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NLRP12 / NACHT, LRR and PYD domains-containing protein 12 · Western blot design guide
Design a Western Blot for NLRP12
Real validated NLRP12 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NLRP12 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
NLRP12 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 | ~120.2 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Fallopian tube (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 | 7 isoform(s) |
Section 1
Real Curated NLRP12 Western Blot Protocols
The A02124 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human brain tissue lysate (catalog A02124) |
| 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 | A02124 · 1 μg/mL (catalog A02124) |
| 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 NLRP12 Western Blot Band Size?
NLRP12 is predicted at 120.2 kDa; seven isoforms could affect band patterns, but no empirical migration or resolvable isoform sizes are supplied.
| Band near 120.2 kDa | Consistent with the predicted NLRP12 mass; identity requires controls |
| Several bands at different positions | Could reflect expression of multiple NLRP12 splice isoforms |
| Band below 120.2 kDa | Could represent a shorter splice isoform; its mass is unknown |
| Band above 120.2 kDa | Could represent a larger splice isoform; its mass is unknown |
💡Expected NLRP12 appearanceUniProt predicts NLRP12 at 120.2 kDa; no empirical band size or isoform migration is supplied, so confirm any candidate band with antibody specificity controls.
| Predicted NLRP12 mass | Provides a 120.2 kDa reference, not a measured band position |
| Splice isoform 1 | Its individual migration is not supplied |
| Splice isoforms 2, 3, and 4 | May differ in size from other isoforms; relative masses are not supplied |
| Splice isoforms 5, 6, and 7 | May differ in size from other isoforms; relative masses are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | NLRP12 may be insufficiently abundant in the sampled lysate | Check loading and use a confirmed NLRP12-positive lysate |
| Band higher than expected | A splice isoform or an unrelated protein could produce the band | Confirm identity by NLRP12 knockdown or a second antibody |
| Band lower than expected | A shorter splice isoform or an unrelated protein could produce the band | Check isoform expression and confirm identity by NLRP12 knockdown |
| Multiple bands | Seven annotated isoforms could contribute, but their migration is unknown | Compare isoform expression and validate candidate bands with NLRP12 knockdown |
| Weak or no signal | Low NLRP12 abundance or insufficient antibody sensitivity | Verify loading and compare with a confirmed NLRP12-positive control |
Experiment evidence
Western blot images showing NLRP12 band
A02124 · NLRP12
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NLRP12 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NLRP12 in Western blot, you can use fallopian tube tissue, which has high HPA expression.
Positive control: Fallopian tube (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: As a cytoplasmic protein, NLRP12 can be assessed in tissue lysate; adipose tissue provides an HPA not-detected comparison.
HPA tissue expression evidence for NLRP12
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 |
|---|---|---|---|---|
| Fallopian tube | glandular cells | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Urinary bladder | urothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NLRP12 Western Blot Tips
Deeper troubleshooting and optimisation questions for NLRP12, answered from its protein features.
What band size should I expect for NLRP12?
Band shift · The canonical sequence is 1,061 residues with a predicted mass of 120.2 kDa. No observed band position is supplied. Use 120.2 kDa as a reference, not a guaranteed apparent mass; the listed sequence variants alone do not establish a visible shift.
Could NLRP12 isoforms produce different bands?
Isoforms · Seven isoforms are listed. Relative to the canonical sequence, isoform 5 lacks residues 1–717 and 921–977 and replaces residues 718–748. Isoforms 2, 3, 4, and 6 have shorter listed deletions. These changes could affect size, but the features do not establish which isoforms produce detectable bands.
Check the antibody epitope against the listed sequence changes. For example, an epitope within canonical residues 862–1031 is absent from isoform 4, while one within residues 1–717 is absent from isoform 5. Coordinates here refer to the supplied UniProt canonical sequence; antibody numbering may differ.
Check the antibody epitope against the listed sequence changes. For example, an epitope within canonical residues 862–1031 is absent from isoform 4, while one within residues 1–717 is absent from isoform 5. Coordinates here refer to the supplied UniProt canonical sequence; antibody numbering may differ.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of NLRP12?
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 NLRP12 Western blot?
Transfer · Choose and verify transfer conditions suitable for a protein near the canonical predicted mass of 120.2 kDa. Check transfer with a marker around that size and inspect the gel for retained protein. The supplied features do not specify a membrane or transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02124 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 I quantify NLRP12 bands across samples?
Quantitation · Quantify the same defined band or band set across samples, and document which bands the antibody detects. Seven listed isoforms include substantial sequence differences, so a single band may not represent every isoform. The supplied features give no observed band position to assign bands by size alone.
How should I interpret unexpected NLRP12 bands?
Interpretation · Compare their sizes with the 120.2 kDa canonical prediction, then check whether the antibody epitope is retained in the listed isoforms. Isoform 7 has an L-to-LR change at canonical position 691; several others contain deletions. Those features provide candidates to investigate, but do not identify an unexpected band on their own.
Boster reagents
NLRP12 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-NALP12 NLRP12 Antibody
Cat # A02124
the supplier A02124 is an anti-NLRP12 antibody listed for human, mouse, and rat. Its WB image shows NLRP12 analysis in human brain tissue lysate at 1 μg/mL; the supplied caption does not show mouse or rat WB samples.
Which to pick: A02124 is the only listed option. It has a WB image using human brain tissue lysate. For mouse or rat samples, the catalog lists reactivity, but the supplied WB evidence is from human tissue.
Source: BosterBio NLRP12 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P59046.
- Human Protein Atlas. NLRP12 tissue expression.
- PMC11306119 — target-verified WB comparison
- PMC6483596 — target-verified WB comparison
- PMC10611651 — target-verified WB comparison