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- Table of Contents
RAB22A / Ras-related protein Rab-22A · Western blot design guide
Design a Western Blot for RAB22A
Source-linked RAB22A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RAB22A 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
RAB22A 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 | ~21.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (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 | 1 isoform(s) |
Section 1
Source-Linked RAB22A Western Blot Protocol Options
The M05967 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | (1) MCF-7 cell lysate; (2) BxPC-3 cell lysate (catalog M05967) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M05967; 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 RAB22A Western Blot Band Size?
RAB22A is predicted at 21.9 kDa; prenylation and membrane association are annotated, but their effects on migration have not been demonstrated here.
| Band near 21.9 kDa | consistent with the predicted RAB22A mass; confirm identity with controls |
| Band in a membrane fraction | consistent with membrane-associated RAB22A |
| Weak band in a soluble fraction | consistent with RAB22A partitioning with membranes |
| One principal band near 21.9 kDa | consistent with the single annotated isoform; identity still requires controls |
💡Expected RAB22A appearanceUniProt predicts RAB22A at 21.9 kDa, but no empirical band size is supplied; use an identity control to assess a band near that size.
| Predicted molecular mass | 21.9 kDa is the sequence-based reference size |
| Prenylation annotation | may affect migration, but no size shift is established |
| Lipid anchor | supports membrane association; no apparent size shift is established |
| Single annotated isoform | provides no basis for predicting an isoform size pattern |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | membrane-associated RAB22A may be poorly extracted | check membrane extraction and a positive-control lysate |
| Band higher than expected | the supplied features do not establish its identity or the size difference | compare with a RAB22A knockdown or other identity control |
| Band lower than expected | the supplied features do not establish cleavage or the band’s identity | check antibody specificity with a RAB22A knockdown |
| Multiple bands | the single annotated isoform does not explain additional bands | identify the RAB22A-dependent band with a knockdown control |
| Weak or no signal | membrane-associated RAB22A may be underrepresented after extraction | check recovery in a membrane fraction and use a positive control |
Experiment evidence
Western blot images showing RAB22A band
M05967 · RAB22A
M05967 · RAB22A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RAB22A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RAB22A in Western blot, you can use adrenal gland tissue, which has high HPA expression.
Positive control: Adrenal gland (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: Adipose tissue offers a negative control, but membrane-associated RAB22A requires efficient extraction.
HPA tissue expression evidence for RAB22A
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Lung | alveolar cells | Low | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced RAB22A Western Blot Tips
Deeper troubleshooting and optimisation questions for RAB22A, answered from its protein features.
How should RAB22A 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 annotated isoforms explain multiple RAB22A bands?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. The provided features therefore do not support assigning multiple bands to annotated isoforms.
What modification information matters when interpreting RAB22A bands?
PTM · RAB22A is described as lipid anchored, and prenylation is a keyword. No modified residue or glycosylation site is listed. These features do not establish that a modification causes a detectable band shift.
Does this guide establish induction of RAB22A?
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 RAB22A Western blot?
Transfer · RAB22A is a small protein with a predicted mass of 21.9 kDa. Choose transfer conditions that retain proteins near this size, then check both the membrane and post-transfer gel to assess recovery.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M05967 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 can localization affect RAB22A quantitation?
Quantitation · RAB22A is listed at endosome and cell membranes and is lipid anchored. Keep sample preparation and the fraction being measured consistent across samples; a change in membrane recovery could change the measured signal.
Should RAB22A migrate at exactly 21.9 kDa?
Interpretation · 21.9 kDa is the predicted mass. No observed band position is supplied, and the listed lipid anchor and prenylation keyword do not establish a visible shift. Compare the band with a molecular weight marker without treating an exact match as required.
The supplied features list one isoform and no observed band position. They do not identify the cause of additional bands. Check whether bands reproduce across samples and whether their abundance follows RAB22A-specific controls before assigning them to RAB22A.
The supplied features list one isoform and no observed band position. They do not identify the cause of additional bands. Check whether bands reproduce across samples and whether their abundance follows RAB22A-specific controls before assigning them to RAB22A.
Boster reagents
RAB22A 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-RAB22A Rabbit Monoclonal Antibody
Cat # M05967
The catalog reports M05967, a rabbit monoclonal anti-RAB22A antibody with reported Human, Mouse, and Rat reactivity. Its Western blot image shows RAB22A expression in MCF-7 and BxPC-3 cell lysates; the supplied evidence does not establish performance in other specimens.
Which to pick: M05967 is the only listed option. Its WB image uses MCF-7 and BxPC-3 lysates; check whether the reported reactivity and tested samples fit your experiment.
Source: BosterBio RAB22A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UL26.
- Human Protein Atlas. RAB22A tissue expression.
- PMC5342118 — target-verified WB comparison
- PMC11270584 — target-verified WB comparison
- PMC9217539 — target-verified WB comparison