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- Table of Contents
CRBN / Protein cereblon · Western blot design guide
Design a Western Blot for CRBN
Real validated CRBN Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CRBN 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
CRBN 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 | ~50.5 kDa | |
| Observed band | ~51 kDa | |
| Gel | 5–20% (catalog A00983-1) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated CRBN Western Blot Protocols
The A00983-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEL, human K562, human 293T (catalog A00983-1) |
| Gel % | 5–20% (catalog A00983-1) |
| Load | 30 ug; reducing conditions (catalog A00983-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00983-1) |
| Membrane | nitrocellulose membrane (catalog A00983-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00983-1) |
| Primary antibody | A00983-1 · 0.5 μg/mL (catalog A00983-1) |
| Primary incubation | overnight at 4°C (catalog A00983-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00983-1) |
| Secondary incubation | 1.5 hour at RT (catalog A00983-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00983-1) |
| Detection | ECL (catalog A00983-1) |
Section 2
What Is the Expected CRBN Western Blot Band Size?
CRBN is predicted at 50.5 kDa and observed at ~51 kDa; the cause of the small difference is not established.
| Sharp band at ~51 kDa | Matches the empirical CRBN band near its predicted mass. |
| Additional band above ~51 kDa | Could represent a CRBN isoform, but its identity requires validation. |
| Additional band below ~51 kDa | Could represent a CRBN isoform, but its identity requires validation. |
| Doublet near ~51 kDa | Could reflect isoforms 1 and 2; distinct migration has not been established. |
💡Expected CRBN appearanceCRBN has a predicted mass of 50.5 kDa and an empirical band at ~51 kDa; confirm band identity with ordinary controls, since migration of individual isoforms is not established.
| Predicted CRBN mass of 50.5 kDa | Places the full-length protein near the empirical ~51 kDa band. |
| Isoform 1 | Its individual mass and migration are not supplied. |
| Isoform 2 | Its individual mass and migration are not supplied. |
| Alternative splicing into isoforms 1 and 2 | Could yield different apparent sizes, but distinct bands are not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CRBN may be underrepresented in the sampled cellular fraction. | Check cytoplasmic, nuclear, and membrane fractions with appropriate controls. |
| Band higher than expected | An alternate isoform or unrelated antibody signal is possible. | Check band identity with CRBN depletion and a second antibody. |
| Band lower than expected | An alternate isoform or unrelated antibody signal is possible. | Check band identity with CRBN depletion and a second antibody. |
| Multiple bands | CRBN has two named isoforms, though their migration is unreported. | Test which bands decrease after CRBN depletion. |
| Weak or no signal | The sampled fraction may contain little CRBN. | Check sample loading and compare cellular fractions. |
Experiment evidence
Western blot images showing CRBN band
A00983-1 · CRBN
A00983 · CRBN
A00983 · CRBN
A00983 · CRBN
A00983 · CRBN
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CRBN Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CRBN in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for CRBN
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 | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced CRBN Western Blot Tips
Deeper troubleshooting and optimisation questions for CRBN, answered from its protein features.
How should CRBN 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 CRBN isoforms produce separate bands?
Isoforms · UniProt lists isoforms 1 and 2; isoform 2 lacks residue 23 of the canonical sequence. That small difference does not establish that the isoforms will resolve as separate bands. Check whether the antibody recognizes both isoforms before interpreting band intensity.
Where is the reported CRBN phosphorylation site?
PTM · UniProt lists phosphoserine at position 25 in the canonical sequence. Keep that numbering convention explicit when comparing antibody or paper annotations, which may use different coordinates.
Phosphoserine 25 is listed, but its presence alone does not demonstrate a visible shift. If a shifted band appears, compare matched samples with and without phosphatase treatment before assigning it to phosphorylation.
Phosphoserine 25 is listed, but its presence alone does not demonstrate a visible shift. If a shifted band appears, compare matched samples with and without phosphatase treatment before assigning it to phosphorylation.
Does this guide establish induction of CRBN?
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 CRBN?
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 A00983-1 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 CRBN bands be quantified across samples?
Quantitation · Quantify the band near the supplied observed position of about 51 kDa using consistent sample preparation and fraction selection. Because isoform 2 differs by one missing canonical residue, a single band may represent both isoforms; do not assign isoform-specific abundance from that band alone.
Why does CRBN appear near 51 kDa?
Interpretation · Its predicted mass is 50.5 kDa, close to the supplied observed band of about 51 kDa. The listed features do not establish a cause for any small difference between calculated and apparent mass.
CRBN is reported in the cytoplasm and nucleus and as a peripheral membrane protein. Consider these locations when selecting fractions; a weak band in one fraction alone does not establish low total CRBN abundance.
CRBN is a component of a ligase complex with CUL4A, DDB1 and RBX1, but complex membership alone does not identify a higher-mass Western-blot band. The supplied features also include a Ubl conjugation keyword without a listed conjugation site. Verify band identity before assigning either explanation.
CRBN is reported in the cytoplasm and nucleus and as a peripheral membrane protein. Consider these locations when selecting fractions; a weak band in one fraction alone does not establish low total CRBN abundance.
CRBN is a component of a ligase complex with CUL4A, DDB1 and RBX1, but complex membership alone does not identify a higher-mass Western-blot band. The supplied features also include a Ubl conjugation keyword without a listed conjugation site. Verify band identity before assigning either explanation.
Boster reagents
CRBN 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-CRBN Antibody Picoband®
Cat # A00983-1
Real WB data
Anti-Protein cereblon CRBN Antibody
Cat # A00983
Two the supplier anti-CRBN antibodies, A00983-1 and A00983, list human, mouse, and rat reactivity and have WB images. A00983-1 shows an approximately 51 kDa band in the listed cell and tissue lysates; A00983 shows human testis WB. No publication evidence is supplied.
Which to pick: For human HEL, K562, or 293T cells, or rat or mouse brain or heart, consider A00983-1 based on its WB image. For human testis, consider A00983, whose image uses 15 µg lysate. These examples do not establish performance in every sample.
Source: BosterBio CRBN gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96SW2.
- Human Protein Atlas. CRBN tissue expression.
- PMC6475089 — target-verified WB comparison
- PMC8228114 — target-verified WB comparison
- PMC13033450 — target-verified WB comparison