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- Table of Contents
IFIT2 / Interferon-induced protein with tetratricopeptide repeats 2 · Western blot design guide
Design a Western Blot for IFIT2
Real validated IFIT2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-IFIT2 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
IFIT2 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 | ~54.6 kDa | |
| Observed band | ~55 kDa | |
| Gel | 10% (catalog A04428-1) | |
| Positive control | Kidney (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Interferon-induction controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated IFIT2 Western Blot Protocols
The A04428-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hacat, human A431, human SIHA (catalog A04428-1) |
| Gel % | 10% (catalog A04428-1) |
| Load | 30 ug; reducing conditions (catalog A04428-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04428-1) |
| Membrane | nitrocellulose membrane (catalog A04428-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04428-1) |
| Primary antibody | A04428-1 · 0.5 μg/mL (catalog A04428-1) |
| Primary incubation | overnight at 4°C (catalog A04428-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04428-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04428-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04428-1) |
| Detection | ECL (catalog A04428-1) |
Section 2
What Is the Expected IFIT2 Western Blot Band Size?
IFIT2 is predicted at 54.6 kDa and observed near 55 kDa; the cause of their small difference is not established.
| Band near 55 kDa | Consistent with the observed IFIT2 band and its 54.6 kDa predicted mass |
| Band near 110 kDa | Could reflect a domain-swapped homodimer if it survives sample preparation |
| Higher-mass band under mild preparation | Could reflect an IFIT1–IFIT2–IFIT3 complex if it remains intact |
| Weak or absent band | May reflect low IFIT2 abundance without interferon induction |
💡Expected IFIT2 appearanceIFIT2 has a predicted mass of 54.6 kDa and an observed band near 55 kDa; confirm band identity with appropriate antibody and expression controls.
| Predicted IFIT2 mass | Sets the 54.6 kDa monomer reference, consistent with the observed ~55 kDa band |
| Ser2 N-acetylation | Is documented, but no visible migration effect is established |
| Domain-swapped homodimer | Could appear near twice the monomer mass if retained during preparation |
| IFIT1–IFIT2–IFIT3 complex | Could produce a higher-mass species if retained; its migration is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | IFIT2 abundance may be low without interferon induction | Compare induced and uninduced lysates with a loading control |
| Band higher than expected | A domain-swapped homodimer may persist during preparation | Compare fully denatured and milder preparations and verify band identity |
| Band lower than expected | The lower band is not explained by a listed cleavage feature | Check sample integrity and confirm identity with an independent antibody |
| Multiple bands | Additional bands have no established isoform explanation in the supplied features | Compare induced and uninduced samples and verify specificity with IFIT2 depletion |
| Weak or no signal | IFIT2 expression may be low in the tested sample | Check loading and compare with an interferon-induced positive control |
| Fragments below expected size | Sample degradation may produce smaller immunoreactive fragments | Prepare fresh lysate with protease inhibitors and check whether the ~55 kDa band is retained |
Experiment evidence
Western blot images showing IFIT2 band
A04428-1 · IFIT2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for IFIT2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for IFIT2 in Western blot, you can use kidney tissue, which has high HPA expression.
Positive control: Kidney (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Kidney and adipose tissue offer feasible tissue controls, but their Western blot signals need validation.
HPA tissue expression evidence for IFIT2
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 |
|---|---|---|---|---|
| Kidney | cells in tubules | High | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IFIT2 Western Blot Tips
Deeper troubleshooting and optimisation questions for IFIT2, answered from its protein features.
How should IFIT2 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.
Are multiple IFIT2 bands expected from annotated isoforms?
Isoforms · Only one isoform is supplied, with no alternative sequence. Do not assign additional bands to annotated IFIT2 isoforms on this evidence alone.
Could N-terminal acetylation shift the IFIT2 band?
PTM · UniProt lists N-acetylserine at position 2, using UniProt sequence numbering. Its presence does not establish a visible band shift; compare the apparent band with the reported ~55 kDa position.
Does this guide establish induction of IFIT2?
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 IFIT2?
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 A04428-1 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 IFIT2 band intensity be quantified?
Quantitation · Measure the band near 55 kDa across samples prepared and analyzed consistently. Because IFIT2 is interferon-induced and occurs in cytoplasm and endoplasmic reticulum, account for treatment and sample fraction when comparing intensities.
How does IFIT2's observed band compare with its predicted mass?
Interpretation · The reported band near 55 kDa closely matches IFIT2's predicted 54.6 kDa. The listed modification alone does not establish a visible shift or explain any mass difference.
IFIT2 is interferon-induced and belongs to an interferon-dependent complex. Comparing matched untreated and interferon-exposed samples can help assess the response, but these features do not specify a dose, time course, or expected fold change.
IFIT2 forms a domain-swapped homodimer and interacts with IFIT1 and IFIT3. These features make complexes relevant to investigate, but they do not establish that an unexpected band contains IFIT2 or that it survives the blot conditions.
IFIT2 is reported in the cytoplasm and endoplasmic reticulum. Record whether samples are whole-cell lysates or fractions, and compare like with like when interpreting band intensity.
IFIT2 is interferon-induced and belongs to an interferon-dependent complex. Comparing matched untreated and interferon-exposed samples can help assess the response, but these features do not specify a dose, time course, or expected fold change.
IFIT2 forms a domain-swapped homodimer and interacts with IFIT1 and IFIT3. These features make complexes relevant to investigate, but they do not establish that an unexpected band contains IFIT2 or that it survives the blot conditions.
IFIT2 is reported in the cytoplasm and endoplasmic reticulum. Record whether samples are whole-cell lysates or fractions, and compare like with like when interpreting band intensity.
Boster reagents
IFIT2 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-IFIT2 Antibody Picoband®
Cat # A04428-1
The catalog reports one anti-IFIT2 antibody for WB, A04428-1, with stated human, mouse, and rat reactivity. Its product WB image reports a band at approximately 55 kDa in the specified lysates. No independent publication or orthogonal validation evidence is supplied.
Which to pick: A04428-1 is the only listed option. Its WB image includes human Hacat, A431, SIHA, and PC-3 cell lysates; rat liver and RH35 lysates; and mouse liver and HEPA1-6 lysates. Check whether those tested samples and the stated reactivity fit your experiment.
Source: BosterBio IFIT2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P09913.
- Human Protein Atlas. IFIT2 tissue expression.
- PMC7761248 — target-verified WB comparison
- PMC11253605 — target-verified WB comparison
- PMC3355090 — target-verified WB comparison