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- Table of Contents
SARM1 / NAD(+) hydrolase SARM1 · Western blot design guide
Design a Western Blot for SARM1
Source-linked SARM1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SARM1 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
SARM1 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 | ~79.4 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 | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked SARM1 Western Blot Protocol Options
The A03633 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Daudi cell lysate (catalog A03633) |
| 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 | A03633 · (A) 0.5 and (B) 1 μg/mL (catalog A03633) |
| 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 SARM1 Western Blot Band Size?
SARM1 has a predicted mass of 79.4 kDa; isoforms and homooctamer formation could affect patterns, but their effects on Western-blot migration are unconfirmed.
| Band near 79.4 kDa | consistent with the predicted SARM1 protein; confirm identity with antibody controls |
| Additional band at a different size | could reflect isoform 1 or 2; their migration difference is unknown |
| High molecular weight band | could reflect retained SARM1 homooctamers if denaturation is incomplete |
| Band enriched in a cytoplasmic or mitochondrial fraction | consistent with SARM1 localization |
💡Expected SARM1 appearanceUniProt predicts 79.4 kDa for SARM1; no empirical band size is supplied, so confirm a band near that size with ordinary band-identity controls.
| Predicted molecular mass | 79.4 kDa is the reference size |
| Isoform 1 | may differ in size from isoform 2; no isoform mass is supplied |
| Isoform 2 | may differ in size from isoform 1; a distinct band is unconfirmed |
| Phosphoserine 548 | no visible migration effect is established |
| Phosphoserine 558 | no visible migration effect is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | the sampled lysate may have little SARM1 | check expression and include a positive lysate control |
| Band higher than expected | SARM1 homooctamers may persist after incomplete denaturation | compare fully denatured samples and verify band identity |
| Band lower than expected | an alternative SARM1 isoform is possible | check isoform expression and verify band identity |
| Multiple bands | SARM1 has isoforms 1 and 2, but distinct migration is unconfirmed | compare isoform expression and use antibody specificity controls |
| Weak or no signal | SARM1 abundance may be low in the sampled fraction | check cytoplasmic and mitochondrial fractions with a positive control |
Experiment evidence
Western blot images showing SARM1 band
A03633 · SARM1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SARM1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SARM1 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 such as Ponceau alongside the samples.
⚠️Feasibility: HPA rates adrenal gland expression as medium, so the positive signal may be modest.
HPA tissue expression evidence for SARM1
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 → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Medium | Protein (IHC) | HPA → |
| Kidney | cells in tubules | 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 → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SARM1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SARM1, answered from its protein features.
Where should the main SARM1 band appear?
Band shift · The canonical sequence has a predicted mass of 79.4 kDa. No observed Western blot band is supplied, so use 79.4 kDa as a reference, not a guaranteed apparent position.
Could SARM1 isoforms produce different bands?
Isoforms · Two isoforms are listed. Isoform 2 replaces the canonical residues 1–106 with a different N-terminal sequence. Consider isoform identity when interpreting bands, but the supplied features do not establish distinct apparent band positions.
Which SARM1 phosphorylation sites matter for blot interpretation?
PTM · UniProt lists phosphoserine at positions 548 and 558. These are UniProt coordinates; paper or antibody numbering may differ. Check the numbering convention before comparing site-specific results.
Phosphoserine is listed at UniProt positions 548 and 558, but site annotation alone does not show that either modification produces a visible shift or explains a difference from 79.4 kDa.
If measuring phosphorylation at UniProt Ser548 or Ser558, identify the site and numbering convention, and compare its signal with total SARM1 in the same samples. A total SARM1 band alone does not measure phosphorylation at either site.
Phosphoserine is listed at UniProt positions 548 and 558, but site annotation alone does not show that either modification produces a visible shift or explains a difference from 79.4 kDa.
If measuring phosphorylation at UniProt Ser548 or Ser558, identify the site and numbering convention, and compare its signal with total SARM1 in the same samples. A total SARM1 band alone does not measure phosphorylation at either site.
Does this guide establish induction of SARM1?
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 SARM1?
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 A03633 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 SARM1 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.
Could a high-mass band represent a SARM1 oligomer?
Interpretation · SARM1 is annotated as a homooctamer that forms a ring through its SAM domains. An unexpected high-mass band could prompt an oligomer check, but this feature does not establish that an oligomer survives the blot conditions or identify the band.
The supplied features list two isoforms with different N-terminal sequences, but no signal peptide or propeptide. Compare the band with the expected isoforms and verify its identity; these features alone cannot assign a smaller band to processing or degradation.
The supplied features list two isoforms with different N-terminal sequences, but no signal peptide or propeptide. Compare the band with the expected isoforms and verify its identity; these features alone cannot assign a smaller band to processing or degradation.
Boster reagents
SARM1 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-SARM SARM1 Antibody
Cat # A03633
The catalog reports one anti-SARM1 antibody for Western blot, A03633, with stated human and mouse reactivity. Its WB image shows Daudi cell lysate tested at 0.5 and 1 μg/mL; the supplied evidence does not show other specimens.
Which to pick: A03633 is the only listed option. Its Daudi cell lysate WB image provides a starting point for that sample context; check suitability for your intended specimen, especially mouse samples, which are listed as reactive but not shown in the supplied image.
Source: BosterBio SARM1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q6SZW1.
- Human Protein Atlas. SARM1 tissue expression.
- PMC6446871 — target-verified WB comparison
- PMC10113485 — target-verified WB comparison
- PMC4783152 — target-verified WB comparison