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- Table of Contents
Plan ABCA7 IHC in paraffin sections using tissue controls, consistent fixation and chromogenic detection. Bone marrow hematopoietic cells show high staining (HPA tissue IHC); the catalog antibody A03163 starts at 5 μg/mL for IHC (datasheet: A03163).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasm in bone marrow and immune cell subsets (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in marrow and immune cell subsets (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03163) | |
| Caveat | Presumed off-target binding may mimic positive staining (HPA tissue IHC) | |
| Regulation | Enriched in myelo-lymphatic tissues (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope coverage across extracellular and cytoplasmic regions (UniProt) |
The catalog antibody’s IHC-P protocol is paired with a published ABCA7 protocol for paraffin-embedded ovarian tumor specimens (PMC6176408).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A03163); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-ABCA7, 5 μg/mL (datasheet A03163) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ABCA7-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in bone marrow and in subsets of immune cells in several tissues. No signal in the no-primary control. |
ABCA7 is a 15-pass membrane protein with reported cell membrane, Golgi, endosomal and cytoplasmic locations (UniProt Q8IZY2 topology and subcellular location). In paraffin-section IHC, expect cytoplasmic staining in bone marrow hematopoietic cells and subsets of immune cells; ciliary rootlets can also stain strongly (HPA tissue IHC). HPA rates its tissue profile Enhanced, while noting medium consistency with RNA data and presumed off-target binding that was disregarded (HPA tissue IHC).
| Strong staining in bone marrow hematopoietic cells, concentrated within the cell body. | This fits HPA's High staining level and cytoplasmic tissue profile (HPA tissue IHC). Score the named cells and their compartment; a positive field alone does not establish that every cell is positive (HPA tissue IHC). |
| Distinct staining at ciliary rootlets in fallopian tube or nasopharynx. | Both are reported as High in ciliated cells specifically at ciliary rootlets (HPA tissue IHC). Record the restricted rootlet pattern rather than calling the entire ciliated cell uniformly positive (HPA tissue IHC). |
| Predominantly nuclear staining in otherwise expected positive cells. | A nuclear dominant pattern is discordant with the reported cytoplasmic tissue pattern and membrane-associated locations (HPA tissue IHC; UniProt Q8IZY2 subcellular location). Treat it as suspect and compare controls before scoring it as ABCA7 (standard IHC practice). |
| Strong staining in adipocytes across adipose tissue or breast. | HPA reports ABCA7 as Not detected in these adipocytes (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity; an unexpected positive cell type needs independent support before interpretation (HPA tissue IHC; standard IHC practice). |
| No detectable signal in bone marrow hematopoietic cells. | That conflicts with the reported High IHC level (HPA tissue IHC). First assess section quality, retrieval and detection with appropriate controls; a blank slide by itself cannot distinguish a technical failure from a specimen difference (standard IHC practice). |
| Cell and tissue selection | Bone marrow hematopoietic cells are High, splenic red-pulp cells Medium, and lung macrophages Low; use the reported cell population when judging a result (HPA tissue IHC). A weak lung macrophage signal should not be judged against the bone marrow level (HPA tissue IHC). |
| Compartment and cell state | Resting macrophages are reported mainly cytoplasmic, probably in Golgi and endosomes; stimulation with C1q or apoptotic cells can shift ABCA7 to membrane ruffles and phagocytic cups (UniProt Q8IZY2 subcellular location). Interpret a localized membrane pattern in that context, while using HPA's cytoplasmic tissue profile for routine IHC (HPA tissue IHC). |
| Topology and epitope location | ABCA7 has 15 transmembrane segments and two annotated extracellular regions (UniProt Q8IZY2 topology). The supplied record gives no antibody epitope, so it cannot predict this antibody's access after retrieval or permeabilisation (UniProt Q8IZY2 topology; supplied HPA antibody record). |
| Isoforms and protein processing | Two isoforms are annotated, and the supplied record lists one chain spanning residues 1–2146, with no signal peptide or propeptide annotation (UniProt Q8IZY2 isoforms and processing). Antibody recognition of either isoform is unspecified; do not infer isoform-specific staining or cleavage from these annotations (UniProt Q8IZY2; supplied HPA antibody record). |
| Strength of the IHC reference | HPA lists HPA041564 as IHC Enhanced, but its tissue summary reports medium agreement with RNA data and presumed off-target binding that was disregarded (HPA antibodies; HPA tissue IHC). Use the named positive cells and negative comparators together when assessing an unexpected pattern (HPA tissue IHC; standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow hematopoietic cells are blank. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Check tissue preservation, retrieval, primary-antibody incubation and chromogenic detection against run controls; repeat with an appropriate positive section if needed (standard IHC practice). |
| The whole section has diffuse brown background. | Nonspecific reagent binding, incomplete blocking or excessive chromogen development can obscure a cell-restricted pattern (standard IHC practice). | Compare a primary-omission control, review blocking and washes, and adjust detection development according to the assay controls (standard IHC practice). |
| Unexpected strong staining appears in adipocytes or another reported negative cell population. | Cross-reactivity or endogenous detection activity is possible; adipocytes in adipose tissue and breast are Not detected in HPA IHC (HPA tissue IHC; standard IHC practice). | Inspect primary-omission and relevant endogenous-activity controls, then compare the same run with bone marrow hematopoietic cells before assigning specificity (HPA tissue IHC; standard IHC practice). |
| The main signal is nuclear, with little staining in expected cytoplasmic cells. | The compartment disagrees with HPA's cytoplasmic tissue pattern and UniProt's membrane and organelle locations (HPA tissue IHC; UniProt Q8IZY2 subcellular location). | Check counterstain and detection controls, then score nuclear signal separately as unexpected until independently supported (standard IHC practice). |
| Ciliated cells stain broadly, but the rootlets cannot be resolved. | HPA specifies High staining at ciliary rootlets in fallopian tube and nasopharynx; broad staining alone does not reproduce that localization (HPA tissue IHC). | Review morphology and a suitably resolved field before calling the pattern concordant; document the compartment actually visible (HPA tissue IHC; standard IHC practice). |
| Q: Should IF/ICC show the same pattern as paraffin-section IHC? | The assays use different sample preparations, and HPA reports supported plasma membrane, Golgi and additional cell-junction localization in ICC-IF (HPA subcellular). | A: Interpret IF/ICC on its own guide page using those supported compartments; do not require every IHC-positive cell to show an identical IF/ICC pattern (HPA subcellular; HPA tissue IHC). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Nasopharynx | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Medium | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Troubleshoot ABCA7 staining in paraffin sections by checking retrieval, cell type, compartment and controls before comparing signal across samples.
Both antibodies have human tissue IHC images; A03163 also has IF/ICC images in 293 cells and human spleen (catalog image captions). Listed reactivity spans human, mouse and rat (catalog reactivity).
A03163 renders with human spleen IHC at 5 μg/mL and has fluorescence images in 293 cells and human spleen (A03163 image captions). A03163-2 renders with paraffin-embedded human colon carcinoma IHC at 1:50, showing membrane staining and a PBS-for-primary control (A03163-2 IHC image caption).
Which to pick: For human tissue IHC, A03163 has an IHC-P listing and a human spleen image at 5 μg/mL (A03163 applications and IHC image caption). For paraffin sections, A03163-2 shows human colon carcinoma staining at 1:50; the fixative is unreported (A03163-2 IHC image caption). Choose A03163 for IF/ICC based on its application list and fluorescence images; A03163-2 lists mouse and rat reactivity for cross-species IHC, although its supplied IHC images show human tissue only (catalog applications, reactivity and image captions).