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HMGCR / 3-hydroxy-3-methylglutaryl-coenzyme A reductase · Western blot design guide
Design a Western Blot for HMGCR
Real validated HMGCR Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-HMGCR 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
HMGCR 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 | ~97.5 kDa | |
| Observed band | ~97 kDa | |
| Gel | 5–20% (catalog A00643-3) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated HMGCR Western Blot Protocols
The A00643-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human CACO-2, human RT4, human THP-1 (catalog A00643-3) |
| Gel % | 5–20% (catalog A00643-3) |
| Load | 30 ug; reducing conditions (catalog A00643-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00643-3) |
| Membrane | nitrocellulose membrane (catalog A00643-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00643-3) |
| Primary antibody | A00643-3 · 0.5 μg/mL (catalog A00643-3) |
| Primary incubation | overnight at 4°C (catalog A00643-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00643-3) |
| Secondary incubation | 1.5 hour at RT (catalog A00643-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00643-3) |
| Detection | ECL (catalog A00643-3) |
Section 2
What Is the Expected HMGCR Western Blot Band Size?
HMGCR is predicted at 97.5 kDa and observed at ~97 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~97 kDa | Empirical HMGCR band, close to the 97.5 kDa predicted mass |
| Band near twice the monomer size | Could reflect a homodimer retained during preparation; HMGCR has no listed disulfide bonds |
| Additional bands at different sizes | Could reflect isoforms 1, 2, or 3; their migration is not supplied |
| Shifted band near ~97 kDa | Could reflect N-linked modification at Asn281 or Asn296; a visible shift is not established |
💡Expected HMGCR appearanceHMGCR is predicted at 97.5 kDa and has an empirical band at ~97 kDa; confirm band identity with an HMGCR-specific control because the listed modifications and isoforms do not establish distinct migration patterns.
| Predicted HMGCR mass | 97.5 kDa for the supplied sequence |
| N-linked glycosylation at Asn281 | Could alter apparent size; no visible shift is established |
| N-linked glycosylation at Asn296 | Could alter apparent size; no visible shift is established |
| Homodimer formation | Could appear near twice the monomer size if retained during preparation |
| Isoforms 1, 2, and 3 | May differ in size; their relative masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | HMGCR is a multi-pass membrane protein that may be poorly recovered | Check membrane-protein extraction and a positive lysate control |
| Band higher than expected | A retained homodimer or altered migration is possible; the cause is unproven | Compare denaturing conditions and verify identity with an HMGCR-specific control |
| Band lower than expected | An isoform or sample degradation is possible; no fragment size is supplied | Compare antibody epitopes and check sample handling with a positive control |
| Multiple bands | Isoforms 1, 2, and 3 are listed, but distinct bands are not established | Confirm each band with an independent HMGCR antibody or knockdown control |
| Weak or no signal | Recovery of membrane-associated HMGCR may be low | Check extraction, loading, transfer, and a positive lysate control |
Experiment evidence
Western blot images showing HMGCR band
A00643-3 · HMGCR
M00643 · HMGCR
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for HMGCR Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for HMGCR in Western blot, you can use appendix lysate, which HPA scores High.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a membrane protein, HMGCR may give a stronger signal in membrane-enriched lysate.
HPA tissue expression evidence for HMGCR
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced HMGCR Western Blot Tips
Deeper troubleshooting and optimisation questions for HMGCR, answered from its protein features.
How should HMGCR 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 HMGCR isoforms produce different bands?
Isoforms · Three isoforms are listed. Isoform 2 lacks canonical residues 522–574, while isoform 3 has an altered N terminus. Check whether the antibody recognizes a region retained in the isoform you intend to measure. These sequence differences do not establish where either isoform will migrate.
Which glycosylation sites matter when interpreting bands?
PTM · UniProt lists N-linked sites at Asn281 and Asn296. These are canonical-sequence coordinates; other numbering conventions may differ. Their presence supports considering glycosylation when assessing an unexpected band, but does not demonstrate a visible shift.
UniProt lists phosphoserine at canonical Ser504 and Ser872. State that coordinate convention when comparing antibody or paper numbering. The listed sites do not by themselves predict separate Western blot bands.
UniProt lists phosphoserine at canonical Ser504 and Ser872. State that coordinate convention when comparing antibody or paper numbering. The listed sites do not by themselves predict separate Western blot bands.
Does this guide establish induction of HMGCR?
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 HMGCR Western blot?
Transfer · HMGCR is an 888-residue, multi-pass ER and peroxisome membrane protein. Verify recovery of the membrane-associated protein and transfer of the approximately 97 kDa band before interpreting a weak signal. The supplied features do not establish a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00643-3 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 HMGCR 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.
Why is HMGCR near 97 kDa?
Interpretation · The observed band near 97 kDa agrees with the supplied 97.5 kDa predicted mass. The listed modifications alone do not establish a visible shift or explain a mass difference.
The supplied interaction record says sterols accelerate HMGCR binding to INSIG1 and recruitment to AMFR/gp78. Record sterol treatment consistently across samples when comparing HMGCR bands. That interaction alone does not specify the size or direction of a band-intensity change.
Compare the same approximately 97 kDa band across samples prepared and transferred consistently, accounting for HMGCR's membrane localization. If an antibody could detect multiple isoforms, define which band is quantified and check that its target sequence is retained.
Consider the three listed isoforms, the Asn281 and Asn296 glycosylation sites, and phosphoserine at Ser504 and Ser872. Check antibody recognition against isoform 2's deletion of canonical residues 522–574. None of these features alone identifies an unexpected band.
The supplied interaction record says sterols accelerate HMGCR binding to INSIG1 and recruitment to AMFR/gp78. Record sterol treatment consistently across samples when comparing HMGCR bands. That interaction alone does not specify the size or direction of a band-intensity change.
Compare the same approximately 97 kDa band across samples prepared and transferred consistently, accounting for HMGCR's membrane localization. If an antibody could detect multiple isoforms, define which band is quantified and check that its target sequence is retained.
Consider the three listed isoforms, the Asn281 and Asn296 glycosylation sites, and phosphoserine at Ser504 and Ser872. Check antibody recognition against isoform 2's deletion of canonical residues 522–574. None of these features alone identifies an unexpected band.
Boster reagents
HMGCR 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-HMGCR Antibody Picoband®
Cat # A00643-3
Real WB data
Anti-HMGCR/Hmg Coa Reductase Rabbit Monoclonal Antibody
Cat # M00643
Both listed anti-HMGCR antibodies have WB images: A00643-3 in human CACO-2, RT4, and THP-1 lysates with an approximately 97 kDa band, and M00643 in Jurkat lysate. The supplied M00643 caption gives limited conditions and no mouse or rat WB example.
Which to pick: For human WB with reported loading, antibody concentration, and band size, consider A00643-3. M00643 lists human, mouse, and rat reactivity and has a Jurkat WB image, but the supplied caption does not document mouse or rat WB testing.
Source: BosterBio HMGCR gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P04035.
- Human Protein Atlas. HMGCR tissue expression.
- PMC4503878 — target-verified WB comparison
- PMC12615409 — target-verified WB comparison
- PMC13534982 — target-verified WB comparison