LRP8 / Low-density lipoprotein receptor-related protein 8 · IHC design guide

Design Immunohistochemistry for LRP8

Use testis pachytene spermatocytes as a high-staining reference (HPA tissue IHC), and start the IHC-validated antibody at 0.5–1 μg/mL (datasheet A03444-2). Account for secreted receptor fragments (UniProt) and disregard off-target nuclear staining (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for LRP8 (IHC for LRP8): expected localisation Membrane or secreted forms expected (UniProt); tissue compartment unreported (HPA tissue IHC), antibody A03444-2, validated IHC image, and IHC protocol steps — Printable **LRP8** IHC protocol sheet — expected localisation Membrane or secreted forms expected (UniProt); tissue compartment unreported (HPA tissue IHC), antibody A03444-2, controls and protocol steps. Open the full LRP8 IHC guide →

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LRP8 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	Membrane or secreted forms expected (UniProt); tissue compartment unreported (HPA tissue IHC)
	Staining pattern	High in testis pachytene spermatocytes; subcellular pattern unreported (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A03444-2)
	Positive control ⓘ	Testis
	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	Nuclear staining may be off-target (HPA tissue IHC)
	Regulation	No staining regulator specified in the record (UniProt)
	Isoform / epitope	5 isoforms; shedding makes extracellular versus cytoplasmic epitopes relevant (UniProt)

Section 1

Recommended LRP8 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet A03444-2) is followed by four published LRP8 IHC protocols (PMC12339728; PMC11954824; PMC12964802; PMC12412626).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A03444-2)
Fixation	Image fixative and duration unreported (datasheet A03444-2); 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 A03444-2); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A03444-2)
Primary antibody	Rabbit anti-LRP8, 0.5-1μg/ml (datasheet A03444-2)
Primary incubation	Overnight at 4 °C (datasheet A03444-2)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03444-2)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	LRP8-positive staining in pachytene spermatocytes of testis (HPA tissue IHC: High). HPA tissue profile: secreted protein expression. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A03444-2). Sodium citrate retrieval is published for another LRP8 antibody (PMC12412626).

Section 2

What Is the Expected LRP8 Staining Pattern?

LRP8 is a cell membrane receptor with an extracellular domain and cytoplasmic tail; some isoforms yield a secreted fragment (UniProt Q14114 topology and processing). In tissue IHC, pachytene spermatocytes in testis are the clearest supplied positive population, with High staining (HPA: testis IHC). HPA rates the tissue pattern Enhanced but disregards observed nuclear off target binding (HPA: reliability description).

What am I looking at on my slide?
Pachytene spermatocytes stain more strongly than nearby cells, with signal outlining cells.	This fits the supplied High testis cell population (HPA: pachytene spermatocytes) and a membrane receptor (UniProt Q14114 topology). Evaluate the cell population and compartment together; the evidence does not specify a uniform rim in every section.
Nuclei stain prominently, especially when cell boundaries are faint.	Treat nuclear signal cautiously: HPA observed nuclear off target binding and excluded it from its LRP8 interpretation (HPA: reliability description). Nuclear staining alone does not establish target detection, even in a tissue containing positive cells.
Adipocytes or adrenal glandular cells stain as strongly as the testis positive cells.	These specific cell populations were Not detected in HPA tissue IHC (HPA: adipocytes; adrenal glandular cells). Check cellular identification and controls before assigning the signal to LRP8; unexpected staining can reflect cross reactivity or detection background.
A broad brown haze covers cells, extracellular spaces, or the whole section.	Diffuse signal is difficult to assign to a compartment. A shed receptor fragment is biologically plausible (UniProt Q14114 processing), but a widespread haze alone cannot distinguish it from nonspecific chromogen deposition or incomplete washing (general IHC practice).
Pachytene spermatocytes show no convincing signal.	A negative result conflicts with the High staining reported for this population (HPA: testis IHC), but does not by itself prove LRP8 absence. Confirm the cells are present, then review the staining run and antibody performance (general IHC practice).

💡Expected LRP8 appearanceCall positive when pachytene spermatocytes show clearly stronger, plausibly cell associated staining consistent with a membrane receptor (HPA: High in pachytene spermatocytes; UniProt Q14114 topology); isolated nuclear staining is a false positive concern (HPA: nuclear off target binding).

How each factor affects the staining
Receptor orientation and epitope location	LRP8 spans the membrane at residues 827–847, with extracellular residues 42–826 and cytoplasmic residues 848–963 (UniProt Q14114 topology). The antibody epitope is not supplied, so topology cannot predict which receptor forms it detects.
Isoforms and receptor shedding	UniProt lists 5 isoforms; isoforms containing a furin type cleavage site can release a secreted receptor fragment (UniProt Q14114 isoforms and processing). Extracellular signal therefore needs context, and the supplied evidence cannot assign it to a particular isoform.
Interpretation across tissues	UniProt reports expression mainly in brain and placenta, while HPA reports High protein staining in testis pachytene spermatocytes and no detection in selected cerebellar granular layer cells (UniProt Q14114 tissue specificity; HPA: tissue IHC). These describe different scopes; do not score a whole organ from one listed cell population.
Antibody validation and compartment caveat	HPA073031 has Enhanced IHC validation (HPA: antibody validation), while HPA separately notes nuclear off target binding (HPA: reliability description). Enhanced validation supports use of the reported tissue pattern; it does not make every stained compartment specific.
IF/ICC Q: What should fluorescence show?	A: Membrane associated signal is plausible and a secreted fraction is possible (UniProt Q14114 localisation; HPA: subcellular summary). HPA supplies no ICC/IF images or main cellular location here (HPA: subcellular record), so this section cannot establish an observed fluorescent pattern.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Testis positive control is blank.	The positive cell population may be absent from the examined area, or the staining run may have failed (HPA: High in pachytene spermatocytes; general IHC practice).	Locate pachytene spermatocytes on the section, inspect the control run, and review the IHC validated antibody's supplied procedure before interpreting experimental negatives (general IHC practice).
Strong signal is confined to nuclei.	Nuclear off target binding was observed and disregarded by HPA (HPA: reliability description). A nuclear only result is therefore insufficient evidence of LRP8.	Compare nuclear signal with cell associated staining in the positive population and inspect a no primary control for detection background (HPA: testis IHC; general IHC practice).
Negative reference cells also stain.	HPA found no detection in adipocytes and adrenal glandular cells (HPA: tissue IHC). Unexpected staining could arise from cross reactivity or endogenous chromogenic detection activity (general IHC practice).	Verify the scored cell type; compare matched negative reference and no primary sections, and check the routine endogenous enzyme blocking step for the chosen detection chemistry (general IHC practice).
The entire section has diffuse brown background.	Nonspecific binding, incomplete washing, or chromogen deposition can obscure cellular staining (general IHC practice). UniProt documented shedding alone does not identify the haze as LRP8 (UniProt Q14114 processing).	Check no primary control, blocking, washing, and chromogen development against the run's routine controls; score LRP8 only where cellular localisation remains interpretable (general IHC practice).
Signal appears extracellular or away from cell borders.	Some LRP8 isoforms can yield a secreted receptor fragment (UniProt Q14114 processing), but the antibody epitope and the identity of this deposited signal are unknown.	Record extracellular signal separately from cell associated staining; use the testis positive population and matched controls to judge specificity without assigning an isoform (HPA: testis IHC; general IHC practice).
Results differ between tissue sections despite similar processing.	The compared fields may contain different cell populations; HPA's positive and negative calls are cell specific, and its secreted protein profile complicates tissue level comparisons (HPA: tissue IHC and reliability description).	Compare equivalent cell populations and compartments, document section morphology, and avoid treating an unlisted cell type as a validated positive or negative (general IHC practice).

Source: UniProt Q14114 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing LRP8 staining

IHC analysis of ApoER2/LRP8 using anti-ApoER2/LRP8 antibody (A03444-2). ApoER2/LRP8 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ApoER2/LRP8 Antibody (A03444-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

A03444-2 · IHCIHC analysis of ApoER2/LRP8 using anti-ApoER2/LRP8 antibody (A03444-2). ApoER2/LRP8 was detected in paraffin-embedded section of human lung

IF analysis of ApoER2/LRP8 using anti-ApoER2/LRP8 antibody (A03444-2). ApoER2/LRP8 was detected in immunocytochemical section of HepG2 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 4μg/mL rabbit anti-ApoER2/LRP8 Antibody (A03444-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.

A03444-2 · IFIF analysis of ApoER2/LRP8 using anti-ApoER2/LRP8 antibody (A03444-2). ApoER2/LRP8 was detected in immunocytochemical section of HepG2 cells

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000157193-LRP8/tissue/cerebral+cortex#imid_20366453

HPA · Thyroid glandHuman Protein Atlas IHC, Thyroid gland. Source: https://www.proteinatlas.org/ENSG00000157193-LRP8/tissue/thyroid+gland#imid_20366405

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000157193-LRP8/tissue/colon#imid_20366363

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000157193-LRP8/tissue/liver#imid_20366372

← swipe / scroll for more images →

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

Sample controls for LRP8 IHC & IF

🧪Run testis first and look for staining in pachytene spermatocytes (HPA: High in pachytene spermatocytes). Run adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the testis slide, compare staining with neighboring non-pachytene cells, whose LRP8-negative status is not established by the supplied HPA row.

Positive control tissue: Testis (Pachytene spermatocytes, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for LRP8; derive a cell-line control from the positive tissue's cell type (Pachytene spermatocytes) and confirm it by RNA or western blot first.

Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody), and LRP8 knockout tissue or a validated immunizing-peptide block where available. Check testis sections for endogenous peroxidase and biotin background before interpreting DAB staining, especially with the caption’s biotin-based detection (selected-SKU caption: SABC with DAB).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected-SKU paraffin-section caption is unreported (selected-SKU caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used in that IHC example, but a retrieval requirement has not been established (selected-SKU caption: heat-mediated EDTA retrieval). The supplied evidence does not show that frozen sections or IF are easier; HPA lists no ICC-IF image cell lines, and testis background from the biotin-based detection system should be checked (HPA subcellular: no ICC-IF image cell lines; selected-SKU caption: SABC with DAB).

HPA tissue IHC evidence for LRP8

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex. Nuclear 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.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Testis	Pachytene spermatocytes	High	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 →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →

Section 3

Advanced LRP8 IHC Tips

Use compartment-aware controls and the documented paraffin-section workflow to assess LRP8 staining in chromogenic IHC.

Which retrieval conditions should I try when LRP8 staining is weak in paraffin sections?

Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A03444-2). The documented paraffin-section example used that retrieval before 1 μg/mL primary antibody overnight at 4°C (caption A03444-2). If signal remains weak, check section adhesion, heating consistency, antibody incubation and a known positive control before changing retrieval (standard IHC practice). You may then compare a shorter or longer heating period, keeping detection and exposure conditions constant; the payload supplies no validated alternative buffer or heating time for this antibody (caption A03444-2). Score membrane-associated and plausible extracellular staining separately, because processed LRP8 can yield a secreted fragment (UniProt Q14114 processing).

Could fixation explain variable LRP8 staining across paraffin sections?

Target-specific fixation sensitivity is unknown: the catalog image describes a paraffin-embedded section but does not state its fixative (caption A03444-2). Record the fixative, time before fixation, fixation duration and section age for each specimen so differences can be investigated systematically (standard IHC practice). Compare similarly processed positive-control sections in the same staining run, using the documented EDTA pH 8.0 retrieval and 1 μg/mL primary condition as a starting point (caption A03444-2). If signal varies, optimise processing on matched material before attributing the change to LRP8 biology (standard IHC practice). Neither tissue staining patterns nor receptor topology establish how this particular epitope responds to fixation (HPA tissue IHC; UniProt Q14114 topology).

How should I assess membrane, cytoplasmic and extracellular LRP8 staining?

Assess cell-border staining first: LRP8 has an extracellular region at residues 42–826, a membrane span at 827–847 and a cytoplasmic tail at 848–963 (UniProt Q14114 topology). Cytoplasmic signal can be considered alongside its membrane-receptor and possible endocytic roles, but the antibody's epitope and staining specificity need independent validation (UniProt Q14114 function; standard IHC practice). Record extracellular staining separately because isoforms containing a furin-type cleavage site can produce a secreted receptor fragment (UniProt Q14114 processing). Do not count diffuse extracellular deposit as membrane-positive cells when reporting cellular scores (standard IHC practice). Treat isolated nuclear staining cautiously because nuclear off-target binding was observed and disregarded in the HPA assessment (HPA tissue IHC).

Can this antibody distinguish membrane LRP8 from its processed isoforms?

Do not assume isoform discrimination without the antibody's mapped epitope: LRP8 has 5 listed isoforms (UniProt Q14114 isoforms). An epitope in residues 42–826 could occur on the extracellular receptor region, while one in residues 848–963 lies on the cytoplasmic tail (UniProt Q14114 topology). Isoforms containing the furin-type cleavage-site exon can release a secreted receptor fragment, so extracellular signal needs its own interpretation (UniProt Q14114 processing). Extracellular-region glycosylation sites include residues 176, 441, 518, 538, 772 and 807; their presence does not establish antibody sensitivity to processing (UniProt Q14114 glycosylation). Obtain epitope documentation or validate with orthogonal controls before assigning staining to a specific isoform (standard IHC practice).

How can I investigate LRP8 localisation with multiplex IF?

Use this as an exploratory IF comparison with the chromogenic result; the supplied catalog example documents paraffin-section IHC, without an IF staining condition (caption A03444-2). Pair LRP8 with a validated marker for pachytene spermatocytes when examining testis, since that cell population is reported as strongly positive (HPA tissue IHC). Choose a far-red LRP8 fluorophore where tissue autofluorescence affects shorter wavelengths, and check single-stain and no-primary controls before interpreting colocalisation (standard IF practice). For an extracellular-domain epitope, test minimal permeabilisation first; for a cytoplasmic-tail epitope, test controlled permeabilisation, because the domains occupy opposite sides of residues 827–847 (UniProt Q14114 topology; standard IF practice).

What should I check when LRP8 DAB staining appears widespread?

Run a no-primary control and inspect endogenous pigment, tissue edges and necrotic areas before treating widespread brown signal as LRP8 (standard IHC practice). The documented workflow used 10% goat serum, 1 μg/mL primary overnight at 4°C, a biotinylated secondary and streptavidin–biotin DAB detection (caption A03444-2). Include a peroxidase block and appropriate secondary-only controls as general chromogenic IHC precautions, particularly when background follows the detection system (standard IHC practice). Titrate primary antibody and DAB development while keeping retrieval at EDTA pH 8.0 during initial troubleshooting (caption A03444-2; standard IHC practice). Disregard isolated nuclear signal unless independently validated, given the reported nuclear off-target binding (HPA tissue IHC).

How should I score LRP8 IHC across specimens with different cell compositions? ⚠ ANSWER MARKED FOR VERIFICATION

Define the scored cell population and compartment before analysis; report membrane-associated, cytoplasmic and extracellular signal separately (UniProt Q14114 topology and processing; standard IHC practice). For cellular staining, record the percentage of positive cells and intensity from 0–3, then calculate an H-score from 0–300 if that scale suits the study (standard IHC practice). Normalise positive-cell counts to the number of eligible cells, or stained area to tissue area in mm², rather than comparing raw fields (standard IHC practice). Use the same section selection, retrieval and DAB development criteria across specimens, and exclude folds, necrosis and edge staining consistently (standard IHC practice). Report the fraction of pachytene spermatocytes separately when using testis controls (HPA tissue IHC).

What evidence separates true LRP8 staining from artefact?

A convincing result should recur in the expected cell population and plausible compartment, with appropriate staining controls and consistent processing (standard IHC practice; UniProt Q14114 topology). Pachytene spermatocytes are a reported strong testis reference, whereas adipocytes and several listed glandular or epithelial populations were not detected in the HPA survey (HPA tissue IHC). Treat nuclear-only staining as suspect because HPA observed nuclear off-target binding and disregarded it (HPA tissue IHC). Investigate edge accentuation, necrosis and residual endogenous peroxidase with morphology and no-primary controls before calling DAB signal positive (standard IHC practice). Extracellular signal may represent a processed receptor fragment, so do not equate it automatically with intact surface receptor (UniProt Q14114 processing).

Boster reagents

Best LRP8 / Low-density lipoprotein receptor-related protein 8 IHC Antibodies

A03444-2 has IHC images from human paraffin-embedded lung, mammary and rectal cancer sections, plus an IF image from HepG2 cells (catalog image captions). Human, mouse and rat reactivity is listed (catalog).

Real IHC data

Anti-ApoER2/LRP8 Antibody ®

Cat # A03444-2

View product →

A03444-2 will render with its human lung cancer paraffin-section IHC figure; additional IHC captions show human mammary and rectal cancer sections (catalog IHC image captions). Its IF caption shows HepG2 cells, while the application list includes IHC, ICC and IF and lists human, mouse and rat reactivity (catalog IF image caption; catalog applications and reactivity).

Which to pick: Choose A03444-2 for paraffin-section IHC because its own IHC caption documents EDTA retrieval at pH 8.0 and primary antibody at 1 μg/ml (catalog IHC image caption); the fixative is unreported (catalog IHC image caption). For IF/ICC, the same SKU has a HepG2 IF image and a listed IF/ICC concentration of 4 μg/ml (catalog IF image caption; datasheet). For mouse or rat samples, the same SKU lists reactivity, though the supplied IHC and IF images document human samples (catalog reactivity; catalog image captions).

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

References

UniProt Consortium. UniProt entry Q14114 (LRP8_HUMAN, Low-density lipoprotein receptor-related protein 8).
Human Protein Atlas. LRP8 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. LRP8 subcellular location (ICC-IF): Membrane, Secreted.
Human Protein Atlas. LRP8 antibody validation summary (1 antibodies).
Reelin-LRP8 signaling mediates brain dissemination of breast cancer cells via abluminal migration. EMBO molecular medicine 2025 — PMC12339728.
Enhanced LRP8 expression induced by Helicobacter pylori drives gastric cancer progression by facilitating β-Catenin nuclear translocation. Journal of advanced research 2025 — PMC11954824.
LRP8 Promotes colorectal cancer progression by suppressing ferroptosis through the SLC3A2/GPX4 signalling axis. European journal of medical research 2026 — PMC12964802.
LRP8 Regulates Lipid Metabolism to Stimulate Malignant Progression and Cisplatin Resistance in Bladder Cancer. The Kaohsiung journal of medical sciences 2025 — PMC12412626.
PubMed PMID:8626535 — UniProt-cited evidence.
PubMed PMID:9079678 — UniProt-cited evidence.
PubMed PMID:11152697 — UniProt-cited evidence.