK2 / Serine/threonine-protein kinase SIK2 · Western blot design guide
Design a Western Blot for SIK2
Source-linked SIK2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SIK2 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
SIK2 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 | ~103.9 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Bone marrow (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SIK2 Western Blot Protocol Options
The A04515 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | SW480 cell lysate (catalog A04515) |
| 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 | A04515 · 1 μg/mL (catalog A04515) |
| 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 SIK2 Western Blot Band Size?
SIK2 is predicted at 103.9 kDa; annotated modifications may affect mobility, but no empirical band size or demonstrated migration shift is supplied.
| Band near 104 kDa | Consistent with the 103.9 kDa predicted SIK2 mass; confirm identity with controls |
| Close bands near 104 kDa | Could reflect different modification states, but distinct migration is not established |
| Signal in a cytoplasmic fraction | Consistent with the annotated cytoplasmic location |
| Signal in an endoplasmic reticulum membrane fraction | Consistent with the annotated membrane location |
💡Expected SIK2 appearanceSIK2 has a predicted mass of 103.9 kDa; no empirical band size is supplied, and band identity should be checked with controls.
| Predicted SIK2 mass | Sets a 103.9 kDa reference for the full-length protein |
| Phosphothreonine at residues 25, 175, and 484 | May affect mobility; no band shift is established |
| Phosphoserine at residues 534 and 587 | May affect mobility; no band shift is established |
| N6-acetyllysine at residue 53 | Is annotated, but no band shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SIK2 may be missed if extraction poorly recovers its endoplasmic reticulum membrane pool | Check membrane protein recovery and include a SIK2-positive lysate |
| Band higher than expected | The supplied features do not establish a higher migrating SIK2 species | Check antibody specificity with SIK2 depletion and compare independent antibodies |
| Band lower than expected | The supplied features do not establish a smaller SIK2 product | Check identity with SIK2 depletion and an antibody to a different region |
| Multiple bands | Modification states are possible, but distinct SIK2 bands are not established | Use SIK2 depletion to identify which bands depend on SIK2 |
| Weak or no signal | The sampled fraction may contain little cytoplasmic or endoplasmic reticulum membrane SIK2 | Compare fraction recovery and test a SIK2-positive lysate |
Experiment evidence
Western blot images showing SIK2 band
A04515 · SIK2
A04515-1 · SIK2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SIK2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SIK2 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Bone marrow (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: SIK2 is intracellular, so tissue lysates are suitable, though the HPA positive signal is only medium.
HPA tissue expression evidence for SIK2
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | myoepithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | follicle cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SIK2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SIK2, answered from its protein features.
Where should the SIK2 band appear?
Band shift · The predicted mass is 103.9 kDa. No observed band position is supplied, so use this as a reference rather than an exact migration target.
Could multiple bands represent SIK2 isoforms?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning additional bands to annotated SIK2 isoforms.
Which SIK2 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphothreonine at positions 25, 175, and 484, and phosphoserine at 534 and 587; LKB1 is specified for position 175. These are UniProt coordinates, which may differ from antibody or paper numbering. Their presence alone does not establish a visible shift.
UniProt lists N6-acetyllysine at position 53 by EP300. Check the antibody’s epitope if signal changes with acetylation state. This site alone does not establish a detectable mass shift.
UniProt lists N6-acetyllysine at position 53 by EP300. Check the antibody’s epitope if signal changes with acetylation state. This site alone does not establish a detectable mass shift.
Does this guide establish induction of SIK2?
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 SIK2?
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 A04515 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 SIK2 signal be quantified across samples?
Quantitation · Use comparable sample fractions across conditions, since SIK2 is annotated in both the cytoplasm and at the endoplasmic reticulum membrane. If measuring phosphorylation, distinguish site-specific signal from total SIK2 signal; the listed sites do not establish how much of the protein is modified.
Which cellular fractions could contain SIK2?
Interpretation · UniProt places SIK2 in the cytoplasm and at the endoplasmic reticulum membrane. Consider both locations when choosing a fraction; a weak signal in one fraction does not by itself establish low total SIK2 abundance.
Compare them with the 103.9 kDa predicted mass and validate their identity before assigning them to SIK2. The record lists one isoform, five phosphorylation sites, and one acetylation site, but provides no observed band position or evidence that these modifications cause a visible shift.
Compare them with the 103.9 kDa predicted mass and validate their identity before assigning them to SIK2. The record lists one isoform, five phosphorylation sites, and one acetylation site, but provides no observed band position or evidence that these modifications cause a visible shift.
Boster reagents
SIK2 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-SIK2 Antibody
Cat # A04515
Real WB data
Anti-SIK2 Antibody Picoband®
Cat # A04515-1
A04515 and A04515-1 both list human, mouse, and rat reactivity and have Western blot images. A04515 is shown with SW480 lysate; A04515-1 is shown with human, rat, and mouse lysates. A04515-1 reports a band near 120 kDa versus 104 kDa expected.
Which to pick: Choose by the documented sample context: A04515 shows SW480 lysate at 1 μg/mL; A04515-1 shows U2OS, Caco-2, rat brain, PC-12, mouse brain, and 3T3-L1 lysates at 0.5 μg/mL. Both have WB images; consider A04515-1’s reported band-size difference.
Source: BosterBio SIK2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9H0K1.
- Human Protein Atlas. SIK2 tissue expression.
- PMC4973365 — target-verified WB comparison
- PMC7782074 — target-verified WB comparison
- PMC10701601 — target-verified WB comparison