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EIF3H / Eukaryotic translation initiation factor 3 subunit H · Western blot design guide
Design a Western Blot for EIF3H
Source-linked EIF3H Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-EIF3H 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
EIF3H 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 | ~39.9 kDa | |
| Observed band | ~40 kDa | |
| Gel | 10% (catalog A05656-1) | |
| Positive control | Appendix (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 + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked EIF3H Western Blot Protocol Options
The A05656-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEL, human K562, human THP-1, human PC-3, rat brain, mouse brain (catalog A05656-1) |
| Gel % | 10% (catalog A05656-1) |
| Load | 30 ug; reducing conditions (catalog A05656-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05656-1) |
| Membrane | nitrocellulose membrane (catalog A05656-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05656-1) |
| Primary antibody | A05656-1 · 0.5 μg/mL (catalog A05656-1) |
| Primary incubation | overnight at 4°C (catalog A05656-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05656-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05656-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05656-1) |
| Detection | ECL (catalog A05656-1) |
Section 2
What Is the Expected EIF3H Western Blot Band Size?
EIF3H is predicted at 39.9 kDa and observed near 40 kDa; the small difference has no established cause in the supplied evidence.
| Band near 40 kDa | Matches the empirical EIF3H band and its 39.9 kDa predicted mass |
| Faint band near 40 kDa | Could reflect low detectable EIF3H in the lysate |
| No band near 40 kDa | EIF3H was not detected in this cytoplasmic sample |
| Close doublet near 40 kDa | Could reflect different phosphorylation states, but a mobility effect is unproven |
💡Expected EIF3H appearanceEIF3H has a predicted mass of 39.9 kDa and an empirical band near 40 kDa; use antibody and sample controls to confirm band identity.
| Predicted molecular mass | Places the unmodified 352-residue protein near 39.9 kDa |
| 352-residue sequence | Defines the full-length protein underlying the predicted mass |
| Phosphoserine at Ser3 | May affect mobility, but no visible shift is established |
| Phosphoserine at Ser183 | May affect mobility, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | EIF3H is not detected despite its cytoplasmic location | Check lysate loading and a positive-control lysate |
| Band higher than expected | Band identity or altered mobility is uncertain | Compare with the 40 kDa reference and confirm antibody specificity |
| Band lower than expected | Possible breakdown or an unrelated recognized protein | Check sample integrity and antibody specificity |
| Multiple bands | Additional bands are not explained by listed isoforms | Compare with a positive control and validate the 40 kDa band |
| Weak or no signal | Insufficient detectable EIF3H or assay sensitivity | Check protein loading and antibody detection with a positive control |
| Fragments below expected size | Possible sample breakdown | Prepare fresh lysate with protease inhibitors and compare with the 40 kDa band |
Experiment evidence
Western blot images showing EIF3H band
A05656-1 · EIF3H
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for EIF3H Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for EIF3H in Western blot, you can use appendix tissue lysate.
Positive control: Appendix (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 tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for EIF3H
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced EIF3H Western Blot Tips
Deeper troubleshooting and optimisation questions for EIF3H, answered from its protein features.
How should EIF3H 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.
Do EIF3H isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no alternative isoform sequence to assign to another band.
Could phosphorylation shift the EIF3H band?
PTM · UniProt lists phosphoserine at positions 3 and 183. These coordinates use UniProt numbering, which may differ from paper or antibody numbering. Site annotation alone does not show that phosphorylation produces a visible band shift.
Does this guide establish induction of EIF3H?
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 EIF3H Western blot?
Transfer · EIF3H is predicted at 39.9 kDa. Choose transfer conditions that recover proteins around 40 kDa, and verify transfer in that size range. The supplied features do not specify a membrane or transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05656-1 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 EIF3H be quantified?
Quantitation · Quantify the band near the observed 40 kDa position consistently across samples. If additional bands appear, do not combine them with the main band solely because EIF3H has annotated phosphorylation sites.
Why is the EIF3H band near 40 kDa?
Interpretation · EIF3H has a predicted mass of 39.9 kDa, consistent with the observed band near 40 kDa. Its listed features do not establish a separate cause for any small difference between predicted and apparent mass.
Use the observed band near 40 kDa as the reference. The listed phosphorylation sites do not by themselves identify another band, and the record provides no alternative sequence or glycosylation site to assign one.
UniProt locates EIF3H in the cytoplasm and identifies it as a component of the eIF-3 complex. A cytoplasmic sample is therefore relevant when checking for its band near 40 kDa.
Use the observed band near 40 kDa as the reference. The listed phosphorylation sites do not by themselves identify another band, and the record provides no alternative sequence or glycosylation site to assign one.
UniProt locates EIF3H in the cytoplasm and identifies it as a component of the eIF-3 complex. A cytoplasmic sample is therefore relevant when checking for its band near 40 kDa.
Boster reagents
EIF3H 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-EIF3H Antibody Picoband®
Cat # A05656-1
The catalog reports one anti-EIF3H antibody for Western blot, A05656-1. Its WB image reports an approximately 40 kDa band, matching the expected size, in human cell lysates and rat and mouse brain lysates. This is product-reported evidence; no independent validation is supplied.
Which to pick: A05656-1 is the only listed option. It is reported reactive with human, mouse, and rat, with a WB image showing human HEL, K562, THP-1, and PC-3 cells plus rat and mouse brain lysates under the stated conditions.
Source: BosterBio EIF3H gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O15372.
- Human Protein Atlas. EIF3H tissue expression.
- PMC13564525 — target-verified WB comparison
- PMC5952081 — target-verified WB comparison
- PMC10413709 — target-verified WB comparison