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- Table of Contents
IFIT1 / Antiviral innate immune response effector IFIT1 · Western blot design guide
Design a Western Blot for IFIT1
Real validated IFIT1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-IFIT1 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
IFIT1 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 | ~55.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebral cortex (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 + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated IFIT1 Western Blot Protocols
The A02652-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat small intestine tissue lysate (catalog A02652-1) |
| 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 | A02652-1 · 1 μg/ml (catalog A02652-1) |
| 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 IFIT1 Western Blot Band Size?
IFIT1 has a predicted mass of 55.4 kDa; alternative splicing could affect migration, but no empirical band size or resolvable isoform pattern is supplied.
| Single band near 55.4 kDa | consistent with predicted IFIT1 size; confirm identity with a control |
| Additional band above 55.4 kDa | could reflect an IFIT1 isoform; its size is unknown |
| Additional band below 55.4 kDa | could reflect an IFIT1 isoform; its size is unknown |
| Several bands at different sizes | isoforms 1 and 2 could contribute if they migrate differently; confirm band identities |
💡Expected IFIT1 appearanceUniProt predicts 55.4 kDa for IFIT1; no empirical band size or isoform migration is supplied, so use a positive control and IFIT1 depletion to identify the band.
| Predicted IFIT1 mass | 55.4 kDa provides the reference size, not a measured migration |
| Isoform 1 | may differ in size from isoform 2; its mass is not supplied |
| Isoform 2 | may differ in size from isoform 1; its mass is not supplied |
| Alternative splicing | could produce different apparent sizes, but separation is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | IFIT1 may be below detection in the tested sample | check a positive control and increase lysate input |
| Band higher than expected | band identity or migration is unestablished | compare with a positive control and IFIT1 depletion |
| Band lower than expected | an alternate isoform is possible, but its size is unknown | check isoform coverage and confirm with IFIT1 depletion |
| Multiple bands | isoforms 1 and 2 are listed, but their separation is unknown | confirm which bands depend on IFIT1 |
| Weak or no signal | IFIT1 may be below detection or antibody detection may be insufficient | check a positive control and antibody conditions |
| Fragments below expected size | sample degradation is possible; no cleavage feature is listed | prepare fresh lysate with protease inhibitors and confirm IFIT1 dependence |
Experiment evidence
Western blot images showing IFIT1 band
A02652-1 · IFIT1
A02652 · IFIT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for IFIT1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for IFIT1 in Western blot, you can use cerebral cortex lysate, which HPA scores High for IFIT1.
Positive control: Cerebral cortex (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: Cytoplasmic localization supports using tissue lysates, and HPA identifies adipose tissue as a candidate negative control.
HPA tissue expression evidence for IFIT1
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 |
|---|---|---|---|---|
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IFIT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for IFIT1, answered from its protein features.
Where should the main IFIT1 band appear?
Band shift · The canonical sequence has a predicted mass of 55.4 kDa. No empirical apparent band position is supplied, so use 55.4 kDa as a starting point rather than an exact migration target.
Could IFIT1 isoforms produce different bands?
Isoforms · Two isoforms are listed. Isoform 2 lacks residues 1–31 in UniProt canonical numbering, so it may migrate differently from isoform 1. The supplied features do not establish whether both isoforms are present in your sample or resolved on your blot.
Do IFIT1 modifications explain a shifted band?
PTM · IFIT1 is tagged as a phosphoprotein and as subject to Ubl conjugation, but no modified residues or glycosylation sites are listed. These features alone do not establish a visible shift or explain a difference from the predicted mass.
Does this guide establish induction of IFIT1?
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 IFIT1?
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 A02652-1 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 IFIT1 bands be quantified?
Quantitation · Define the band or bands being measured before comparing samples. Two isoforms are listed, and isoform 2 lacks canonical residues 1–31; if distinct bands appear, quantify them consistently and avoid assigning either band to an isoform from position alone.
How should interferon conditions be compared?
Interpretation · IFIT1 is a component of an interferon-dependent complex with IFIT2 and IFIT3. Compare samples collected under defined, matched interferon conditions. That complex annotation alone does not establish how much IFIT1 protein changes after treatment.
IFIT1 is annotated in the cytoplasm. A cytoplasmic fraction is therefore a relevant sample for checking its band; record the fraction used when comparing results.
Check their positions against the 55.4 kDa canonical prediction and the documented N-terminal deletion in isoform 2. Neither an empirical band position nor a modification-specific shift is supplied, so band position alone cannot establish an isoform or modification.
IFIT1 is annotated in the cytoplasm. A cytoplasmic fraction is therefore a relevant sample for checking its band; record the fraction used when comparing results.
Check their positions against the 55.4 kDa canonical prediction and the documented N-terminal deletion in isoform 2. Neither an empirical band position nor a modification-specific shift is supplied, so band position alone cannot establish an isoform or modification.
Boster reagents
IFIT1 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-IFIT1 Antibody
Cat # A02652-1
Real WB data
Anti-IFIT1 Antibody
Cat # A02652
Two anti-IFIT1 antibodies are listed for Western blot, each with stated human, mouse, and rat reactivity and a WB image. The supplied images show specific sample contexts; they do not establish performance across every listed species.
Which to pick: Choose A02652-1 for its rat small intestine lysate example at 1 μg/ml, or A02652 for its LOVO cell lysate example with a peptide-blocked lane. Both have WB images; select the example closest to your planned sample and conditions.
Source: BosterBio IFIT1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P09914.
- Human Protein Atlas. IFIT1 tissue expression.
- PMC9664554 — target-verified WB comparison
- PMC5850363 — target-verified WB comparison
- PMC8973581 — target-verified WB comparison