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- Table of Contents
CALCR / Calcitonin receptor · Western blot design guide
Design a Western Blot for CALCR
Source-linked CALCR Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CALCR 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
CALCR 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 | ~55.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Hypothalamus (IHC candidate; verify WB) +2 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 6 isoform(s) |
Section 1
Source-Linked CALCR Western Blot Protocol Options
The A03322-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | lysate from MCF-7, A431 cell line, (catalog A03322-1) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03322-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | 1:5000 (catalog A03322-1) |
| 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 CALCR Western Blot Band Size?
CALCR has a predicted 55.3 kDa precursor; glycosylation, signal-peptide cleavage and isoforms may affect migration, but no empirical band size is supplied.
| Band near 55.3 kDa | consistent with the predicted full-length precursor; confirm identity with controls |
| Band above 55.3 kDa | may reflect N-linked glycosylation at Asn28, Asn73, Asn125 or Asn130 |
| Band below 55.3 kDa | may reflect removal of the 1–24 signal peptide |
| Multiple bands | could reflect isoforms 1–6, though distinct migration is unestablished |
| Weak or absent lysate band | the membrane receptor may be poorly recovered during extraction |
💡Expected CALCR appearanceUniProt predicts a 55.3 kDa full-length CALCR precursor; signal-peptide cleavage, N-linked glycosylation and isoforms may alter migration, but no empirical band size is supplied, so confirm band identity with controls.
| Predicted full-length mass | sets a 55.3 kDa precursor reference, not a measured band |
| N-linked glycosylation at Asn28 | may increase apparent size if occupied |
| N-linked glycosylation at Asn73 | may increase apparent size if occupied |
| N-linked glycosylation at Asn125 | may increase apparent size if occupied |
| N-linked glycosylation at Asn130 | may increase apparent size if occupied |
| Signal peptide at residues 1–24 | cleavage yields a smaller mature chain than the precursor |
| Isoforms 1, 2, 3, 4, 5 and 6 | may differ in size; their masses and separation are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CALCR is a multi-pass membrane protein that may be poorly extracted | Check membrane protein recovery and use a membrane-enriched preparation |
| Band higher than expected | N-linked glycans at the four listed asparagines may affect migration | Compare matched samples before and after N-glycan removal |
| Band lower than expected | Signal-peptide cleavage can reduce size relative to the full-length precursor | Check the antibody epitope and verify band identity by peptide competition |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unconfirmed | Compare matched samples before and after N-glycan removal |
| Multiple bands | CALCR has six named isoforms, but their migration is unspecified | Verify candidate bands by peptide competition and compare isoform expression |
| Weak or no signal | Recovery of the multi-pass membrane receptor may be inefficient | Assess extraction and membrane enrichment before interpreting absence |
Experiment evidence
Western blot images showing CALCR band
A03322-1 · CALCR
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CALCR Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CALCR in Western blot, you can use hypothalamus tissue, which has high HPA expression.
Positive control: Hypothalamus (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 the samples.
⚠️Feasibility: As a multi-pass membrane protein, CALCR may require membrane-enriched lysate for a clear signal.
HPA tissue expression evidence for CALCR
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 |
|---|---|---|---|---|
| Hypothalamus | neuronal cells | High | Protein (IHC) | HPA → |
| Kidney | distal tubules | Medium | Protein (IHC) | HPA → |
| Placenta | syncytiotrophoblasts - cell body | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CALCR Western Blot Tips
Deeper troubleshooting and optimisation questions for CALCR, answered from its protein features.
How should CALCR 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 CALCR isoforms could produce different bands?
Isoforms · UniProt lists six isoforms. Isoforms 3 and 4 lack residues 1–47; isoforms 2, 4 and 6 replace residue 174 with RKLTTIFPLNWKYRKAL. Isoforms 5 and 6 replace 268–274 and lack 275–474. Use the expected isoform in your sample and the antibody epitope when assigning bands. Coordinates refer to the supplied UniProt sequence.
How should CALCR glycosylation affect band interpretation?
PTM · CALCR has four annotated N-linked sites: Asn28, Asn73, Asn125 and Asn130 in UniProt coordinates. Consider these sites when investigating multiple bands, but do not assign a band to glycosylation from migration alone. Isoforms 3 and 4 lack residues 1–47, which includes Asn28.
Does this guide establish induction of CALCR?
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 CALCR Western blot?
Transfer · CALCR is annotated as a multi-pass cell-membrane protein. The supplied features do not specify a transfer method. Choose transfer conditions suitable for your gel and apparatus, then check whether protein remains in the gel and whether the candidate CALCR band reached the membrane.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03322-1 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 CALCR 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 might CALCR migrate differently from its predicted 55.3 kDa?
Interpretation · The 55.3 kDa prediction does not account for every processed or modified form. CALCR has an annotated signal peptide at residues 1–24 and N-linked glycosylation sites at 28, 73, 125 and 130. These features warrant checking band identity, but their presence alone does not establish a visible shift or explain a specific apparent mass.
Quantify the same identified CALCR band across samples. The six annotated isoforms include major sequence differences, so check whether the antibody epitope is present in each isoform being compared. Keep the band definition consistent across lanes.
First compare each band's apparent size with the six annotated isoforms and the 55.3 kDa prediction. Check whether the antibody epitope falls within a region absent or changed in an isoform. The signal peptide and four N-linked sites provide additional possibilities to investigate; the supplied features alone cannot identify an unexpected band.
Quantify the same identified CALCR band across samples. The six annotated isoforms include major sequence differences, so check whether the antibody epitope is present in each isoform being compared. Keep the band definition consistent across lanes.
First compare each band's apparent size with the six annotated isoforms and the 55.3 kDa prediction. Check whether the antibody epitope falls within a region absent or changed in an isoform. The signal peptide and four N-linked sites provide additional possibilities to investigate; the supplied features alone cannot identify an unexpected band.
Boster reagents
CALCR 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-CALCR Antibody (C-term)
Cat # A03322-1
A03322-1 is a human-reactive anti-CALCR antibody with a Western blot image using MCF-7 and A431 lysates, with and without peptide. The supplied evidence is limited to this example.
Which to pick: A03322-1 is the only listed option. Its WB example uses MCF-7 and A431 lysates at 35 µg per lane and a 1:1000 primary antibody dilution; assess its suitability for your sample.
Source: BosterBio CALCR gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P30988.
- Human Protein Atlas. CALCR tissue expression.
- PMC11345353 — target-verified WB comparison
- PMC5280875 — target-verified WB comparison
- PMC7576311 — target-verified WB comparison