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- Table of Contents
NME2 / Nucleoside diphosphate kinase B · Western blot design guide
Design a Western Blot for NME2
Real validated NME2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NME2 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
NME2 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~17.3 kDa | |
| Observed band | ~17 kDa | |
| Gel | 12% (catalog M01762-1) | |
| Positive control | Endometrium (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 C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated NME2 Western Blot Protocols
The M01762-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human LNCAP, human 293T, human PC-3 (catalog M01762-1) |
| Gel % | 12% (catalog M01762-1) |
| Load | 30 ug; reducing conditions (catalog M01762-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M01762-1) |
| Membrane | nitrocellulose membrane (catalog M01762-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M01762-1) |
| Primary antibody | M01762-1 · 1:500 (catalog M01762-1) |
| Primary incubation | overnight at 4°C (catalog M01762-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M01762-1) |
| Secondary incubation | 1.5 hour at RT (catalog M01762-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M01762-1) |
| Detection | ECL (catalog M01762-1) |
Section 2
What Is the Expected NME2 Western Blot Band Size?
NME2 is predicted at 17.3 kDa and observed at ~17 kDa; the small difference has no established cause in the supplied evidence.
| Band at ~17 kDa in whole-cell lysate | Matches the empirical NME2 band near its 17.3 kDa predicted mass |
| Band near 17 kDa in a cytoplasmic fraction | Consistent with NME2's cytoplasmic location |
| Band near 17 kDa in a nuclear fraction | Consistent with NME2's nuclear location |
| Additional bands at other sizes | Could reflect isoforms 1 and 3, but their migration is not established |
💡Expected NME2 appearanceNME2 is predicted at 17.3 kDa, and antibody QC shows a band at ~17 kDa in reducing whole-cell lysates; confirm identity with an appropriate specificity control.
| UniProt predicted mass | Places the reference band near 17.3 kDa, consistent with the observed ~17 kDa band |
| 152-residue sequence | Underlies the supplied full-length predicted mass; no cleavage is annotated |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 3 | Its size may differ from isoform 1, but distinct bands are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Insufficient NME2 signal or loss during transfer | Check loading and transfer, then use a known positive lysate |
| Band higher than expected | Band identity is uncertain; no size-increasing modification is annotated | Check reduction and denaturation, then verify specificity by NME2 depletion |
| Band lower than expected | Degradation or an unrelated antibody-reactive protein | Prepare fresh lysate with protease inhibitors and verify specificity by NME2 depletion |
| Multiple bands | Isoforms 1 and 3 are annotated, but their band positions are unknown | Use isoform-specific evidence or NME2 depletion to assign bands |
| Weak or no signal | Low NME2 abundance, poor transfer or inadequate antibody signal | Check loading and transfer and compare with a known positive lysate |
| Fragments below expected size | Possible sample degradation; no physiological cleavage is annotated | Repeat with fresh, protease-inhibited lysate and verify fragment identity |
Experiment evidence
Western blot images showing NME2 band
M01762-1 · NME2
PA1830 · NME2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NME2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NME2 in Western blot, you can use endometrium tissue, which HPA rates high for NME2 expression.
Positive control: Endometrium (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: HPA lists adipose tissue as not detected, but confirm the absence of signal in your lysate.
HPA tissue expression evidence for NME2
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 |
|---|---|---|---|---|
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Prostate | glandular cells | 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 → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Breast | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced NME2 Western Blot Tips
Deeper troubleshooting and optimisation questions for NME2, answered from its protein features.
How should NME2 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 NME2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 3. Isoform 3 has an alternative sequence beginning at UniProt position 1, so its migration and antibody recognition may differ from the canonical protein. Check whether the antibody recognizes both sequences before assigning bands to isoforms.
Do the supplied features support a modification-induced band shift?
PTM · No modified residues or glycosylation sites are listed in the supplied features. Those annotations therefore provide no specific modification or site to assign to a shifted band. Compare the band's behavior with antibody specificity and sample conditions before interpreting it.
Does this guide establish induction of NME2?
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 NME2 Western blot?
Transfer · NME2 is a small protein with a predicted mass of 17.3 kDa. Choose transfer conditions that retain proteins near 17 kDa, and check the membrane and post-transfer gel if the band is weak or missing.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M01762-1 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 NME2 be quantified across cellular fractions?
Quantitation · NME2 is listed in the cytoplasm, nucleus, lamellipodium and ruffle. Quantify bands within the assay's linear range and compare like fractions, since changes in fraction composition can change the measured NME2 signal.
Why might NME2 run near 17 kDa?
Interpretation · The canonical sequence has a predicted mass of 17.3 kDa, close to the observed band near 17 kDa. The supplied features do not establish a modification or processing event that explains any small difference between calculated and apparent mass.
UniProt describes NME2 homohexamers and mixed hexamers with NME1. A higher band could prompt a check of sample preparation and antibody specificity, but the assembly annotation alone does not establish that a complex survives Western blot preparation.
UniProt describes NME2 homohexamers and mixed hexamers with NME1. A higher band could prompt a check of sample preparation and antibody specificity, but the assembly annotation alone does not establish that a complex survives Western blot preparation.
Boster reagents
NME2 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-NME2 Rabbit Monoclonal Antibody
Cat # M01762-1
Real WB data
Anti-NME2 Antibody Picoband®
Cat # PA1830
Two the supplier anti-NME2 antibodies have Western blot images showing a band near the expected 17 kDa. Both list human, mouse, and rat reactivity, but the pictured samples differ; these examples do not establish performance in every tissue or cell type.
Which to pick: Choose M01762-1 for the pictured human cell lines or rat and mouse liver and cell lysates. Choose PA1830 for the pictured rat and mouse heart or brain lysates. PA1830 lists human reactivity, but its supplied blot shows only rat and mouse samples.
Source: BosterBio NME2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P22392.
- Human Protein Atlas. NME2 tissue expression.
- PMC8502175 — target-verified WB comparison
- PMC9509583 — target-verified WB comparison
- PMC12150935 — target-verified WB comparison