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- Table of Contents
CMPK2 / UMP-CMP kinase 2, mitochondrial · Western blot design guide
Design a Western Blot for CMPK2
Source-linked CMPK2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CMPK2 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
CMPK2 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 | ~49.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked CMPK2 Western Blot Protocol Options
The A11251 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat lung tissue lysate (catalog A11251) |
| 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 | A11251 · 1 μg/mL (catalog A11251) |
| 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 CMPK2 Western Blot Band Size?
CMPK2 has a predicted mass of 49.4 kDa; four isoforms could affect migration, but no specific band position or isoform pattern is demonstrated.
| Single band near 49.4 kDa | Consistent with the predicted CMPK2 mass; confirm identity with controls |
| Several discrete bands | Could reflect isoforms 1–4 if their migration differs; band identities require confirmation |
| Band away from 49.4 kDa | An isoform is possible, but its mass and migration are not supplied |
| Weak band in whole-cell lysate | Mitochondrial CMPK2 may be easier to detect in a mitochondrial fraction |
💡Expected CMPK2 appearanceUniProt predicts 49.4 kDa for CMPK2; no empirical band size or isoform masses are supplied, so confirm any candidate band with ordinary identity controls.
| UniProt predicted mass | Sets a 49.4 kDa reference, not a validated migration position |
| Splice isoform 1 | Apparent size relative to other isoforms is unknown |
| Splice isoform 2 | Could migrate differently; its mass is not supplied |
| Splice isoform 3 | Could migrate differently; its mass is not supplied |
| Splice isoform 4 | Could migrate differently; its mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Mitochondrial CMPK2 may be depleted during sample preparation | Check a mitochondrial fraction and sample recovery |
| Band higher than expected | An isoform or unrelated antibody target is possible | Compare with a CMPK2 depletion or knockout control |
| Band lower than expected | An isoform or unrelated antibody target is possible | Confirm identity with CMPK2 depletion or knockout |
| Multiple bands | Four annotated isoforms could contribute, but their migration is unknown | Compare bands with CMPK2 depletion or knockout |
| Weak or no signal | Mitochondrial CMPK2 may be poorly represented in the sample | Check loading and compare with a mitochondrial fraction |
Experiment evidence
Western blot images showing CMPK2 band
A11251 · CMPK2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CMPK2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CMPK2 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (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: CMPK2 is mitochondrial, so mitochondrial enrichment may improve detection.
HPA tissue expression evidence for CMPK2
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 | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal 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 → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
Section 3
Advanced CMPK2 Western Blot Tips
Deeper troubleshooting and optimisation questions for CMPK2, answered from its protein features.
How should CMPK2 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 CMPK2 isoforms produce different bands?
Isoforms · Four isoforms are listed, with sequence changes that could affect size. Isoform 2 replaces residues 280–303 and lacks residues 304–449; isoform 3 replaces residues 332–449; isoform 4 lacks residues 410–449. These features do not establish which isoforms are present or where their bands migrate.
Does CMPK2 have a documented modification that explains an extra band?
PTM · No modified residues or glycosylation sites are listed in the supplied features. An extra band cannot be assigned to a specific modification from this evidence. Consider the documented isoforms, but do not identify a band by size alone.
Does this guide establish induction of CMPK2?
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 CMPK2?
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 A11251 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.
What matters when quantifying CMPK2 across samples?
Quantitation · Use a consistent band and sample fraction across comparisons. Four isoforms are listed, and CMPK2 is mitochondrial; a change in which isoform or fraction is measured could change what the signal represents. The supplied features give no observed band position for selecting a quantification band.
Should CMPK2 migrate at its predicted 49.4 kDa?
Interpretation · 49.4 kDa is the predicted mass of the 449-residue canonical sequence. No observed band position is supplied. The transit peptide annotation and alternative isoforms do not establish a visible shift or explain a difference from the predicted mass.
Consider the documented alternative sequences when assessing band size, especially the missing regions in isoforms 2 and 4. A smaller band alone cannot identify an isoform; compare it with an isoform-specific sequence or detection strategy.
CMPK2 has a transit peptide annotation and is localized to mitochondria. No cleavage coordinate or processed mass is supplied, so the annotation alone cannot predict a processed band or its migration.
CMPK2 is documented in mitochondria, and mitochondrial localization is required for its antiviral function. When comparing fractions, track which fraction each band came from. Localization supports the expected compartment but does not identify an unexpected band by itself.
Consider the documented alternative sequences when assessing band size, especially the missing regions in isoforms 2 and 4. A smaller band alone cannot identify an isoform; compare it with an isoform-specific sequence or detection strategy.
CMPK2 has a transit peptide annotation and is localized to mitochondria. No cleavage coordinate or processed mass is supplied, so the annotation alone cannot predict a processed band or its migration.
CMPK2 is documented in mitochondria, and mitochondrial localization is required for its antiviral function. When comparing fractions, track which fraction each band came from. Localization supports the expected compartment but does not identify an unexpected band by itself.
Boster reagents
CMPK2 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-CMPK2 Antibody
Cat # A11251
The catalog reports one anti-CMPK2 antibody for Western blotting. A11251 is listed as reactive with human, mouse, and rat; its WB image shows rat lung tissue lysate tested at 1 μg/mL. No publication evidence is supplied.
Which to pick: A11251 is the only listed option. Its WB image documents use in rat lung tissue lysate; the supplied evidence does not show WB results for human or mouse samples.
Source: BosterBio CMPK2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q5EBM0.
- Human Protein Atlas. CMPK2 tissue expression.
- PMC11300820 — target-verified WB comparison
- PMC8515557 — target-verified WB comparison
- PMC10058120 — target-verified WB comparison