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- Table of Contents
ASIC1 / Acid-sensing ion channel 1 · Western blot design guide
Design a Western Blot for ASIC1
Source-linked ASIC1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ASIC1 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
ASIC1 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 | ~59.9 kDa | |
| Observed band | ~60 kDa | |
| Gel | 5–20% (catalog A02332-2) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked ASIC1 Western Blot Protocol Options
The A02332-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human THP-1 (catalog A02332-2) |
| Gel % | 5–20% (catalog A02332-2) |
| Load | 50ug; reducing conditions (catalog A02332-2) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A02332-2) |
| Membrane | Nitrocellulose membrane (catalog A02332-2) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A02332-2) |
| Primary antibody | A02332-2 · 0.5 μg/mL (catalog A02332-2) |
| Primary incubation | overnight at 4°C (catalog A02332-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog A02332-2) |
| Secondary incubation | 1.5 hour at RT (catalog A02332-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02332-2) |
| Detection | ECL (catalog A02332-2) |
Section 2
What Is the Expected ASIC1 Western Blot Band Size?
ASIC1 is predicted at 59.9 kDa and observed near 60 kDa; glycosylation and isoforms are documented, but no specific migration effect is established.
| Band near 60 kDa | Matches the approximately 60 kDa empirical ASIC1 band and 59.9 kDa predicted mass |
| Band above 60 kDa | N-linked glycosylation at Asn368 or Asn395 could affect migration; the cause of a higher band needs confirmation |
| Multiple bands at different positions | Asic1a, 1 and Asic1b are named isoforms, but distinct band positions are not established |
| Weak band in a soluble fraction | ASIC1 is a multi-pass cell-membrane protein that may be poorly recovered in that fraction |
💡Expected ASIC1 appearanceASIC1 has a predicted mass of 59.9 kDa and an empirical band near 60 kDa; confirm band identity with an appropriate antibody or ASIC1 depletion control.
| Predicted ASIC1 mass | 59.9 kDa provides the sequence-based reference; the empirical band is near 60 kDa |
| N-linked glycosylation at Asn368 | Could affect apparent size, but its visible effect is not established |
| N-linked glycosylation at Asn395 | Could affect apparent size, but its visible effect is not established |
| Asic1a, 1 and Asic1b isoforms | May differ in size; their relative masses and band separation are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated ASIC1 may be poorly recovered during extraction | Check membrane-protein recovery and test an enriched membrane fraction |
| Band higher than expected | N-linked glycosylation could alter migration, but the band's identity and cause are unconfirmed | Compare with a deglycosylated sample and verify ASIC1 identity |
| Band lower than expected | An isoform or protein fragment is possible, but no lower mass is established | Check antibody epitope coverage and compare with an ASIC1 depletion control |
| Multiple bands | Named ASIC1 isoforms could differ in size; additional bands could also be nonspecific | Compare isoform expression and test which bands diminish with ASIC1 depletion |
| Weak or no signal | Limited recovery of multi-pass membrane ASIC1 may reduce signal | Check extraction conditions and use a membrane-enriched sample |
Experiment evidence
Western blot images showing ASIC1 band
A02332-2 · ASIC1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ASIC1 Western blot
🧪For positive controls for ASIC1 in Western blot, you can use an HPA-supported sample once identified; none is provided in the supplied evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No HPA tissue controls can be validated from the supplied data, and ASIC1 is a membrane protein.
HPA tissue expression evidence for ASIC1
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced ASIC1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ASIC1, answered from its protein features.
How should ASIC1 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 ASIC1 isoforms produce different bands?
Isoforms · Three isoforms are listed: Asic1a, 1, and Asic1b. Asic1b replaces residues 1–185, while isoform 1 changes the sequence at residue 433. Consider which isoforms the antibody recognizes when assigning bands; their observed positions are not supplied.
How can glycosylation affect ASIC1 band interpretation?
PTM · ASIC1 has N-linked glycosylation sites at UniProt Asn368 and Asn395. Compare treated and untreated samples if testing whether glycans affect mobility; the sites alone cannot identify the cause of an unexpected band.
UniProt lists phosphoserine at residue 479, modified by PKA, and phosphoserine at residue 499. Use these UniProt coordinates when discussing sites; paper or antibody numbering may differ. The features do not establish a detectable band shift.
UniProt lists phosphoserine at residue 479, modified by PKA, and phosphoserine at residue 499. Use these UniProt coordinates when discussing sites; paper or antibody numbering may differ. The features do not establish a detectable band shift.
Does this guide establish induction of ASIC1?
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 ASIC1 Western blot?
Transfer · ASIC1 is a multi-pass cell-membrane protein. Check transfer of the approximately 60 kDa region by inspecting the membrane and post-transfer gel, then optimize conditions for your setup. The supplied features do not establish a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02332-2 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 ASIC1 bands be quantified?
Quantitation · Quantify the same assigned ASIC1 band across samples using exposures within the detection range and a consistent loading reference. If multiple bands appear, consider the three listed isoforms and the glycosylation sites before combining their signals.
Why might ASIC1 migrate near 60 kDa?
Interpretation · The supplied predicted mass is 59.9 kDa, and the observed band is approximately 60 kDa. N-linked glycosylation at UniProt Asn368 and Asn395 could affect migration, but the listed sites alone do not establish a visible shift.
ASIC1 forms functional homotrimeric channels and can form heterotrimers with other ASIC proteins. A higher Western blot band alone does not establish that a trimer survived sample preparation; confirm band identity before assigning it to a channel complex.
ASIC1 forms functional homotrimeric channels and can form heterotrimers with other ASIC proteins. A higher Western blot band alone does not establish that a trimer survived sample preparation; confirm band identity before assigning it to a channel complex.
Boster reagents
ASIC1 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-ASIC1 Antibody Picoband®
Cat # A02332-2
The catalog reports one anti-ASIC1 antibody for Western blot, A02332-2. Its product image reports an approximately 60 kDa band in human THP-1 whole-cell lysate under reducing conditions, matching the expected size. The supplied evidence is limited to this reported blot.
Which to pick: A02332-2 is the only listed option. Its stated reactivity is human, and its WB image uses human THP-1 lysate (50 μg per lane; primary antibody 0.5 μg/mL). Compare that tested context with your planned sample.
Source: BosterBio ASIC1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P78348.
- Human Protein Atlas. ASIC1 tissue expression.
- PMC4542551 — target-verified WB comparison
- PMC3998777 — target-verified WB comparison
- PMC11940220 — target-verified WB comparison