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- Table of Contents
CAMK2G / Calcium/calmodulin-dependent protein kinase type II subunit gamma · Western blot design guide
Design a Western Blot for CAMK2G
Source-linked CAMK2G Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CAMK2G 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
CAMK2G 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 | ~62.6 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebral cortex (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 | CAMK2B cross-reactivity | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 11 isoform(s) |
Section 1
Source-Linked CAMK2G Western Blot Protocol Options
The A04046-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | U-87MG (40ug), PC3 (40ug), A549 (40ug), H9C2 (40ug), CT26 (40ug) (catalog A04046-1) |
| 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 | A04046-1; 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 CAMK2G Western Blot Band Size?
CAMK2G is predicted at 62.6 kDa; isoforms and phosphorylation could affect migration, but no empirical band size or visible shift is established.
| Band near 62.6 kDa | Compatible with the predicted CAMK2G mass; identity requires controls |
| Several bands at different positions | Could reflect splice isoforms, though distinct migration is unproven |
| Closely spaced doublet | Could reflect phosphorylation or isoforms; its cause requires testing |
| Single sharp band near 62.6 kDa | Compatible with CAMK2G even if its isoforms are unresolved |
💡Expected CAMK2G appearanceCAMK2G has a predicted mass of 62.6 kDa, but no empirical band size is supplied; confirm any candidate band with antibody specificity or CAMK2G depletion controls.
| UniProt predicted mass | Places the reference sequence near 62.6 kDa |
| Splice isoform 1 | Its individual mass and migration are not supplied |
| Splice isoforms 2 through 6 | May differ in size, but individual masses and migration are not supplied |
| Splice isoforms 7 through 11 | May differ in size, but individual masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | A splice isoform or altered migration is possible; identity is unconfirmed | Compare with CAMK2G depletion and an independent antibody |
| Band lower than expected | A splice isoform is possible; identity is unconfirmed | Check CAMK2G depletion and the antibody epitope |
| Multiple bands | Eleven annotated isoforms or phosphorylation may contribute; resolution is unproven | Use CAMK2G depletion and phosphatase treatment to identify the bands |
| Broad smear instead of sharp band | Variable phosphorylation is possible, but its effect on migration is unproven | Compare treated and untreated samples with phosphatase controls |
| Weak or no signal | Recovery of this peripheral sarcoplasmic reticulum membrane protein may be limited | Check extraction and sample loading, then validate with a positive control |
Experiment evidence
Western blot images showing CAMK2G band
A04046-1 · CAMK2G
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CAMK2G Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CAMK2G in Western blot, you can use cerebral cortex lysate, which HPA rates as highly positive.
Positive control: Cerebral cortex (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Tissue controls are feasible, but membrane-associated CAMK2G may require effective protein extraction.
HPA tissue expression evidence for CAMK2G
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 |
|---|---|---|---|---|
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CAMK2G Western Blot Tips
Deeper troubleshooting and optimisation questions for CAMK2G, answered from its protein features.
How should CAMK2G 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 CAMK2G isoforms affect band size?
Isoforms · UniProt lists 11 isoforms. For example, isoforms 3, 7, and 10 lack residues 331–364; isoforms 2, 8, 9, and 11 lack residues 331–341. Compare any candidate band with the sequence of the isoform your antibody recognizes.
Which CAMK2G phosphorylation sites are listed?
PTM · UniProt lists autocatalytic phosphothreonine at positions 287, 306, and 307, plus phosphoserine at 311, 334, 349, 352, 419, and 484. These are the supplied UniProt coordinates; antibody or paper numbering may differ. The repeated wording at position 484 does not establish additional distinct sites.
CAMK2G is a calcium/calmodulin-dependent kinase, and UniProt lists autocatalytic phosphorylation at Thr287, Thr306, and Thr307. Compare stimulated and control samples with both a site-specific phospho antibody and total CAMK2G detection. The supplied features do not establish a stimulation protocol or guarantee a signal change.
Measure the phospho-signal alongside total CAMK2G in the same conditions, then compare their ratio. Check which isoforms the antibodies recognize: several alternative sequences remove regions containing listed phosphorylation positions. A change in phospho-signal alone cannot distinguish altered phosphorylation from altered CAMK2G abundance.
CAMK2G is a calcium/calmodulin-dependent kinase, and UniProt lists autocatalytic phosphorylation at Thr287, Thr306, and Thr307. Compare stimulated and control samples with both a site-specific phospho antibody and total CAMK2G detection. The supplied features do not establish a stimulation protocol or guarantee a signal change.
Measure the phospho-signal alongside total CAMK2G in the same conditions, then compare their ratio. Check which isoforms the antibodies recognize: several alternative sequences remove regions containing listed phosphorylation positions. A change in phospho-signal alone cannot distinguish altered phosphorylation from altered CAMK2G abundance.
Does this guide establish induction of CAMK2G?
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 CAMK2G Western blot?
Transfer · Choose and verify a transfer method suitable for a protein predicted at 62.6 kDa; the supplied features do not specify transfer settings. CAMK2G is annotated as a peripheral protein on the cytoplasmic side of the sarcoplasmic reticulum membrane, so account for its location when preparing samples.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04046-1 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 CAMK2G 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.
Why might CAMK2G migrate differently from its predicted mass?
Interpretation · The canonical sequence predicts 62.6 kDa. Alternative splicing and phosphorylation are listed, but their presence alone does not establish a visible shift or explain a particular band. No observed band size is supplied.
Check antibody recognition against the 11 listed isoforms and their alternative sequences. Phosphorylation is also listed, but a feature annotation alone cannot identify a shifted band. Compare candidate bands across samples and antibody specificities before assigning an isoform or modified form.
Check antibody recognition against the 11 listed isoforms and their alternative sequences. Phosphorylation is also listed, but a feature annotation alone cannot identify a shifted band. Compare candidate bands across samples and antibody specificities before assigning an isoform or modified form.
Boster reagents
CAMK2G 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-CaMKII Beta / Gamma (R631) CAMK2G Antibody
Cat # A04046-1
A04046-1 is listed for human, mouse, and rat reactivity. Its WB image shows CaMKIIβ/γ staining at 1:500 in U-87MG, PC3, A549, H9C2, and CT26 whole-cell lysates. The supplied evidence does not establish CAMK2G-specific detection.
Which to pick: A04046-1 is the only listed antibody and has a WB validation image. Use the reported 1:500 dilution and lysate examples to judge relevance to your experiment; the CaMKIIβ/γ name does not establish gamma-only specificity.
Source: BosterBio CAMK2G gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13555.
- Human Protein Atlas. CAMK2G tissue expression.
- PMC9853378 — target-verified WB comparison
- PMC8698062 — target-verified WB comparison
- PMC6240363 — target-verified WB comparison