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- Table of Contents
RAMP2 / Receptor activity-modifying protein 2 · Western blot design guide
Design a Western Blot for RAMP2
Source-linked RAMP2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RAMP2 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
RAMP2 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 | ~19.6 kDa | |
| Gel | 15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked RAMP2 Western Blot Protocol Options
The A04768 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A04768) |
| Gel % | 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 | 0.2 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A04768) |
| Primary antibody | A04768 · 1:1000 (catalog A04768) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A04768) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04768) |
Section 2
What Is the Expected RAMP2 Western Blot Band Size?
RAMP2 has a predicted precursor mass of 19.6 kDa; signal-peptide cleavage, Asn130 glycosylation, and isoforms could affect migration, but no empirical band is supplied.
| Band near 19.6 kDa | consistent with the predicted RAMP2 precursor mass; confirm identity with antibody controls |
| Band below 19.6 kDa | may reflect removal of the signal peptide at residues 1–42 |
| Band above 19.6 kDa | N-linked glycosylation at Asn130 could affect migration, but a shift is not established |
| Multiple bands | isoforms 1 and 2 could contribute, but distinct migration is not established |
💡Expected RAMP2 appearanceThe predicted RAMP2 precursor mass is 19.6 kDa; signal-peptide removal, Asn130 glycosylation, and isoforms could affect migration, but no empirical band size is supplied, so verify identity with ordinary controls.
| UniProt predicted mass | places the full-length precursor at 19.6 kDa |
| Signal peptide at residues 1–42 | cleavage could make mature RAMP2 smaller than the precursor |
| N-linked glycosylation at Asn130 | could increase apparent size; a visible shift is not established |
| Isoforms 1 and 2 | could differ in size; their masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | membrane RAMP2 may be poorly recovered or solubilized | check membrane extraction and use a positive-control lysate |
| Band higher than expected | Asn130 glycosylation could affect migration | compare treated and untreated samples and confirm band identity |
| Band lower than expected | signal-peptide cleavage could reduce size | check antibody epitope coverage and confirm band identity |
| Multiple bands | isoforms 1 and 2 are listed, but separate bands are unproven | compare isoform-specific expression or depletion controls |
| Weak or no signal | membrane protein recovery may be low | verify extraction, loading, and antibody performance with a positive control |
Experiment evidence
Western blot images showing RAMP2 band
A04768 · RAMP2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RAMP2 Western blot
🧪For positive controls for RAMP2 in Western blot, you can use a verified RAMP2-positive sample; the supplied HPA data identify no tissue or cell line.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: RAMP2 is a membrane protein, and the missing HPA data prevent tissue-based control selection.
HPA tissue expression evidence for RAMP2
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced RAMP2 Western Blot Tips
Deeper troubleshooting and optimisation questions for RAMP2, answered from its protein features.
How should RAMP2 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 RAMP2 isoforms produce different bands?
Isoforms · Two isoforms are listed. At UniProt position 54, isoform 2 replaces E with EASVPT, adding five residues relative to isoform 1. The sequence difference may affect size, but the features do not show whether the isoforms resolve as separate bands.
Which glycosylation site matters for band assignment?
PTM · The supplied N-linked glycosylation site is Asn130 in UniProt numbering. Keep that coordinate convention when comparing papers or antibody information. A glycosylation annotation identifies a possible source of band variation; it does not establish the size or presence of a shifted band.
Does this guide establish induction of RAMP2?
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 RAMP2 Western blot?
Transfer · RAMP2 is a predicted 19.6-kDa, single-pass membrane protein. Choose transfer conditions that retain small proteins, and check transfer recovery when optimizing the blot. The supplied features do not specify a transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04768 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 RAMP2 bands be quantified across samples?
Quantitation · Quantify consistently assigned bands using matched sample preparation and detection conditions. Consider the two isoforms and the annotated Asn130 N-linked glycosylation site before combining bands. No empirical RAMP2 band position is supplied, so establish band identity before comparing amounts.
Why might a RAMP2 band differ from 19.6 kDa?
Interpretation · 19.6 kDa is the predicted mass for the 175-residue sequence. RAMP2 has a signal peptide at residues 1–42 and an N-linked glycosylation site at UniProt Asn130. Processing or glycosylation could affect apparent size, but these features alone do not establish a visible shift. No observed band size is supplied.
The annotated signal peptide spans UniProt residues 1–42. Account for possible signal-peptide removal when comparing a band with the predicted mass of the full sequence. The supplied features do not give a measured mass for processed RAMP2.
RAMP2 has two annotated disulfide bonds. Compare reducing and nonreducing samples under otherwise matched conditions if band behavior differs. The annotation alone does not predict a specific mobility change.
RAMP2 is annotated as a glycoprotein and forms heterodimers with CALCRL or CALCR. These features suggest possibilities to investigate, but a higher band alone cannot identify a glycosylated form or receptor complex. Compare reproducible bands across conditions and verify their RAMP2 identity.
The annotated signal peptide spans UniProt residues 1–42. Account for possible signal-peptide removal when comparing a band with the predicted mass of the full sequence. The supplied features do not give a measured mass for processed RAMP2.
RAMP2 has two annotated disulfide bonds. Compare reducing and nonreducing samples under otherwise matched conditions if band behavior differs. The annotation alone does not predict a specific mobility change.
RAMP2 is annotated as a glycoprotein and forms heterodimers with CALCRL or CALCR. These features suggest possibilities to investigate, but a higher band alone cannot identify a glycosylated form or receptor complex. Compare reproducible bands across conditions and verify their RAMP2 identity.
Boster reagents
RAMP2 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-RAMP2 Antibody
Cat # A04768
The catalog reports A04768, an anti-RAMP2 antibody with reported Human, Mouse, and Rat reactivity. Its Western blot image uses extracts from various cell lines at 1:1000 dilution and 25 µg protein per lane; the caption does not identify the cell lines.
Which to pick: A04768 is the only listed option and has a Western blot image. Check its reported reactivity against your sample species; the image caption does not establish which species were tested.
Source: BosterBio RAMP2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O60895.
- Human Protein Atlas. RAMP2 tissue expression.
- PMC4022580 — target-verified WB comparison
- PMC7760190 — target-verified WB comparison