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- Table of Contents
RBP1 / Retinol-binding protein 1 · Western blot design guide
Design a Western Blot for RBP1
Real validated RBP1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RBP1 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
RBP1 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 | ~15.9 kDa | |
| Observed band | ~16 kDa | |
| Gel | 5–20% (catalog A03820-4) | |
| Positive control | Testis (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 | Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated RBP1 Western Blot Protocols
The A03820-4 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SH-SY5Y, rat liver (catalog A03820-4) |
| Gel % | 5–20% (catalog A03820-4) |
| Load | 30 ug; reducing conditions (catalog A03820-4) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03820-4) |
| Membrane | nitrocellulose membrane (catalog A03820-4) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03820-4) |
| Primary antibody | A03820-4 · 0.5 μg/mL (catalog A03820-4) |
| Primary incubation | overnight at 4°C (catalog A03820-4) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03820-4) |
| Secondary incubation | 1.5 hour at RT (catalog A03820-4) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03820-4) |
| Detection | ECL (catalog A03820-4) |
Section 2
What Is the Expected RBP1 Western Blot Band Size?
RBP1 is predicted at 15.9 kDa and observed at ~16 kDa; its three isoforms have no supplied migration data.
| Single band at ~16 kDa | Matches the observed RBP1 band and its 15.9 kDa predicted mass. |
| Additional bands at other positions | Could reflect isoforms 1, 2, or 3; their migration has not been established. |
| Band in a cytoplasmic fraction | Consistent with RBP1's cytoplasmic location. |
| Little or no band in an extracellular fraction | Consistent with RBP1's reported intracellular locations. |
💡Expected RBP1 appearanceRBP1 has a predicted mass of 15.9 kDa and an observed band at ~16 kDa; confirm band identity with antibody and sample controls if other bands appear.
| Predicted RBP1 mass | 15.9 kDa predicted; the observed band is ~16 kDa. |
| Isoform 1 | May differ in size from other isoforms; its relative size is unknown. |
| Isoform 2 | May differ in size from other isoforms; its relative size is unknown. |
| Isoform 3 | May differ in size from other isoforms; its relative size is unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | The preparation may lack cytoplasmic or lipid droplet RBP1. | Check fraction content and include a positive whole-cell lysate control. |
| Band higher than expected | Its identity is uncertain; an isoform size difference is possible but unverified. | Compare with a positive control and verify antibody specificity. |
| Band lower than expected | Its identity is uncertain; isoform migration is not documented. | Compare with a positive control and verify antibody specificity. |
| Multiple bands | RBP1 has three named isoforms, but distinct band positions are unverified. | Check isoform expression and validate each band with an independent antibody or RBP1 depletion. |
| Weak or no signal | RBP1 may be poorly recovered from the cytoplasmic or lipid droplet fraction. | Check extraction and loading against a positive lysate control. |
Experiment evidence
Western blot images showing RBP1 band
A03820-4 · RBP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RBP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RBP1 in Western blot, you can use testis lysate, which has high HPA expression.
Positive control: Testis (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Cytoplasmic RBP1 should be accessible in tissue lysate, with adipose tissue providing an HPA not-detected comparison.
HPA tissue expression evidence for RBP1
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 |
|---|---|---|---|---|
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Fallopian tube | ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Ovary | ovarian stroma 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | basal cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RBP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for RBP1, answered from its protein features.
How should RBP1 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 RBP1 isoforms produce different bands?
Isoforms · Three isoforms are listed. Isoforms 2 and 3 replace the initial methionine with a longer sequence and replace residues 85–135 with different short sequences. These differences could affect migration, but the features alone do not establish that separate bands will be visible.
Check the antibody epitope against all three isoform sequences. Isoforms 2 and 3 differ at the N terminus and replace canonical residues 85–135, so an antibody targeting either altered region may detect the isoforms differently.
Check the antibody epitope against all three isoform sequences. Isoforms 2 and 3 differ at the N terminus and replace canonical residues 85–135, so an antibody targeting either altered region may detect the isoforms differently.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of RBP1?
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 RBP1 Western blot?
Transfer · RBP1 is predicted at 15.9 kDa. Choose transfer conditions and a membrane validated to retain proteins near 16 kDa, and check transfer before interpreting a missing band. The supplied features do not identify a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03820-4 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 RBP1 be quantified across cell fractions?
Quantitation · RBP1 is annotated in the cytoplasm and on lipid droplets. Compare like fractions across samples and account for both locations when interpreting a change in the measured signal.
Why is the RBP1 band near 16 kDa?
Interpretation · Canonical RBP1 is 135 amino acids with a predicted mass of 15.9 kDa, consistent with the reported apparent band near 16 kDa. The supplied features do not establish a cause for any small difference between calculated and apparent mass.
Methylation appears as a UniProt keyword, but no modified residue or coordinate is listed. Do not assign a shifted band to methylation from these features alone.
RBP1 interacts with STRA6 only as retinol-free apoprotein, but that interaction does not establish a higher RBP1 band on a Western blot. The listed isoform sequence changes also do not establish a specific band position. Confirm an unexpected band's identity before assigning either explanation.
Methylation appears as a UniProt keyword, but no modified residue or coordinate is listed. Do not assign a shifted band to methylation from these features alone.
RBP1 interacts with STRA6 only as retinol-free apoprotein, but that interaction does not establish a higher RBP1 band on a Western blot. The listed isoform sequence changes also do not establish a specific band position. Confirm an unexpected band's identity before assigning either explanation.
Boster reagents
RBP1 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-RBP1 Antibody Picoband®
Cat # A03820-4
The catalog reports one anti-RBP1 antibody, A03820-4, with stated human, mouse, and rat reactivity. Its WB image shows an approximately 16 kDa band in human SH-SY5Y cell and rat liver lysates; no mouse WB example is supplied.
Which to pick: A03820-4 is the only listed option. Its WB image uses human SH-SY5Y and rat liver lysates at 0.5 μg/mL primary antibody. Mouse reactivity is listed, but the supplied image does not test mouse samples.
Source: BosterBio RBP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P09455.
- Human Protein Atlas. RBP1 tissue expression.
- PMC12481618 — target-verified WB comparison
- PMC7316825 — target-verified WB comparison