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- Table of Contents
INPP4B / Inositol polyphosphate 4-phosphatase type II · Western blot design guide
Design a Western Blot for INPP4B
Source-linked INPP4B Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-INPP4B 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
INPP4B 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 | ~104.7 kDa | |
| Observed band | ~105 kDa | |
| Gel | 5–20% (catalog M06336) | |
| Positive control | Breast (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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked INPP4B Western Blot Protocol Options
The M06336 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7 (catalog M06336) |
| Gel % | 5–20% (catalog M06336) |
| Load | 30 ug; reducing conditions (catalog M06336) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M06336) |
| Membrane | nitrocellulose membrane (catalog M06336) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M06336) |
| Primary antibody | M06336 · 1:1000 (catalog M06336) |
| Primary incubation | overnight at 4°C (catalog M06336) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M06336) |
| Secondary incubation | 1.5 hour at RT (catalog M06336) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M06336) |
| Detection | ECL (catalog M06336) |
Section 2
What Is the Expected INPP4B Western Blot Band Size?
INPP4B is predicted at 104.7 kDa and observed at ~105 kDa; the cause of the small difference is not established.
| Band at ~105 kDa | Matches the reported INPP4B band; confirm identity with antibody controls |
| Single band near 104.7 kDa | Consistent with the UniProt predicted mass |
| Two bands near the expected size | Could reflect isoforms 1 and 2 if they migrate differently; confirm identity |
| No distinct isoform doublet | Isoforms 1 and 2 are not necessarily resolved on this blot |
💡Expected INPP4B appearanceUniProt predicts 104.7 kDa, and antibody QC reports a band at ~105 kDa; confirm band identity with appropriate antibody controls.
| UniProt predicted mass | 104.7 kDa is close to the observed ~105 kDa band |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its individual mass and migration are not supplied |
| Alternative splicing | May change band position if isoforms 1 and 2 differ in size; a resolvable difference is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | INPP4B may be below detection in the tested lysate | Check loading, transfer, and antibody performance with a positive control |
| Band higher than expected | Identity or migration of the higher band is unestablished | Compare with a positive control and verify antibody specificity |
| Band lower than expected | An isoform difference is possible but its direction is unknown | Verify specificity before assigning the band to an isoform |
| Multiple bands | Isoforms 1 and 2 are annotated, but their band positions are unknown | Use specificity controls to identify which bands represent INPP4B |
| Weak or no signal | INPP4B signal may be below detection | Check sample loading, transfer, and antibody performance with a positive control |
Experiment evidence
Western blot images showing INPP4B band
M06336 · INPP4B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for INPP4B Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for INPP4B in Western blot, you can use breast tissue lysate.
Positive control: Breast (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: Breast has medium HPA expression, while adipose tissue is not detected, so confirm the expected band with an additional specificity check.
HPA tissue expression evidence for INPP4B
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 |
|---|---|---|---|---|
| Breast | myoepithelial cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | 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 → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Not detected | Protein (IHC) | HPA → |
| Tonsil | germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced INPP4B Western Blot Tips
Deeper troubleshooting and optimisation questions for INPP4B, answered from its protein features.
How should INPP4B 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 isoform 2 appear at a different position?
Isoforms · Yes. In UniProt numbering, isoform 2 replaces residues 125–131 and lacks residues 132–924, so it is much shorter than the 924-residue canonical protein. Its apparent band position is not supplied; the sequence difference alone does not establish a visible band.
Check its epitope against the UniProt sequences. Residues 1–124 are shared, residues 125–131 differ, and residues 132–924 are absent from isoform 2. An antibody targeting the absent region would not be expected to detect isoform 2.
Check its epitope against the UniProt sequences. Residues 1–124 are shared, residues 125–131 differ, and residues 132–924 are absent from isoform 2. An antibody targeting the absent region would not be expected to detect isoform 2.
Does UniProt support a modification-based INPP4B band shift?
PTM · No modified residues or glycosylation sites are listed in the supplied features. Do not assign a shifted band to a specific modification from this record, or infer that a modification must cause a visible shift.
Does this guide establish induction of INPP4B?
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 INPP4B Western blot?
Transfer · For the approximately 105 kDa canonical band, verify that transfer captures protein at that size by checking the membrane and residual gel. The supplied features do not determine a specific transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M06336 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 INPP4B bands be quantified?
Quantitation · Quantify the near-105 kDa band as canonical INPP4B only after confirming antibody specificity. If measuring both isoforms, use an epitope shared by them and quantify distinguishable bands separately; the supplied features give no apparent mass for isoform 2.
Does the observed INPP4B band match its predicted mass?
Interpretation · The observed band near 105 kDa closely matches the supplied 104.7 kDa predicted mass for canonical INPP4B. A small difference does not require a modification-based explanation.
Isoform 2 is a possible explanation because it lacks UniProt residues 132–924. Check whether the antibody epitope is retained in isoform 2 before assigning the band; size alone cannot establish its identity.
Isoform 2 is a possible explanation because it lacks UniProt residues 132–924. Check whether the antibody epitope is retained in isoform 2 before assigning the band; size alone cannot establish its identity.
Boster reagents
INPP4B 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-INPP4B/Type Ii 4 Phosphatase Rabbit Monoclonal Antibody
Cat # M06336
The listed rabbit monoclonal anti-INPP4B antibody, M06336, has reported human, mouse, and rat reactivity. Its Western blot image shows an approximately 105 kDa band in human MCF-7 whole cell lysate; the supplied evidence does not show mouse or rat blots.
Which to pick: M06336 is the only listed option. Its documented Western blot uses human MCF-7 whole cell lysate and a 1:1000 primary dilution. For mouse or rat samples, reactivity is listed, but no corresponding Western blot image is supplied.
Source: BosterBio INPP4B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O15327.
- Human Protein Atlas. INPP4B tissue expression.
- PMC8767114 — target-verified WB comparison
- PMC4578723 — target-verified WB comparison
- PMC11536254 — target-verified WB comparison