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- Table of Contents
CD177 / CD177 antigen · Western blot design guide
Design a Western Blot for CD177
Source-linked CD177 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CD177 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
CD177 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 | ~46.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | Glycosylated + Cleaved | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked CD177 Western Blot Protocol Options
The A03958-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Mouse bone marrow, (catalog A03958-1) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A03958-1) |
| Primary antibody | A03958-1 · 1:840 (catalog A03958-1) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A03958-1) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A03958-1) |
Section 2
What Is the Expected CD177 Western Blot Band Size?
CD177 has a predicted 46.4 kDa precursor; processing, Asn189 glycosylation and isoforms may affect migration, but no band position is demonstrated.
| Band near 46.4 kDa | consistent with the predicted precursor mass, pending identity controls |
| Band below 46.4 kDa | could reflect signal peptide or propeptide removal |
| Bands at different positions | could reflect isoforms 1, 2 and 3; distinct migration is unconfirmed |
| Little or no band in soluble lysate | consistent with membrane localization of CD177 |
💡Expected CD177 appearanceUniProt predicts a 46.4 kDa precursor; processing, N-linked glycosylation at Asn189 and isoforms may affect migration, but no empirical band size is supplied, so confirm any candidate band with identity controls.
| UniProt predicted mass | 46.4 kDa for the precursor, not a measured band position |
| N-linked glycosylation at Asn189 | may alter apparent mass; the size effect is not established |
| Isoforms 1, 2 and 3 | may differ in size; their masses and migration are not supplied |
| Signal peptide at residues 1–21 | cleavage yields a smaller mature product than the precursor |
| Propeptide at residues 409–437 | removal yields a smaller mature product than the precursor |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CD177 is membrane-associated and may be scarce in a soluble fraction | check a membrane-enriched fraction and a positive control |
| Band higher than expected | N-linked glycosylation at Asn189 may affect migration, but its size effect is unknown | compare glycosidase-treated and untreated samples with an identity control |
| Band lower than expected | signal peptide or propeptide cleavage could reduce size | check epitope placement and verify band identity |
| Multiple bands | isoforms 1, 2 and 3 are annotated, but separate bands are not established | compare isoform-aware controls and check antibody specificity |
| Weak or no signal | membrane localization may limit recovery in the tested fraction | enrich the membrane fraction and confirm detection with a positive control |
Experiment evidence
Western blot images showing CD177 band
A03958-1 · CD177
A03958-1 · CD177
A03958 · CD177
A03958 · CD177
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CD177 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CD177 in Western blot, you can use appendix tissue lysate.
Positive control: Appendix (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: CD177 is membrane associated and extracellular, so signal in whole-tissue lysate may be weak.
HPA tissue expression evidence for CD177
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 | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | High | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CD177 Western Blot Tips
Deeper troubleshooting and optimisation questions for CD177, answered from its protein features.
How should CD177 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 CD177 isoforms could produce different bands?
Isoforms · UniProt lists isoforms 1, 2, and 3. Isoform 2 replaces residues 210–248 and lacks 249–437; isoform 3 replaces 129–145 and lacks 146–437. Both shortened sequences could differ in size from isoform 1. Check which isoforms the sample and antibody can detect before assigning a band.
Only if the antibody recognizes a region present in all three isoforms and the bands cannot be resolved separately. Isoform 3 lacks UniProt residues 146–437, while isoform 2 lacks 249–437 and has a different sequence at 210–248. Report the antibody region and which bands were included when presenting a total CD177 measurement.
Only if the antibody recognizes a region present in all three isoforms and the bands cannot be resolved separately. Isoform 3 lacks UniProt residues 146–437, while isoform 2 lacks 249–437 and has a different sequence at 210–248. Report the antibody region and which bands were included when presenting a total CD177 measurement.
How can CD177 glycosylation affect band interpretation?
PTM · UniProt lists one N-linked glycosylation site at Asn189. That coordinate is present in isoforms 1 and 2 and absent from isoform 3. A band difference alone cannot establish whether Asn189 is occupied or whether glycosylation caused the difference; compare appropriately treated and untreated samples if testing that explanation.
Does this guide establish induction of CD177?
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 CD177?
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 A03958-1 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.
What sample fraction is relevant for CD177 quantitation?
Quantitation · CD177 is annotated at the cell membrane and has a GPI-anchor keyword. Keep the sampled fraction consistent across lanes and account for CD177 that may be outside that fraction. If the antibody detects multiple isoforms, specify whether quantitation uses one band or their combined signal.
Why might CD177 migrate differently from its predicted 46.4 kDa?
Interpretation · The 46.4 kDa prediction does not account for every processed form. UniProt lists a signal peptide at residues 1–21, a propeptide at 409–437, and N-linked glycosylation at Asn189. These features can affect molecular mass, but they do not establish a visible shift or a specific apparent band size. Compare any observed band with the antibody’s target region and the sample preparation.
Match its binding region to the isoforms of interest. A region within UniProt residues 249–437 is absent from isoforms 2 and 3. Residues 146–209 are absent from isoform 3 but retained in isoform 2; residues 210–248 differ in isoform 2. Confirm the antibody’s numbering convention before comparing its stated region with UniProt coordinates.
First compare their apparent sizes and the antibody’s binding region with isoforms 2 and 3, which lack residues 249–437 and 146–437, respectively. Consider the annotated signal peptide at 1–21 and propeptide at 409–437 when assessing processing. These features identify possibilities; band position alone cannot establish identity.
Match its binding region to the isoforms of interest. A region within UniProt residues 249–437 is absent from isoforms 2 and 3. Residues 146–209 are absent from isoform 3 but retained in isoform 2; residues 210–248 differ in isoform 2. Confirm the antibody’s numbering convention before comparing its stated region with UniProt coordinates.
First compare their apparent sizes and the antibody’s binding region with isoforms 2 and 3, which lack residues 249–437 and 146–437, respectively. Consider the annotated signal peptide at 1–21 and propeptide at 409–437 when assessing processing. These features identify possibilities; band position alone cannot establish identity.
Boster reagents
CD177 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-CD177 antigen CD177 Antibody
Cat # A03958-1
Real WB data
Anti-CD177 antigen CD177 Antibody
Cat # A03958
Two the supplier anti-CD177 antibodies, A03958-1 and A03958, list human and mouse reactivity. Each has a Western blot image: mouse bone marrow for A03958-1 and K562 cells for A03958. These examples document specific tested contexts, not broader validation.
Which to pick: Choose A03958-1 if the mouse bone marrow blot and its reported conditions best match your experiment; choose A03958 if the K562 blot is more relevant. Both list human and mouse reactivity and have WB images.
Source: BosterBio CD177 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8N6Q3.
- Human Protein Atlas. CD177 tissue expression.
- PMC4057222 — target-verified WB comparison
- PMC10931634 — target-verified WB comparison
- PMC5502425 — target-verified WB comparison