APLP2 / Amyloid beta precursor like protein 2 · IHC design guide

Design Immunohistochemistry for APLP2

Plan APLP2 chromogenic IHC on paraffin sections using the 2–5 μg/ml catalog antibody range (datasheet A01232-1). Compare granular cytoplasmic staining in adipocytes and adrenal glandular cells with the tissue reference, and assess membrane or nuclear signal separately (HPA tissue IHC; UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for APLP2 (IHC for APLP2): expected localisation Granular cytoplasm (HPA tissue IHC); membrane and nucleus annotated (UniProt), antibody A01232-1, validated IHC image, and IHC protocol steps — Printable **APLP2** IHC protocol sheet — expected localisation Granular cytoplasm (HPA tissue IHC); membrane and nucleus annotated (UniProt), antibody A01232-1, controls and protocol steps. Open the full APLP2 IHC guide →

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APLP2 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	Granular cytoplasm (HPA tissue IHC); membrane and nucleus annotated (UniProt)
	Staining pattern	Granular cytoplasmic staining in most tissues (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A01232-1)
	Positive control ⓘ	Adipose tissue+4 more · see all
	Negative control ⓘ	None in HPA (detected in all 45 tissues); use no-primary + isotype controls

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep formalin fixation consistent across sections. (standard IHC practice; not target-specific)
	Caveat	Soluble APLP2 may complicate membrane-only scoring (UniProt)
	Regulation	Staining regulation is not established (UniProt)
	Isoform / epitope	6 isoforms; check whether the epitope is extracellular or cytoplasmic (UniProt)

Section 1

Recommended APLP2 IHC & IF Protocols

The catalog antibody has a datasheet IHC-P protocol (datasheet: A01232-1). The published protocols below describe retinal sections (PMC4897877) and human tumor microarrays and mouse tumors (PMC4385836).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A01232-1)
Fixation	Image fixative and duration unreported (datasheet A01232-1); 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 A01232-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A01232-1)
Primary antibody	Rabbit anti-APLP2, 2-5 μg/ml (datasheet A01232-1)
Primary incubation	Overnight at 4 °C (datasheet A01232-1)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A01232-1)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	APLP2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in most tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A01232-1); citrate at pH 6.8 is a published alternative for human tumor microarrays (PMC4385836).

Section 2

What Is the Expected APLP2 Staining Pattern?

APLP2 should appear as granular cytoplasmic staining across many tissues, including strong staining in selected adipocytes, glandular cells, respiratory epithelial cells, hematopoietic cells and neurons (HPA tissue IHC: Approved; medium consistency). Membrane-associated signal is biologically plausible because APLP2 has an extracellular region and one transmembrane segment (UniProt Q06481 topology). HPA also reports Golgi and vesicle localization by ICC-IF (HPA subcellular).

What am I looking at on my slide?
Granular cytoplasmic staining in adipocytes or respiratory epithelial cells.	This matches the broad cytoplasmic pattern and the High staining reported for those cells; assess signal against adjacent unstained areas on the same slide (HPA tissue IHC; general IHC practice).
Predominantly nuclear or smooth, uniform staining with little cytoplasmic granularity.	This differs from the reported tissue IHC pattern (HPA tissue IHC). Nuclear localization is listed by UniProt, so location alone cannot prove an artefact; compare morphology and controls before interpreting it (UniProt Q06481 subcellular location; general IHC practice).
Strong staining in cholangiocytes or ovarian stroma cells.	HPA reports Low staining in these cells, so strong signal merits a control check; broad APLP2 expression means cell identity alone cannot establish cross-reactivity (HPA tissue IHC; general IHC practice).
Diffuse chromogen across tissue, extracellular spaces or the entire section.	A noncellular haze does not match granular cytoplasmic staining (HPA tissue IHC). Review blocking, washes and endogenous detection activity before scoring it as specific signal (general IHC practice).
No staining in a section containing a reported high-staining cell population.	An absent signal conflicts with the HPA example for that population, but is inconclusive without a working positive control; check staining conditions and antibody use (HPA tissue IHC; general IHC practice).

💡Expected APLP2 appearanceCall positive a cellular, granular cytoplasmic signal in the relevant cell population, with High staining possible in HPA-listed examples; diffuse noncellular chromogen or equally strong staining in HPA-listed low-staining cells calls for control review (HPA tissue IHC; general IHC practice).

How each factor affects the staining
Tissue and cell selection	HPA describes expression in most tissues, with High examples in adipocytes, glandular cells, hematopoietic cells and selected neural cells; cholangiocytes and ovarian stroma cells are Low examples, not established negatives (HPA tissue IHC).
Epitope location and processing	APLP2 has an extracellular region at residues 32–692 and a cytoplasmic region at 717–763; UniProt also describes a soluble form. Without the antibody epitope, these facts do not predict which cellular or shed forms the stain detects (UniProt Q06481 topology and function).
Isoforms and modifications	UniProt lists 6 isoforms and glycosylation at residue 626. Coverage of isoforms or modified protein cannot be inferred without epitope information (UniProt Q06481 isoforms and PTMs).
Antibody validation	HPA rates tissue IHC Approved with medium consistency between staining and RNA data; antibody HPA039319 is IHC Approved. Treat the atlas pattern as a reference, while checking specificity in the experiment (HPA tissue IHC; HPA antibody validation; general IHC practice).
IF/ICC: what localization is expected?	Golgi and vesicle staining is reported as approved in ICC-IF, which can resolve structures seen as cytoplasmic granules in tissue IHC; the IF result does not prescribe a paraffin IHC protocol (HPA subcellular; HPA tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Reported high-staining cells show no signal.	The stain may have failed, or retrieval and dilution may be unsuitable for this assay; no APLP2-specific retrieval sensitivity is established here (HPA tissue IHC; general IHC practice).	Run a reported high-staining cell population as a positive control, then check antibody dilution, detection and retrieval against the antibody's IHC instructions (HPA tissue IHC; general IHC practice).
Chromogen spreads beyond cells or obscures tissue structure.	Excess detection signal, incomplete washing or endogenous enzyme activity can produce background (general IHC practice).	Inspect a negative reagent control; review washes, blocking and endogenous activity suppression appropriate to the chromogen system (general IHC practice).
Strong staining appears in cholangiocytes or ovarian stroma.	These are Low HPA examples; strong signal could reflect assay background or a sample-specific result (HPA tissue IHC; general IHC practice).	Compare a negative reagent control and a reported high-staining population on the same run; score cellular localization before assigning specificity (HPA tissue IHC; general IHC practice).
Only nuclei stain strongly.	The distribution differs from HPA's granular cytoplasmic tissue profile, although UniProt lists a nuclear location (HPA tissue IHC; UniProt Q06481 subcellular location).	Check nuclear counterstain and negative control, then document whether reproducible cellular signal also appears outside nuclei (general IHC practice).
Membrane rims dominate with few cytoplasmic granules.	A membrane component is plausible from APLP2 topology, but HPA describes the tissue IHC pattern as granular cytoplasmic (UniProt Q06481 topology; HPA tissue IHC).	Record both compartments, inspect a reported high-staining cell population and controls, and avoid assigning the antibody's epitope without evidence (HPA tissue IHC; general IHC practice).
Staining varies across a section or between runs.	Uneven reagent coverage, section quality or detection conditions can affect chromogenic IHC (general IHC practice).	Compare controls across runs, inspect section integrity and reagent coverage, and score only interpretable cells with their compartment recorded (general IHC practice).

Source: UniProt Q06481 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing APLP2 staining

IHC analysis of APLP2 using anti-APLP2 antibody (A01232-1). APLP2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-APLP2 Antibody (A01232-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A01232-1 · IHCIHC analysis of APLP2 using anti-APLP2 antibody (A01232-1). APLP2 was detected in a paraffin-embedded section of human colorectal adenocarci

IHC analysis of APLP2 using anti-APLP2 antibody (A01232-1). APLP2 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-APLP2 Antibody (A01232-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A01232-1 · IHCIHC analysis of APLP2 using anti-APLP2 antibody (A01232-1). APLP2 was detected in a paraffin-embedded section of human liver cancer tissue.

IF analysis of APLP2 using anti-APLP2 antibody (A01232-1). APLP2 was detected in an immunocytochemical section of HELA cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-APLP2 Antibody (A01232-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A01232-1 · IFIF analysis of APLP2 using anti-APLP2 antibody (A01232-1). APLP2 was detected in an immunocytochemical section of HELA cells. Enzyme antigen

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000084234-APLP2/tissue/cerebral+cortex#imid_9950984

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000084234-APLP2/tissue/colon#imid_9950910

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000084234-APLP2/tissue/liver#imid_9950911

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000084234-APLP2/tissue/kidney#imid_9950912

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA039319; green = APLP2, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for APLP2 IHC & IF

🧪Run adipose tissue first and score adipocytes, which show high APLP2 staining (HPA: High in adipocytes). HPA detects APLP2 in all 45 scored tissues, so use no-primary and isotype controls for negative staining; any cells on the positive slide considered internal negatives should retain counterstain without specific DAB signal, but the supplied HPA rows do not establish an internal negative cell type (HPA: no negative tissue rows; standard IHC practice).

Positive control tissue: Adipose tissue (Adipocytes, HPA High)

Negative control tissue: None in HPA: APLP2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show APLP2 in A-431, U-251MG, U2OS, with annotated localisation: Golgi apparatus (approved), Vesicles (approved) (HPA subcellular).

Technical controls: Include a secondary-only slide, a matched nonimmune rabbit IgG isotype control, and a biological specificity control using APLP2 knockout material or validated antigen-peptide competition (caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase and check adipose sections for nonspecific DAB background (standard chromogenic IHC practice).

⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A01232-1 paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The reported IHC conditions include heat retrieval in EDTA at pH 8.0, but the supplied evidence does not establish whether retrieval is required or whether frozen sections or IF would be easier (selected-SKU caption: EDTA retrieval). In adipose tissue, score staining in the thin adipocyte cell rim rather than the clear lipid space (HPA: High in adipocytes; standard histology practice).

HPA tissue IHC evidence for APLP2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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
Adipose tissue	Adipocytes	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	High	Protein (IHC)	HPA →
Appendix	Glandular cells	High	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	High	Protein (IHC)	HPA →
Breast	Adipocytes	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
None in HPA: APLP2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.

Section 3

Advanced APLP2 IHC Tips

Troubleshoot APLP2 staining in paraffin sections by checking retrieval, detection controls and cellular localisation (datasheet A01232-1; HPA tissue IHC).

What retrieval conditions should I try first when APLP2 staining is weak?

Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01232-1). The selected antibody detected APLP2 in a paraffin section of human colorectal adenocarcinoma after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (caption A01232-1). Check heating consistency, cooling and section retention before changing the retrieval buffer, because uneven processing can produce patchy chromogenic staining (standard IHC practice). If staining remains weak, compare a small retrieval time series with identical detection settings and a reference section; assess tissue integrity alongside signal (standard IHC practice).

Could fixation explain weak or uneven APLP2 staining?

The A01232-1 caption describes a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (caption A01232-1). Record the fixative, fixation duration, processing history and section age for each specimen before comparing staining intensity (standard IHC practice). Test affected and reference sections together using EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C (datasheet A01232-1; caption A01232-1). If signal varies across batches, compare morphology and staining under matched processing conditions; those observations can identify a processing association without establishing an APLP2-specific fixation effect (standard IHC practice).

How should I assess granular cytoplasmic staining when APLP2 is a membrane protein?

Score granular cytoplasmic staining as a plausible APLP2 pattern in tissue, since it is reported across most tissues (HPA tissue IHC profile). APLP2 also has a transmembrane segment at residues 693–716, with extracellular residues 32–692 and cytoplasmic residues 717–763 (UniProt Q06481 topology). Golgi and vesicle localisation in cell imaging offers a further reason to inspect intracellular puncta, while UniProt also lists cell membrane and nucleus (HPA subcellular; UniProt Q06481 subcellular). Compare staining within identifiable cells, including distribution and morphology, against matched controls; compartment alone cannot establish antibody specificity (standard IHC practice).

Could isoforms or epitope position change what the IHC stain detects?

APLP2 has 6 listed isoforms, so interpret staining as antibody-reactive APLP2 rather than assigning it to one isoform without epitope evidence (UniProt Q06481 isoforms). The record places E1 at residues 46–205, the Kunitz region at 306–364, E2 at 373–564 and the membrane segment at 693–716 (UniProt Q06481 domains and topology). Its signal peptide spans residues 1–31, and a glycosylation site is listed at 626 (UniProt Q06481 processing and glycosylation). Check the antibody’s documented immunogen or epitope before attributing nuclear, granular or surface staining to a particular domain, modification or isoform (standard IHC interpretation).

How can I check an APLP2 IHC pattern with multiplex immunofluorescence?

Choose a second marker that identifies the cell population you are scoring, then compare APLP2 signal within those marked cells against adjacent unmarked cells (standard IF practice). For an adipose specimen, adipocyte staining provides a documented tissue comparison, while granular cytoplasmic expression is the broader IHC pattern (HPA tissue IHC: adipocytes High; HPA tissue IHC profile). Select fluorophores after examining tissue autofluorescence and include single-channel and no-primary controls to assess bleed-through and nonspecific signal (standard IF practice). Set permeabilisation according to the antibody’s documented epitope: extracellular residues 32–692 and cytoplasmic residues 717–763 lie on opposite sides of the membrane (UniProt Q06481 topology; standard IF practice).

What should I change when the DAB stain is widespread or dirty?

First compare the stained section with a no-primary control and inspect pigment, folds, edges and damaged regions before calling diffuse color APLP2 signal (standard IHC practice). The selected demonstration used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (caption A01232-1). Apply a peroxidase block and control DAB development time as general chromogenic IHC steps, especially if the no-primary section also colors (standard IHC practice). If background persists, compare primary concentration and washing conditions systematically while retaining the same reference section and retrieval conditions (standard IHC practice).

How should I quantify APLP2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and region of interest before scoring, because APLP2 staining can be granular and cytoplasmic across tissues (HPA tissue IHC profile; standard IHC practice). Report the percentage of positive eligible cells and an intensity distribution, or calculate an H-score from percentages at intensity levels 0–3, giving a possible range of 0–300 (standard IHC scoring practice). If counting positive cells per mm², also report eligible cell density so tissue cellularity does not masquerade as expression change (standard IHC quantification practice). Normalize comparisons to the same cell type, sampled area, retrieval, staining run and scoring thresholds, with a reference section on each run (standard IHC practice).

How can I distinguish credible APLP2 staining from artefact?

Give greatest weight to reproducible staining within intact, identifiable cells that matches a documented granular cytoplasmic pattern (HPA tissue IHC profile; standard IHC interpretation). APLP2 is also listed at the cell membrane and nucleus, and cell imaging places it in Golgi and vesicles, so an unexpected compartment merits control checks rather than automatic dismissal (UniProt Q06481 subcellular; HPA subcellular). Examine edges, necrotic regions and no-primary controls for nonspecific DAB deposition or endogenous peroxidase activity before interpreting isolated hotspots (standard IHC practice). Compare cell identity and tissue context with the documented staining profile; HPA rates the tissue IHC evidence Approved with medium consistency against RNA expression (HPA tissue IHC).

Boster reagents

Best APLP2 / Amyloid beta precursor like protein 2 IHC Antibodies

Image-documented APLP2 IHC covers human paraffin sections, and IF/ICC covers HELA cells with A01232-1; both antibodies list human, mouse, and rat reactivity (A01232-1 image captions; catalog reactivity).

Real IHC data

Anti-APLP2 Antibody ®

Cat # A01232-1

View product →

A01232-1 lists IHC and IF/ICC, with captions showing human colorectal adenocarcinoma, liver cancer, lung cancer, and testicular germ cell tumor paraffin sections, plus HELA cell IF (catalog applications; A01232-1 image captions). M01232 lists IHC and human, mouse, and rat reactivity, but has no supplied IHC or IF image caption (catalog applications/reactivity; M01232 image alts).

Which to pick: For tissue IHC, choose A01232-1 for its documented paraffin-section workflow: its IHC caption reports EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (A01232-1 IHC caption). For IF/ICC, choose A01232-1 at the listed 5 μg/ml; M01232 does not list IF/ICC (catalog applications/dilutions). For cross-species IHC planning, both list human, mouse, and rat reactivity; M01232 is rabbit monoclonal clone 25A47 without a supplied IHC image, while A01232-1 has human paraffin-section images (catalog host/clone/reactivity; IHC image captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q06481 (APLP2_HUMAN, Amyloid beta precursor like protein 2).
Human Protein Atlas. APLP2 tissue IHC expression (reliability: Approved).
Human Protein Atlas. APLP2 subcellular location (ICC-IF): Localized to the Golgi apparatus and vesicles..
Human Protein Atlas. APLP2 antibody validation summary (1 antibodies).
Amyloid Precursor-like Protein 2 Expression Increases during Pancreatic Cancer Development and Shortens the Survival of a Spontaneous Mouse Model of Pancreatic Cancer. Cancers 2021 — PMC8036577.
Amyloid Precursor-Like Protein 2 deletion-induced retinal synaptopathy related to congenital stationary night blindness: structural, functional and molecular characteristics. Molecular brain 2016 — PMC4897877.
APLP2 Regulates Refractive Error and Myopia Development in Mice and Humans. PLoS genetics 2015 — PMC4551475.
Amyloid precursor-like protein 2 (APLP2) affects the actin cytoskeleton and increases pancreatic cancer growth and metastasis. Oncotarget 2015 — PMC4385836.
PubMed PMID:8485127 — UniProt-cited evidence.
PubMed PMID:8220435 — UniProt-cited evidence.
PubMed PMID:7702756 — UniProt-cited evidence.