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- Table of Contents
RARA / Retinoic acid receptor alpha · Western blot design guide
Design a Western Blot for RARA
Real validated RARA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RARA 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
RARA 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 | ~50.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| 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 Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated RARA Western Blot Protocols
The M00392 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MCF-7 cell lysate (catalog M00392) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M00392; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected RARA Western Blot Band Size?
RARA is predicted at 50.8 kDa; isoforms and phosphorylation may affect migration, but no distinct band positions are demonstrated by the supplied evidence.
| Band near 50.8 kDa | consistent with the predicted RARA mass; identity requires confirmation |
| Several bands at different positions | could reflect Alpha-1, Alpha-2 and Alpha-1-deltaBC if their migration differs and identities are confirmed |
| Band in a nuclear fraction | consistent with RARA nuclear localization |
| Band in a cytoplasmic fraction | consistent with RARA cytoplasmic localization |
💡Expected RARA appearanceRARA has a predicted mass of 50.8 kDa, but no empirical band size is supplied; confirm any candidate band with an independent antibody or other identity control.
| Predicted RARA mass | 50.8 kDa provides the reference size, not a measured band position |
| Alpha-1 splice isoform | isoform-specific mass and migration are not supplied |
| Alpha-2 splice isoform | size may differ from other isoforms, but its mass and migration are not supplied |
| Alpha-1-deltaBC splice isoform | size may differ from other isoforms, but its mass and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RARA may be weak in the sampled fraction | check matched nuclear and cytoplasmic fractions with a validated antibody |
| Band higher than expected | the cause of altered migration is unestablished; RARA has documented phosphorylation sites | compare phosphatase-treated material and confirm identity with an independent antibody |
| Band lower than expected | a splice isoform is possible, but isoform-specific sizes are unknown | check isoform expression and confirm the band with an independent antibody |
| Multiple bands | Alpha-1, Alpha-2 and Alpha-1-deltaBC are annotated isoforms, but distinct migration is unproven | verify isoform expression and band identity independently |
| Weak or no signal | RARA distribution between nucleus and cytoplasm can vary with ligand binding and modifications | compare matched nuclear and cytoplasmic fractions and check antibody performance |
Experiment evidence
Western blot images showing RARA band
M00392 · RARA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RARA Western blot
🧪For positive controls for RARA in Western blot, you can use a verified RARA-expressing sample; the supplied HPA evidence identifies no positive tissue or cell.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no tissue data, so a tissue-based positive or negative control cannot be selected from this evidence.
HPA tissue expression evidence for RARA
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced RARA Western Blot Tips
Deeper troubleshooting and optimisation questions for RARA, answered from its protein features.
How should RARA 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.
Which RARA isoforms could affect band interpretation?
Isoforms · UniProt lists Alpha-1, Alpha-2 and Alpha-1-deltaBC. Alpha-2 replaces residues 1–60, while Alpha-1-deltaBC lacks residues 61–157. Consider these sequence differences when assessing bands, but do not assign a band to an isoform by size alone.
An antibody targeting residues 61–157 of canonical RARA would not recognize that segment in Alpha-1-deltaBC. The first 60 residues also differ in Alpha-2. Check the antibody's stated epitope against each isoform sequence before comparing signals.
An antibody targeting residues 61–157 of canonical RARA would not recognize that segment in Alpha-1-deltaBC. The first 60 residues also differ in Alpha-2. Check the antibody's stated epitope against each isoform sequence before comparing signals.
Which RARA phosphorylation sites matter when interpreting bands?
PTM · UniProt annotates phosphoserine at positions 77 by CDK7, 96 by PKB/AKT1, and 219 and 369 by PKA. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. Alpha-1-deltaBC lacks canonical residues 61–157, including positions 77 and 96. These annotations do not establish a visible band shift.
Does this guide establish induction of RARA?
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 RARA?
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 M00392 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 RARA 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.
Should RARA run exactly at its predicted 50.8 kDa?
Interpretation · 50.8 kDa is the predicted mass; no observed band position is supplied. Phosphorylation and alternative isoforms are annotated, but their presence alone does not establish a visible shift or explain a difference from 50.8 kDa.
The supplied feature says pulsatile shear stress enhances RARA association with RXRA through its C-terminus. It does not establish increased RARA abundance. Measure RARA signal under matched conditions if abundance is the question.
RARA is annotated in both nucleus and cytoplasm. Its nuclear localization depends on ligand binding, phosphorylation and sumoylation. Compare like fractions under matched conditions; a changed nuclear signal alone does not establish a change in total RARA abundance.
Check whether the antibody epitope is retained in the three annotated isoforms, especially the 61–157 deletion in Alpha-1-deltaBC. Consider the listed phosphorylation sites, but do not assign an unexpected band to phosphorylation or an isoform from apparent size alone; no empirical band position is supplied.
The supplied feature says pulsatile shear stress enhances RARA association with RXRA through its C-terminus. It does not establish increased RARA abundance. Measure RARA signal under matched conditions if abundance is the question.
RARA is annotated in both nucleus and cytoplasm. Its nuclear localization depends on ligand binding, phosphorylation and sumoylation. Compare like fractions under matched conditions; a changed nuclear signal alone does not establish a change in total RARA abundance.
Check whether the antibody epitope is retained in the three annotated isoforms, especially the 61–157 deletion in Alpha-1-deltaBC. Consider the listed phosphorylation sites, but do not assign an unexpected band to phosphorylation or an isoform from apparent size alone; no empirical band position is supplied.
Boster reagents
RARA 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-Retinoic Acid Receptor alpha RARA Rabbit Monoclonal Antibody
Cat # M00392
M00392 is an anti-RARA rabbit monoclonal antibody listed for Western blot with human and mouse reactivity. Its supplied WB image shows RARA expression in MCF-7 cell lysate; no other tested samples are documented here.
Which to pick: M00392 is the only listed option. Its WB image uses MCF-7 cell lysate; consider your sample against the stated human and mouse reactivity, since the supplied image does not document other sample contexts.
Source: BosterBio RARA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P10276.
- Human Protein Atlas. RARA tissue expression.
- PMC6838077 — target-verified WB comparison
- PMC3759040 — target-verified WB comparison
- PMC6711153 — target-verified WB comparison