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- Table of Contents
CREM / cAMP-responsive element modulator · Western blot design guide
Design a Western Blot for CREM
Real validated CREM Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CREM 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
CREM 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 | ~37 kDa | |
| Gel | 12–15% (standard starting point) | |
| 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 | Phosphorylated + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 29 isoform(s) |
Section 1
Real Curated CREM Western Blot Protocols
The A04379-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | lysates from RAW264.7 cells, (catalog A04379-1) |
| 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 | the synthesized peptide (catalog A04379-1) |
| Primary antibody | A04379-1; 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 CREM Western Blot Band Size?
CREM has a predicted mass of 37 kDa; isoforms and phosphorylation could affect its pattern, but their effects on migration are not demonstrated here.
| Band near 37 kDa | consistent with the predicted CREM mass; confirm identity with a blocking control |
| Several bands at different positions | may reflect CREM isoforms 1 through 29; their migration differences are unknown |
| Closely spaced doublet | could reflect CREM phosphorylation, but a visible shift is not established |
| No band in whole-cell lysate | nuclear CREM may be poorly recovered; check a nuclear extract |
💡Expected CREM appearanceUniProt predicts 37 kDa for CREM; no empirical band size is supplied, and isoform or phosphorylation effects on migration remain unverified, so confirm any candidate band with a blocking or depletion control.
| UniProt predicted mass | places the reference CREM sequence near 37 kDa |
| Splice isoforms 1 through 10 | may differ in size from the reference sequence; relative masses are not supplied |
| Splice isoforms 11 through 20 | may differ in size from other CREM isoforms; relative masses are not supplied |
| Splice isoforms 21 through 29 | may differ in size from other CREM isoforms; relative masses are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear CREM may be poorly recovered | check a nuclear extract and verify extraction with a nuclear marker |
| Band higher than expected | isoform identity or migration is unresolved | compare with a CREM depletion control; do not infer a dimer from the DNA-binding annotation |
| Band lower than expected | a different CREM isoform is possible, but its mass is unknown | confirm identity by CREM depletion or peptide blocking |
| Multiple bands | CREM has 29 annotated isoforms; phosphorylation is also annotated | use depletion or peptide blocking to identify CREM bands before assigning isoforms or phosphorylation |
| Weak or no signal | nuclear CREM may be underrepresented in the sample | check nuclear extraction and include a nuclear marker |
Experiment evidence
Western blot images showing CREM band
A04379-1 · CREM
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CREM Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CREM 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.
⚠️Feasibility: A tissue negative is feasible because HPA lists adipose tissue as not detected.
HPA tissue expression evidence for CREM
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 | round or early spermatids | High | Protein (IHC) | HPA → |
| Appendix | goblet cells | Medium | Protein (IHC) | HPA → |
| Colon | goblet cells | Medium | Protein (IHC) | HPA → |
| Duodenum | goblet cells | Medium | Protein (IHC) | HPA → |
| Endometrium | 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CREM Western Blot Tips
Deeper troubleshooting and optimisation questions for CREM, answered from its protein features.
How should CREM 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.
How could CREM isoforms affect the band pattern?
Isoforms · UniProt lists 29 isoforms with substantial sequence differences. For example, isoform 20 lacks canonical residues 1–242, while isoform 22 lacks residues 187–345. Different isoforms may therefore produce distinct bands; the feature list does not establish which isoforms are present in your sample.
Which CREM phosphorylation sites matter for phospho-specific detection?
PTM · The listed phosphoserines are at UniProt canonical positions 102, 129, 271, 274 and 277. Check which of these residues each target isoform retains before selecting a phospho-specific antibody. Paper or antibody numbering may differ from these UniProt coordinates.
Does this guide establish induction of CREM?
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 CREM?
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 A04379-1 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 multiple CREM bands be quantified?
Quantitation · Define the band or isoforms being measured and confirm that the antibody recognizes their retained sequences. Quantify distinct bands separately unless their identities and antibody detection are established. A combined signal could mix products from the 29 listed isoforms.
Should CREM run exactly at its predicted 37 kDa?
Interpretation · 37 kDa is the supplied predicted mass; no observed band position is available. Compare candidate bands with this estimate, but do not assign identity from migration alone. The listed splice variants and phosphorylation sites do not establish a visible shift or explain a particular apparent mass.
Match the antibody epitope to the isoforms you intend to detect. Isoforms 21 and 24 replace canonical residues 1–244, and isoform 22 lacks residues 187–345. An antibody against a missing segment could overlook those isoforms.
Consider annotated sequence variants: isoform 25 lacks canonical residues 1–195, and isoform 16 lacks residues 42–232. Check whether the antibody epitope survives in the candidate isoform. These annotations suggest possibilities but do not identify an observed band.
UniProt lists CREM in the nucleus. A nuclear fraction is therefore relevant when assessing nuclear CREM; document the fraction used when comparing results. The supplied features do not establish that every isoform is confined to that fraction.
Match the antibody epitope to the isoforms you intend to detect. Isoforms 21 and 24 replace canonical residues 1–244, and isoform 22 lacks residues 187–345. An antibody against a missing segment could overlook those isoforms.
Consider annotated sequence variants: isoform 25 lacks canonical residues 1–195, and isoform 16 lacks residues 42–232. Check whether the antibody epitope survives in the candidate isoform. These annotations suggest possibilities but do not identify an observed band.
UniProt lists CREM in the nucleus. A nuclear fraction is therefore relevant when assessing nuclear CREM; document the fraction used when comparing results. The supplied features do not establish that every isoform is confined to that fraction.
Boster reagents
CREM 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-CREM Antibody
Cat # A04379-1
The catalog reports one anti-CREM antibody, A04379-1, with reported Human, Mouse, and Rat reactivity. Its Western blot image shows RAW264.7 cell lysates and a right lane blocked with synthesized peptide. The supplied evidence covers this example, not broad sample validation.
Which to pick: A04379-1 is the only listed option. Check its reported reactivity against your sample species; the available Western blot image documents RAW264.7 lysates with a peptide-blocked lane.
Source: BosterBio CREM gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q03060.
- Human Protein Atlas. CREM tissue expression.
- PMC4011562 — target-verified WB comparison
- PMC7044032 — target-verified WB comparison
- PMC7343452 — target-verified WB comparison