This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
HIP1 / Huntingtin-interacting protein 1 · Western blot design guide
Design a Western Blot for HIP1
Source-linked HIP1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HIP1 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
HIP1 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 | ~116.2 kDa | |
| Observed band | Approximately 116 kDa | |
| Gel | 5–20% (catalog M02242) | |
| Positive control | Caudate (IHC candidate; verify WB) +4 more | |
| Negative control | Salivary gland (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked HIP1 Western Blot Protocol Options
The M02242 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human MCF-7, human RT4, human SH-SY5Y, rat brain, mouse brain (catalog M02242) |
| Gel % | 5–20% (catalog M02242) |
| Load | 30 ug; reducing conditions (catalog M02242) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02242) |
| Membrane | nitrocellulose membrane (catalog M02242) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M02242) |
| Primary antibody | M02242; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M02242) |
| Secondary incubation | 1.5 hour at RT (catalog M02242) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M02242) |
| Detection | ECL (catalog M02242) |
Section 2
What Is the Expected HIP1 Western Blot Band Size?
HIP1 is predicted at 116.2 kDa and observed at approximately 116 kDa; phosphorylation, homodimerization and splice isoforms have no demonstrated migration effect here.
| Band at approximately 116 kDa | Matches the empirical HIP1 band and the predicted 116.2 kDa mass; confirm identity with antibody controls. |
| Band near twice the monomer size | Could reflect an SDS-resistant HIP1 homodimer; dimer migration has not been demonstrated here. |
| Several bands near the expected region | Could include HIP1 splice isoforms, but distinct migration has not been established. |
| Band below 116 kDa | Could be a smaller HIP1 isoform; its identity requires verification. |
💡Expected HIP1 appearanceHIP1 has a predicted mass of 116.2 kDa and an empirical band at approximately 116 kDa; confirm band identity with antibody controls.
| UniProt predicted mass | Places the full-length reference protein near 116.2 kDa, consistent with the observed approximately 116 kDa band. |
| Phosphoserine at residue 338 | May affect mobility, but no visible shift is established. |
| HIP1 homodimer | Could appear near twice the monomer size if it survives sample preparation and electrophoresis. |
| HIP1-1, HIP1-2, 3 and 4 splice isoforms | May differ in size, but their masses and separation on a blot are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | HIP1 may be missed if the sampled fraction excludes its cytoplasmic, nuclear or endomembrane pools. | Compare whole-cell lysate with appropriate fractions and check fraction markers. |
| Band higher than expected | A HIP1 homodimer may persist during sample preparation. | Compare fully denatured samples with the original preparation and verify band identity. |
| Band lower than expected | A smaller splice isoform is possible, but its migration is unknown. | Check antibody epitope coverage and confirm the band with an independent HIP1 antibody. |
| Multiple bands | HIP1 has four named splice isoforms, though their separation is unproven. | Compare independent HIP1 antibodies and an appropriate HIP1 depletion control. |
| Weak or no signal | HIP1 occupies multiple cellular compartments, so a selected fraction may contain little target. | Check whole-cell lysate, loading and fraction markers, then verify antibody performance with a positive control. |
Experiment evidence
Western blot images showing HIP1 band
M02242 · HIP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HIP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HIP1 in Western blot, you can use caudate tissue, where HPA reports high expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Salivary gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HIP1 shuttles between cytoplasm and nucleus, so signal may vary between cell fractions.
HPA tissue expression evidence for HIP1
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 |
|---|---|---|---|---|
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced HIP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for HIP1, answered from its protein features.
How should HIP1 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 HIP1 isoforms produce different bands?
Isoforms · UniProt lists four isoforms. Isoform 3 lacks canonical residues 803–853, while isoform 4 replaces canonical residues 1–40 with a shorter sequence. These differences could affect band position or antibody recognition; check whether the antibody epitope is retained. No sequence difference is supplied here for HIP1-2.
Does HIP1 phosphorylation explain a band shift?
PTM · UniProt lists phosphoserine at position 338. Use canonical UniProt numbering when comparing this site with an antibody or paper, which may use another convention. The listed site alone does not establish a visible shift or explain a mass difference.
Does androgen receptor activity induce HIP1 expression?
Induction · UniProt states that AR can induce HIP1 nuclear translocation. It does not establish increased HIP1 expression. If testing an AR-related condition, compare nuclear and cytoplasmic HIP1 alongside total HIP1 before interpreting a stronger nuclear band as induction.
What transfer method to use for HIP1 Western blot?
Transfer · Choose transfer conditions suitable for a protein near 116 kDa, and check transfer with a membrane stain and the post-transfer gel. The supplied features do not establish one specific transfer method or duration.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02242 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 HIP1 be quantified across cellular fractions?
Quantitation · HIP1 is found in the cytoplasm, nucleus, and endomembrane system, and shuttles between cytoplasm and nucleus. Quantify comparable fractions separately and normalize each to its appropriate loading control; a change in one fraction may reflect redistribution.
Should HIP1 run at its predicted molecular weight?
Interpretation · HIP1 is predicted at 116.2 kDa, close to the reported apparent band of approximately 116 kDa. Use that region to identify a candidate band, then confirm specificity; the mass match alone does not identify HIP1.
Check isoform sequence differences and antibody epitope coverage first. HIP1 is also listed as a homodimer and has phosphoserine 338, but those features alone do not identify an unexpected band or prove a visible shift. Compare the band with the approximately 116 kDa reference and verify antibody specificity.
Check isoform sequence differences and antibody epitope coverage first. HIP1 is also listed as a homodimer and has phosphoserine 338, but those features alone do not identify an unexpected band or prove a visible shift. Compare the band with the approximately 116 kDa reference and verify antibody specificity.
Boster reagents
HIP1 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-HIP1 Monoclonal Antibody
Cat # M02242
M02242 is listed as an anti-HIP1 monoclonal antibody reactive with human, mouse, and rat. Its WB caption reports a 116 kDa band in human cell lines and rat and mouse brain lysates, but names a different antibody code, leaving direct validation of M02242 unclear.
Which to pick: M02242 is the only listed option. Its stated reactivity covers human, mouse, and rat, but confirm that the WB image belongs to this SKU before relying on the reported blot.
Source: BosterBio HIP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00291.
- Human Protein Atlas. HIP1 tissue expression.
- PMC10753359 — target-verified WB comparison
- PMC5783582 — target-verified WB comparison
- PMC8624357 — target-verified WB comparison