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GP1BA / Platelet glycoprotein Ib alpha chain · IHC design guide
Design Immunohistochemistry for GP1BA
This GP1BA IHC-P guide covers selective cytoplasmic staining in platelets and megakaryocytes (HPA tissue IHC). It gives a catalog antibody starting dilution of 1:50 (datasheet A02073-4) and flags the glycocalicin fragment when choosing an epitope (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GP1BA Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Platelet cytoplasm observed; membrane expected (HPA tissue IHC; UniProt) | |
| Staining pattern | Selective cytoplasmic staining in platelets and megakaryocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02073-4) | |
| Positive control | Bone marrow+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Glycocalicin (17–506) may affect extracellular epitope staining (UniProt) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | No isoforms; glycocalicin lacks the cytoplasmic tail (UniProt) |
Section 1
Recommended GP1BA IHC & IF Protocols
The IHC-validated antibody’s paraffin-section protocol (datasheet: A02073-4) is accompanied by published GP1BA IHC protocols for cardiac and airway tissues (PMC12984341) and tumor tissue (PMC13397375).
| Sample | Paraffin-embedded mouse spleen tissue; fixative not specified (datasheet A02073-4) |
| Fixation | Image fixative and duration unreported (datasheet A02073-4); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A02073-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02073-4) |
| Primary antibody | Rabbit anti-GP1BA, 1:50 recommended; image 1:100 (datasheet A02073-4) |
| Primary incubation | Overnight at 4 °C (datasheet A02073-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02073-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GP1BA-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: Medium). HPA tissue profile: Selective cytoplasmic expression in platelets and megakaryocytes. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A02073-4); consider citrate pH 6.0 as used in both published protocols (PMC12984341; PMC13397375).
Section 2
What Is the Expected GP1BA Staining Pattern?
GP1BA is a platelet surface membrane protein with an extracellular region at residues 17–531, a transmembrane segment at 532–552, and a cytoplasmic tail at 553–652 (UniProt P07359 topology). In tissue IHC, expect selective staining of platelets and megakaryocytes; HPA describes the observed signal as cytoplasmic and rates its tissue profile Supported, citing consistency between staining and RNA expression (HPA tissue IHC).
| Selective staining of platelets and megakaryocytes, with signal appearing cytoplasmic in tissue sections (HPA tissue IHC). | This matches HPA’s reported pattern. The cytoplasmic appearance in IHC does not overturn GP1BA’s membrane topology; the two descriptions refer to observed tissue staining and protein location, respectively (HPA tissue IHC; UniProt P07359 topology). |
| Predominantly nuclear staining in cells scored as positive. | Nuclear localisation is inconsistent with the membrane location assigned to GP1BA (UniProt P07359). Review morphology, counterstain and detection controls before scoring this as target signal (general IHC practice). |
| Strong staining of adipocytes or respiratory epithelial cells. | These cell types were reported as Not detected in the sampled adipose tissue and bronchus, respectively (HPA tissue IHC). Consider nonspecific binding or endogenous detection activity; a control can help distinguish them (general IHC practice). |
| Diffuse colour across cells and surrounding tissue, obscuring platelet or megakaryocyte outlines. | This is difficult to reconcile with HPA’s selective cellular pattern (HPA tissue IHC). Inspect the no-primary control, blocking, washing and chromogen development before interpreting any cell as positive (general IHC practice). |
| No convincing signal in an otherwise interpretable bone marrow section. | HPA reports Medium staining in bone marrow hematopoietic cells and identifies platelets and megakaryocytes in its selective profile (HPA tissue IHC). Confirm that those elements are present, then check the assay controls and section processing (general IHC practice). |
💡Expected GP1BA appearanceCall positive when platelets or megakaryocytes show selective cytoplasmic-appearing IHC signal, with Medium staining reported for bone marrow hematopoietic cells and splenic red-pulp cells; diffuse staining or staining dominated by unrelated cells is suspect (HPA tissue IHC).
| Tissue and cell selection (HPA tissue IHC) | Bone marrow hematopoietic cells and splenic red-pulp cells are listed at Medium staining, while the profile specifically identifies platelets and megakaryocytes. Interpret those tissue-level entries alongside cell morphology; they do not mean every hematopoietic or red-pulp cell expresses GP1BA (HPA tissue IHC). |
| Membrane topology and observed IHC pattern (UniProt P07359; HPA tissue IHC) | The extracellular region spans residues 17–531, followed by one transmembrane segment and a cytoplasmic tail (UniProt P07359 topology). HPA nevertheless calls its tissue staining cytoplasmic. Judge concordance against the reported slide pattern without claiming that this determines the antibody’s epitope (HPA tissue IHC). |
| Protein processing and epitope uncertainty (UniProt P07359) | UniProt annotates a full chain at residues 17–652 and glycocalicin at residues 17–506. An antibody’s exact epitope is not supplied here, so this record cannot establish whether it detects both products or whether processing explains a particular tissue pattern (UniProt P07359 processing). |
| Antibody evidence (HPA antibody validation) | HPA lists two IHC antibodies, HPA013316 and CAB002496, each with Supported status; neither has an ICC status in the supplied entries (HPA antibody validation). Supported tissue evidence helps anchor interpretation, but it is not a validation result for a different catalog antibody (HPA antibody validation). |
| IF/ICC question: is the tissue IHC pattern an IF/ICC reference? (HPA subcellular; HPA antibody validation) | HPA summarizes the subcellular location as Membrane but supplies no main location or ICC-IF images, and neither listed antibody has an ICC status (HPA subcellular; HPA antibody validation). Use the separate IF/ICC guide for assay design; this record alone cannot establish an expected image pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow lacks visible positive elements or shows no signal (HPA tissue IHC). | The sampled area may lack identifiable platelets or megakaryocytes, or the assay may have failed; HPA’s Medium bone marrow entry alone cannot separate these explanations (HPA tissue IHC). | Check cell morphology and section quality, then review the positive control, primary-antibody step, retrieval record and detection reagents (general IHC practice). |
| Only nuclei appear positive. | This differs from the reported cytoplasmic tissue pattern and membrane location (HPA tissue IHC; UniProt P07359). Counterstain or detection artefact is possible (general IHC practice). | Compare the no-primary control and counterstain, and inspect whether any platelet or megakaryocyte-associated signal remains (general IHC practice). |
| Adipocytes or bronchial epithelium stain prominently. | HPA reports Not detected in adipocytes from adipose tissue and respiratory epithelial cells from bronchus (HPA tissue IHC). Nonspecific or endogenous detection signal is possible (general IHC practice). | Run a no-primary control and review blocking and detection conditions; score only signal that also fits cell morphology and the expected tissue distribution (general IHC practice; HPA tissue IHC). |
| Brown signal is widespread and obscures cell boundaries. | Diffuse background prevents assessment of HPA’s selective platelet and megakaryocyte pattern (HPA tissue IHC). Excess detection signal or inadequate washing can contribute (general IHC practice). | Compare a no-primary section, check wash steps and chromogen development, and reassess only areas with interpretable cell boundaries (general IHC practice). |
| A positive cell looks cytoplasmic rather than sharply outlined at its membrane. | HPA expressly describes cytoplasmic expression in tissue IHC, while UniProt assigns GP1BA to the membrane (HPA tissue IHC; UniProt P07359). | Assess the cell type and selective distribution first. Do not reject a plausible tissue IHC result solely because its edge is indistinct (HPA tissue IHC). |
| IF/ICC shows an unexpected distribution. | The supplied HPA subcellular entry has no ICC-IF images, and both listed antibodies lack ICC status (HPA subcellular; HPA antibody validation). | Treat the IF/ICC pattern as unconfirmed by this tissue IHC record and consult the separate IF/ICC guide; compare localisation with membrane topology using assay controls (UniProt P07359; general IF practice). |
Experiment evidence
IHC & IF images showing GP1BA staining
A02073-4 · IHCIHC analysis of CD42b/GP1BA using anti-CD42b/GP1BA antibody (A02073-4). CD42b/GP1BA was detected in a paraffin-embedded section of mouse spl
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000185245-GP1BA/tissue/cerebral+cortex#imid_774624
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000185245-GP1BA/tissue/colon#imid_774550
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000185245-GP1BA/tissue/liver#imid_774559
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000185245-GP1BA/tissue/kidney#imid_774552
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GP1BA IHC & IF
🧪Run spleen first and look for staining in red-pulp cells (HPA: Medium in cells in red pulp). Use bronchus respiratory epithelial cells as the negative tissue (HPA: Not detected); on the spleen slide, cells outside the stained red-pulp compartment should provide an internal negative comparison, allowing for platelet-associated signal (UniProt P07359 function).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for GP1BA; derive a cell-line control from the positive tissue's cell type (Hematopoietic cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a rabbit IgG isotype control matched to the primary antibody (selected-SKU caption: rabbit primary and anti-rabbit secondary); use GP1BA knockout tissue, if available, as a biological negative. Quench endogenous peroxidase and assess blood-associated background in spleen before interpreting DAB staining (selected-SKU caption: spleen section with HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU caption does not state the fixative (selected-SKU caption: fixative not stated). Paraffin-section staining is illustrated after heat retrieval in EDTA at pH 8.0, but the caption does not establish that retrieval is required (selected-SKU caption: EDTA retrieval, pH 8.0). The supplied evidence does not establish whether frozen sections or IF are easier; interpret spleen staining with care because blood-associated background can complicate chromogenic detection (selected-SKU caption: spleen HRP/DAB staining).
HPA tissue IHC evidence for GP1BA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended 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 → |
| Appendix | Glandular 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 GP1BA IHC Tips
Troubleshoot GP1BA staining in paraffin sections by checking retrieval, cell identity, membrane localisation and assay controls before comparing chromogenic signal across samples.
How should I retrieve GP1BA in paraffin sections with weak staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A02073-4). The illustrated paraffin-section workflow used this retrieval before 10% goat-serum blocking and primary antibody incubation overnight at 4°C (datasheet A02073-4). If staining remains weak, compare a shorter and longer heating interval on matched sections while keeping detection conditions fixed (standard IHC practice). Score signal in platelet and megakaryocyte-rich areas alongside section damage and background; these are the expected positive cell types, while excessive heating can impair morphology (HPA tissue IHC; standard IHC practice).
Could fixation explain inconsistent GP1BA staining between paraffin blocks?
The selected paraffin-section image does not state its fixative, so target-specific GP1BA fixation sensitivity is unknown (datasheet A02073-4). Record each block’s fixative and fixation duration, then compare matched sections using the same EDTA pH 8.0 retrieval and detection workflow (datasheet A02073-4; standard IHC practice). Use a control section with identifiable platelets or megakaryocytes in each staining run, because these are the reported positive cell types (HPA tissue IHC; standard IHC practice). If signal differs between blocks, inspect morphology and processing records before attributing the difference to biology; fixation can alter antigen accessibility in paraffin IHC (standard IHC practice).
Should GP1BA stain the membrane or cytoplasm in tissue sections?
GP1BA is a membrane protein with an extracellular region at residues 17–531, a transmembrane segment at 532–552 and a cytoplasmic region at 553–652 (UniProt P07359 topology). HPA describes selective cytoplasmic staining in platelets and megakaryocytes, so assess the stained cell and its morphology before treating an apparently cytoplasmic chromogenic pattern as discordant (HPA tissue IHC; standard IHC practice). In thin paraffin sections, membrane-associated signal may be difficult to separate visually from the small platelet profile (standard IHC practice). Diffuse staining across unrelated cell populations warrants review of the negative control and background before assigning GP1BA localisation (HPA tissue IHC; standard IHC practice).
How might GP1BA processing affect interpretation of an unknown antibody epitope?
The record lists 0 isoforms and describes a full chain spanning residues 17–652 plus glycocalicin spanning 17–506 (UniProt P07359 processing and isoforms). An antibody recognising the shared extracellular sequence could detect either form, whereas an epitope in the cytoplasmic region would distinguish the membrane-associated chain from glycocalicin (UniProt P07359 topology and processing). The supplied caption does not identify the antibody epitope, so neither outcome can be assigned to A02073-4 without epitope information (datasheet A02073-4). GP1BA also has 44 annotated glycosylation sites; compare retrieval conditions empirically if accessibility is uncertain, without assuming which modification affects this antibody (UniProt P07359 glycosylation; standard IHC practice).
How can I investigate GP1BA by IF alongside chromogenic IHC?
Treat IF as a separate assay requiring its own controls; the supplied A02073-4 image documents paraffin-section chromogenic IHC, with no IF result provided (datasheet A02073-4). Multiplex GP1BA with an independently validated platelet or megakaryocyte marker such as CD41, and check overlap at the cell level (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore when tissue autofluorescence interferes with shorter wavelengths, and include unstained and single-label controls (standard IF practice). Adjust permeabilisation only after establishing whether the antibody recognises the extracellular 17–531 or cytoplasmic 553–652 region; the supplied caption does not specify its epitope (UniProt P07359 topology; datasheet A02073-4; standard IF practice).
What should I check when GP1BA DAB staining looks diffuse?
First compare the stained section with a no-primary control to assess secondary-reagent signal and endogenous peroxidase activity (standard IHC practice). The illustrated workflow used 10% goat serum, primary antibody at 1:100 overnight at 4°C, and peroxidase-linked detection with DAB (datasheet A02073-4). A peroxidase block, appropriate washes and a controlled DAB development interval are general ways to reduce chromogenic background (standard IHC practice). If diffuse colour persists, compare it with the expected platelet and megakaryocyte distribution and with areas of tissue damage before interpreting it as GP1BA (HPA tissue IHC; standard IHC practice).
How should I quantify GP1BA staining without confusing abundance and area? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine a scoring unit: percent positive platelets or megakaryocytes, an intensity-weighted H-score for identifiable megakaryocytes, or positive-object density per mm² of evaluable tissue (HPA tissue IHC; standard IHC practice). Keep the threshold, counterstain, imaging settings and sampled area consistent across sections (standard IHC practice). For cell-based scores, normalise positive counts to the number of eligible cells; for density, normalise to measured tissue area and exclude folds or necrosis (standard IHC practice). Report platelet-rich and megakaryocyte-rich compartments separately when morphology permits, because HPA identifies both as positive and a combined score can obscure changes in their proportions (HPA tissue IHC; standard IHC practice).
How can I distinguish genuine GP1BA staining from artefact?
A convincing result identifies platelets or megakaryocytes in the stained structures, consistent with the reported selective pattern and GP1BA’s platelet surface role (HPA tissue IHC; UniProt P07359 function). In spleen, compare staining with red-pulp cell morphology; HPA reports medium staining in red-pulp cells, and the A02073-4 caption documents a mouse-spleen paraffin section (HPA tissue IHC; datasheet A02073-4). Review apparent staining in unrelated cells against a no-primary control, especially at section edges or necrotic areas, where artefacts can concentrate (HPA tissue IHC; standard IHC practice). Persistent colour in the no-primary control suggests detection background, including endogenous peroxidase, rather than evidence of GP1BA (standard IHC practice).
Boster reagents
Best GP1BA / Platelet glycoprotein Ib alpha chain IHC Antibodies
The catalog lists two IHC antibodies for GP1BA (catalog applications); only A02073-4 has an IHC image, from a paraffin-embedded mouse spleen section (A02073-4 image caption). Neither has IF/ICC data (catalog applications and images).
Real IHC data
Anti-CD42b/GP1BA Antibody
Cat # A02073-4
A02073-4 lists human and mouse reactivity and IHC use (catalog reactivity and applications); its image shows staining in a paraffin-embedded mouse spleen section (A02073-4 image caption). M02073-2 lists human reactivity and IHC use, with no IHC image supplied (catalog reactivity, applications and images).
Which to pick: Choose A02073-4 for tissue IHC when a demonstrated paraffin-section example matters; its mouse spleen caption reports antigen retrieval in EDTA at pH 8.0, but does not report the fixative (A02073-4 image caption). For a human-reactive monoclonal IHC option, M02073-2 is listed as clone 17G60, though no tissue image is supplied (M02073-2 catalog clone, applications, reactivity and images). For human-and-mouse reactivity, choose A02073-4 (catalog reactivity); neither SKU has listed IF/ICC validation, so no IF/ICC choice is supported here (catalog applications and images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P07359 (GP1BA_HUMAN, Platelet glycoprotein Ib alpha chain).
- Human Protein Atlas. GP1BA tissue IHC expression (reliability: Supported).
- Human Protein Atlas. GP1BA subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. GP1BA antibody validation summary (2 antibodies).
- Integrative Proteomic and Transcriptomic Profiling Identifies Candidate Biomarkers for Discriminating Anaphylactic from Cardiac Sudden Death. International journal of molecular sciences 2026 — PMC12984341.
- miR-320d Is Associated with Reduced Nasopharyngeal Carcinoma Progression, Potentially through the NF-κB/IL-8 Axis-Mediated Inhibition of Neutrophil Extracellular Trap Formation. Oncology research 2026 — PMC13397375.
- PubMed PMID:3303030 — UniProt-cited evidence.
- PubMed PMID:2845978 — UniProt-cited evidence.
- PubMed PMID:12038791 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GP1BA (Platelet glycoprotein Ib alpha chain). Expected localisation Platelet cytoplasm observed; membrane expected (HPA tissue IHC; UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02073-4). Validated IHC antibody: A02073-4 at 1:50 recommended; image 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse spleen tissue; fixative not specified (datasheet A02073-4); 2. Fixation: Image fixative and duration unreported (datasheet A02073-4); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A02073-4); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02073-4); 8. Primary antibody: Rabbit anti-GP1BA, 1:50 recommended; image 1:100 (datasheet A02073-4); 9. Primary incubation: Overnight at 4 °C (datasheet A02073-4). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Glycocalicin (17–506) may affect extracellular epitope staining (UniProt); Isoform / epitope: No isoforms; glycocalicin lacks the cytoplasmic tail (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt P07359, HPA tissue IHC · Supported, HPA ICC-IF · Membrane, Datasheet A02073-4, PMC12984341, PMC13397375. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GP1BA/immunohistochemistry-ihc-guide.