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NDRG2 / Protein NDRG2 · Western blot design guide
Design a Western Blot for NDRG2
Source-linked NDRG2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NDRG2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
NDRG2 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 | ~40.8 kDa | |
| Observed band | 41 kDa | |
| Gel | 10% (catalog PB9672) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Lymph node (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 6 isoform(s) |
Section 1
Source-Linked NDRG2 Western Blot Protocol Options
The PB9672 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, rat brain, mouse brain (catalog PB9672) |
| Gel % | 10% (catalog PB9672) |
| Load | 30 ug; reducing conditions (catalog PB9672) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog PB9672) |
| Membrane | nitrocellulose membrane (catalog PB9672) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog PB9672) |
| Primary antibody | PB9672 · 0.5 μg/mL (catalog PB9672) |
| Primary incubation | overnight at 4°C (catalog PB9672) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog PB9672) |
| Secondary incubation | 1.5 hour at RT (catalog PB9672) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PB9672) |
| Detection | ECL (catalog PB9672) |
Section 2
What Is the Expected NDRG2 Western Blot Band Size?
NDRG2 is predicted at 40.8 kDa and observed at approximately 41 kDa; the small difference has no established cause in the supplied evidence.
| Band at approximately 41 kDa | Matches the observed NDRG2 band in reducing whole-cell and brain lysate blots |
| Band near 40.8 kDa | Consistent with the predicted mass; confirm identity with a control |
| Several bands near the expected region | Could include NDRG2 isoforms 1–6, whose individual masses are not supplied |
| Closely spaced doublet | Could reflect different phosphorylation states, but a migration difference is not established |
💡Expected NDRG2 appearanceNDRG2 has a predicted mass of 40.8 kDa and an observed band at approximately 41 kDa; confirm band identity with a positive control or NDRG2 depletion.
| Predicted NDRG2 mass | 40.8 kDa; the observed band is approximately 41 kDa |
| Isoform 1 | Its individual mass and migration relative to other isoforms are not supplied |
| Isoforms 2 and 3 | Their individual masses and migration relative to other isoforms are not supplied |
| Isoforms 4, 5 and 6 | Their individual masses and migration relative to other isoforms are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | Band identity or an effect of NDRG2 phosphorylation is unconfirmed | Compare with the 41 kDa control band and test NDRG2 depletion |
| Band lower than expected | An alternative isoform is possible, but its mass is not supplied | Check antibody isoform recognition and test NDRG2 depletion |
| Multiple bands | Isoforms or phosphorylation states are possible; distinct migration is unconfirmed | Use NDRG2 depletion and, if relevant, compare phosphatase-treated lysate |
| Weak or no signal | Insufficient detectable NDRG2 in the sample is possible | Check loading and compare with HeLa or brain lysate as a positive control |
| Fragments below expected size | Band identity or sample degradation is unconfirmed | Compare fresh lysate with a positive control and test NDRG2 depletion |
Experiment evidence
Western blot images showing NDRG2 band
PB9672 · NDRG2
PA2148 · NDRG2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NDRG2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NDRG2 in Western blot, you can use adrenal gland lysate, which HPA scores High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Lymph node (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Tissue lysates are suitable for cytoplasmic NDRG2, though HPA's Not detected score for lymph node does not guarantee a blank blot.
HPA tissue expression evidence for NDRG2
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 | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced NDRG2 Western Blot Tips
Deeper troubleshooting and optimisation questions for NDRG2, answered from its protein features.
What should be checked when unexpected NDRG2 bands appear?
Band shift · Check antibody specificity and epitope location against the six isoforms and their listed sequence changes. Phosphorylation is another feature to investigate, but its presence does not prove a visible shift. Compare the bands with the reported 41 kDa signal before assigning identities.
Which NDRG2 isoforms could produce different bands?
Isoforms · UniProt lists six isoforms. Isoforms 2, 4, 5 and 6 lack residues 26–39; isoform 4 also lacks 157–185, isoform 3 lacks 262–272, and isoform 5 lacks 272–287. Isoform 6 has an altered N terminus. Check which isoforms the antibody recognizes before assigning bands.
An epitope within residues 26–39 would be absent from isoforms 2, 4, 5 and 6. Residues 157–185 are absent from isoform 4; 262–272 from isoform 3; and 272–287 from isoform 5. Match the antibody epitope to these UniProt coordinates when interpreting missing bands.
An epitope within residues 26–39 would be absent from isoforms 2, 4, 5 and 6. Residues 157–185 are absent from isoform 4; 262–272 from isoform 3; and 272–287 from isoform 5. Match the antibody epitope to these UniProt coordinates when interpreting missing bands.
Could NDRG2 phosphorylation cause a band shift?
PTM · UniProt lists phosphothreonine 20 and multiple phosphoserine and phosphothreonine sites from 326 to 370. These features alone do not establish a visible shift. Compare treated and untreated samples if testing whether phosphorylation affects migration.
Report sites using the supplied UniProt coordinates, such as Thr20, Ser326 and Ser370. UniProt coordinates can differ from paper or antibody numbering, so state the numbering convention when comparing sources.
Report sites using the supplied UniProt coordinates, such as Thr20, Ser326 and Ser370. UniProt coordinates can differ from paper or antibody numbering, so state the numbering convention when comparing sources.
Does this guide establish induction of NDRG2?
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 NDRG2?
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 PB9672 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 multiple NDRG2 bands be quantified?
Quantitation · Define the band or bands being measured and use the same rule across samples. Six isoforms and numerous phosphorylation sites are listed, but those features alone do not identify each band. Avoid combining bands as total NDRG2 without confirming what the antibody detects.
How does the 41 kDa band compare with predicted NDRG2 mass?
Interpretation · The reported 41 kDa band is close to the 40.8 kDa predicted mass. This small difference does not require a modification-based explanation.
Boster reagents
NDRG2 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-NDRG2 Antibody Picoband®
Cat # PB9672
Real WB data
Anti-Protein NDRG2 NDRG2 Antibody Picoband®
Cat # PA2148
PB9672 and PA2148 are listed for human, mouse, and rat NDRG2. Each has a Western blot image showing an approximately 41 kDa band in the samples tested. The supplied evidence is limited to these product blots.
Which to pick: Choose PB9672 for a documented HeLa, rat brain, or mouse brain blot; choose PA2148 for HepG2 or rat and mouse brain or heart blots. Both images used 30 μg of lysate and 0.5 μg/mL primary antibody.
Source: BosterBio NDRG2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UN36.
- Human Protein Atlas. NDRG2 tissue expression.
- PMC2922092 — target-verified WB comparison
- PMC5042018 — target-verified WB comparison
- PMC11556958 — target-verified WB comparison