ILF2 / Interleukin enhancer-binding factor 2 · Western blot design guide

Design a Western Blot for ILF2

Source-linked ILF2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ILF2 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

Western blot protocol sheet for ILF2: expected band ~43.1 kDa, hero antibody A04443-2, catalog values and labelled standard workflow; separate PMC comparisons on the guide — Printable **ILF2** Western blot protocol sheet — expected band ~43.1 kDa, antibody A04443-2, controls and PMC citations. Open the full ILF2 WB guide →

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ILF2 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	~43.1 kDa
	Observed band	~43 kDa
	Gel	5–20% (catalog A04443-2)
	Positive control ⓘ	Adipose tissue (IHC candidate; verify WB) +4 more
	Negative control ⓘ	Suggested KO / knockdown lysate

Important caveats

Reasons your observed band may differ from the expected size.

	PTM	Phosphorylated + Methylated
	Caveat	Phosphorylation-state controls
	Gene-set association	MSigDB Hallmark membership
	Isoform	1 isoform(s)

Section 1

Source-Linked ILF2 Western Blot Protocol Options

The A04443-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.

Recommended Western blot protocol parameters
Sample / lysate	human K562, human Jurkat, human Hela, monkey COS-7, rat brain, rat C6, mouse brain, mouse NIH/3T3 (catalog A04443-2)
Gel %	5–20% (catalog A04443-2)
Load	30 ug; reducing conditions (catalog A04443-2)
Transfer	a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04443-2)
Membrane	nitrocellulose membrane (catalog A04443-2)
Blocking	5% non-fat milk/TBS for 1.5 hour at RT (catalog A04443-2)
Primary antibody	A04443-2 · 0.25 μg/mL (catalog A04443-2)
Primary incubation	overnight at 4°C (catalog A04443-2)
Secondary antibody	goat anti-rabbit IgG-HRP, 1:5000 (catalog A04443-2)
Secondary incubation	1.5 hour at RT (catalog A04443-2)
Wash	TBS-0.1%Tween 3 times with 5 minutes each (catalog A04443-2)
Detection	ECL (catalog A04443-2)

Section 2

What Is the Expected ILF2 Western Blot Band Size?

ILF2 is predicted at 43.1 kDa and observed at ~43 kDa; the small difference has no established cause in the supplied evidence.

What am I looking at on my blot?
Band at ~43 kDa in whole-cell lysate	Consistent with ILF2; confirm identity with antibody controls
Band at ~43 kDa in a nuclear fraction	Consistent with ILF2 nuclear localization
Band at ~43 kDa in a cytoplasmic fraction	Consistent with ILF2 cytoplasmic localization
Close bands near ~43 kDa	Could reflect modified ILF2, but the listed sites do not establish a visible doublet

💡Expected ILF2 appearanceILF2 is predicted at 43.1 kDa and observed at ~43 kDa in reducing whole-cell blots; confirm band identity with appropriate antibody controls.

How each factor affects band size
Predicted ILF2 mass	43.1 kDa, consistent with the observed ~43 kDa band
Arg16 and Arg24 methylation	Adds little mass; a visible migration change is not established
Ser52 and Ser68 phosphorylation	May alter migration, but a distinct band is not established
Thr388 phosphorylation	May alter migration, but a distinct band is not established

Why is my band missing or off?
Situation	Likely cause	Next action
No band in lysate	ILF2 recovery or detection may be inadequate	Check loading and antibody performance against a whole-cell lysate with a reported ~43 kDa band
Band higher than expected	Its identity is uncertain; listed phosphorylation sites do not establish a large shift	Confirm specificity with an independent antibody or ILF2 depletion
Band lower than expected	Its identity is uncertain; no cleavage feature is listed	Check sample integrity and confirm specificity by ILF2 depletion
Multiple bands	Modified ILF2 or nonspecific antibody binding is possible	Compare bands after ILF2 depletion; assess phosphorylation if relevant
Weak or no signal	Extraction may recover nuclear or cytoplasmic ILF2 poorly	Check lysate preparation and use a whole-cell positive control
Fragments below expected size	Sample degradation is possible	Prepare fresh lysate with protease inhibitors and compare with a positive control

Source: UniProt Q12905 Human Protein Atlas product datasheet (A04443-2)

Experiment evidence

Western blot images showing ILF2 band

A04443-2 · ILF2

M04443-1 · ILF2

← swipe / scroll for more images →

Real WB data behind the band-size expectations in this section.

Sample controls for ILF2 Western blot

🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ILF2 in Western blot, you can use adipose tissue, which HPA rates as highly expressed.

Positive control: Adipose tissue (IHC candidate; verify WB)

Negative control: Suggested KO / knockdown lysate

Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.

⚠️Feasibility: No Not-detected tissue is supplied, so use siRNA knockdown or a KO line as the negative control.

HPA tissue expression evidence for ILF2

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
Adipose tissue	adipocytes	High	Protein (IHC)	HPA →
Adrenal gland	glandular cells	High	Protein (IHC)	HPA →
Appendix	glandular cells	High	Protein (IHC)	HPA →
Bone marrow	hematopoietic cells	High	Protein (IHC)	HPA →
Breast	adipocytes	High	Protein (IHC)	HPA →

Lower expression tissues · IHC evidence, not confirmed WB-negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	adipocytes	High	Protein (IHC)	HPA →
Adrenal gland	glandular cells	High	Protein (IHC)	HPA →
Appendix	glandular cells	High	Protein (IHC)	HPA →
Bone marrow	hematopoietic cells	High	Protein (IHC)	HPA →
Breast	adipocytes	High	Protein (IHC)	HPA →

Section 3

Advanced ILF2 Western Blot Tips

Deeper troubleshooting and optimisation questions for ILF2, answered from its protein features.

How should ILF2 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 annotated ILF2 isoforms explain multiple bands?

Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no annotated isoform explanation for multiple bands.

Which phosphorylation sites should I consider when interpreting ILF2 bands?

PTM · UniProt lists phosphoserine at positions 52 and 68 and phosphothreonine at 388. These are UniProt sequence coordinates; antibody or paper numbering may differ. Their presence alone does not predict a distinct band.

Does this guide establish induction of ILF2?

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 ILF2?

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 A04443-2 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 ILF2 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.

How does the observed ILF2 band compare with predicted mass?

Interpretation · The observed band near 43 kDa agrees with the predicted 43.1 kDa. Listed modifications do not, by themselves, establish a visible shift.

UniProt lists arginine 16 as alternatively asymmetric dimethylated or omega-N-methylated, and arginine 24 as omega-N-methylated. The alternatives at position 16 should not be counted as simultaneous modifications. These are UniProt sequence coordinates.

ILF2 is listed in the nucleus, nucleolus and cytoplasm, including cytoplasmic mRNP granules. Keep fraction choice consistent when comparing samples; a change in one fraction need not represent a change in total ILF2.

The record lists methylation, phosphorylation and association with ILF3, but none establishes the identity of an unexpected band. Compare it with the approximately 43 kDa band and validate its identity before assigning it to a modification or complex.

Boster reagents

ILF2 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

Western blot analysis of NF45/ILF2 using anti-NF45/ILF2 antibody (A04443-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human K562 whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: monkey COS-7 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat C6 whole cell lysates, Lane 7: mouse brain tissue lysates, Lane 8: mouse NIH/3T3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NF45/ILF2 antigen affinity purified polyclonal antibody (Catalog # A04443-2) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for NF45/ILF2 at approximately 43 kDa. The expected band size for NF45/ILF2 is at 43 kDa.

Anti-NF45/ILF2 Antibody Picoband®

Cat # A04443-2

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Real WB data

Western blot analysis of ILF2 using anti-ILF2 antibody (M04443-1). Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human A431 whole cell lysates, Lane 2: human PC-3 whole cell lysates, Lane 3: human SH-SY5Y whole cell lysates, Lane 4: human K562 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: mouse brain tissue lysates, Lane 7: mouse kidney tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ILF2 antigen affinity purified monoclonal antibody (M04443-1) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for ILF2 at approximately 43 kDa. The expected band size for ILF2 is at 43 kDa.

Anti-ILF2 Rabbit Monoclonal Antibody

Cat # M04443-1

View product →

Both listed anti-ILF2 antibodies have product Western blot images showing an approximately 43 kDa band in the tested human, mouse, and rat lysates; A04443-2 also shows monkey COS-7 lysate. These are product examples, with no independent validation supplied.

Which to pick: For monkey COS-7 lysate, choose A04443-2, which has a matching blot lane. For human, mouse, or rat samples, compare the pictured lysates with your sample: both antibodies have WB images, but their tested specimens differ.

Source: BosterBio ILF2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.

References

UniProt Consortium. UniProt entry Q12905.
Human Protein Atlas. ILF2 tissue expression.
PMC6942249 — target-verified WB comparison
PMC6936233 — target-verified WB comparison
PMC5360436 — target-verified WB comparison