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- Table of Contents
IFNAR2 / Interferon alpha/beta receptor 2 · Western blot design guide
Design a Western Blot for IFNAR2
Real validated IFNAR2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-IFNAR2 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
IFNAR2 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 | ~57.8 kDa | |
| Observed band | ~65 kDa | |
| Gel | 5–20% (catalog A02056-3) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated IFNAR2 Western Blot Protocols
The A02056-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human Caco-2, human HepG2 (catalog A02056-3) |
| Gel % | 5–20% (catalog A02056-3) |
| Load | 50ug; reducing conditions (catalog A02056-3) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A02056-3) |
| Membrane | Nitrocellulose membrane (catalog A02056-3) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A02056-3) |
| Primary antibody | A02056-3 · 0.5 μg/mL (catalog A02056-3) |
| Primary incubation | overnight at 4°C (catalog A02056-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:10000 (catalog A02056-3) |
| Secondary incubation | 1.5 hour at RT (catalog A02056-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02056-3) |
| Detection | ECL (catalog A02056-3) |
Section 2
What Is the Expected IFNAR2 Western Blot Band Size?
IFNAR2 has a predicted full-length mass of 57.8 kDa and an empirical band at ~65 kDa; the cause of their difference is not established.
| Band at ~65 kDa | Empirical IFNAR2 signal in reducing whole-cell blots; confirm identity with controls |
| Band near 57.8 kDa | Migration near the predicted full-length precursor mass; confirm identity |
| Band below the precursor position | May reflect signal-peptide cleavage or a shorter isoform; identity requires confirmation |
| Several bands | Could reflect IFNAR2 isoforms 1, 2, and 3; distinct migration is not established |
| Broad band or smear | Could reflect variable N-linked glycosylation; confirm the cause experimentally |
💡Expected IFNAR2 appearanceUniProt predicts a 57.8 kDa full-length precursor, while reducing whole-cell blots show ~65 kDa; the cause of the difference is unproven, so confirm band identity with controls.
| Predicted full-length mass | 57.8 kDa is the sequence-based reference, not a measured migration position |
| N-linked sites at Asn58, Asn87, Asn116, Asn188, and Asn192 | Glycosylation may raise or broaden apparent migration; its contribution to the ~65 kDa band is unproven |
| Signal peptide at residues 1–26 | Cleavage makes the mature chain smaller than the full-length precursor; its band position is not supplied |
| Splice isoforms 1, 2, and 3 | Their lengths may differ, but separate band positions are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | N-linked glycosylation may affect migration, but the cause of the ~65 kDa position is unproven | Compare with the empirical ~65 kDa band and confirm identity with an independent antibody or IFNAR2 depletion |
| Band lower than expected | Signal-peptide cleavage or an isoform is possible | Check antibody epitope coverage and confirm the band by IFNAR2 depletion |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible | Compare treated and untreated samples with a deglycosylation control |
| Multiple bands | Isoforms 1, 2, and 3 or nonspecific binding are possible | Identify responsive bands by IFNAR2 depletion and an independent antibody |
| Weak or no signal | Membrane IFNAR2 may be poorly recovered or expressed at low levels | Check membrane-protein recovery, loading, and a positive control |
Experiment evidence
Western blot images showing IFNAR2 band
A02056-3 · IFNAR2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for IFNAR2 Western blot
🧪For positive controls for IFNAR2 in Western blot, you can use a verified IFNAR2-expressing cell lysate, since the supplied HPA data identify no positive tissue or cell line.
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 such as Ponceau alongside the samples.
⚠️Feasibility: With no HPA expression data, positive samples need verification and no clean negative tissue can be selected.
HPA tissue expression evidence for IFNAR2
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 IFNAR2 Western Blot Tips
Deeper troubleshooting and optimisation questions for IFNAR2, answered from its protein features.
How should IFNAR2 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.
How could IFNAR2 isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Relative to the canonical sequence, isoform 2 replaces residues 281..331 and lacks 332..515; isoform 3 changes 238..239 and lacks 240..515. Check the antibody epitope before assigning a smaller band to either isoform. These coordinates use canonical UniProt numbering.
Which IFNAR2 glycosylation sites should I consider?
PTM · UniProt lists N-linked sites at Asn58, Asn87, Asn116, Asn188 and Asn192. These are canonical UniProt coordinates; antibody or paper numbering may differ. A glycosidase comparison could test whether glycosylation contributes to an apparent band shift.
The listed sites are phosphotyrosine 337 and 512, and phosphoserine 400 and 467, in canonical UniProt coordinates. Check which site the antibody recognizes and whether its epitope is present in the isoform studied. The listed sequence changes remove these canonical positions from isoforms 2 and 3; site presence alone does not establish a visible shift.
The listed sites are phosphotyrosine 337 and 512, and phosphoserine 400 and 467, in canonical UniProt coordinates. Check which site the antibody recognizes and whether its epitope is present in the isoform studied. The listed sequence changes remove these canonical positions from isoforms 2 and 3; site presence alone does not establish a visible shift.
Does this guide establish induction of IFNAR2?
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 IFNAR2 Western blot?
Transfer · Isoform 1 is a single-pass cell-membrane protein, and the reported band is about 65 kDa. Use a transfer workflow validated for membrane proteins and verify transfer around 65 kDa. The supplied features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02056-3 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 IFNAR2 bands across samples?
Quantitation · Quantify the same band or defined isoform bands consistently, and verify that the antibody detects the isoforms being compared. IFNAR2 has three isoforms, five listed N-linked sites and four listed phosphorylation sites, so a change in band pattern alone does not establish a change in total IFNAR2 abundance.
Why might IFNAR2 appear near 65 kDa instead of 57.8 kDa?
Interpretation · 57.8 kDa is the predicted mass; the observed band is about 65 kDa. IFNAR2 has five listed N-linked glycosylation sites and a signal peptide at residues 1..26. These features warrant checking processing and glycosylation, but their presence alone does not establish the cause or size of the difference.
Check whether the antibody epitope is retained in each isoform, especially given the missing C-terminal regions in isoforms 2 and 3. Consider the five listed N-linked sites and the 1..26 signal peptide when evaluating apparent size. Assigning a band to an isoform or modification requires evidence beyond these feature annotations.
Check whether the antibody epitope is retained in each isoform, especially given the missing C-terminal regions in isoforms 2 and 3. Consider the five listed N-linked sites and the 1..26 signal peptide when evaluating apparent size. Assigning a band to an isoform or modification requires evidence beyond these feature annotations.
Boster reagents
IFNAR2 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-IFNAR2 Antibody Picoband®
Cat # A02056-3
A02056-3 is listed for human, mouse, and rat IFNAR2. Its Western blot image shows an approximately 65 kDa band in human K562, Caco-2, HepG2, and U-87MG whole-cell lysates. The supplied image does not show mouse or rat samples.
Which to pick: A02056-3 is the only listed option and has a Western blot image using human cell lysates. For mouse or rat samples, its listed reactivity is available, but the supplied image provides no Western blot example in those species.
Source: BosterBio IFNAR2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P48551.
- Human Protein Atlas. IFNAR2 tissue expression.
- PMC3188543 — target-verified WB comparison
- PMC6795910 — target-verified WB comparison