ELOVL6 / Very long chain fatty acid elongase 6 · IHC design guide

Design Immunohistochemistry for ELOVL6

Plan ELOVL6 IHC in paraffin sections using the observed cytoplasmic tissue pattern (HPA tissue IHC). Compare stained cells with appropriate controls because the tissue staining has uncertain reliability (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for ELOVL6 (IHC for ELOVL6): expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A06004-1, validated IHC image, and IHC protocol steps
Printable ELOVL6 IHC protocol sheet — expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A06004-1, controls and protocol steps. Open the full ELOVL6 IHC guide →

ELOVL6 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 Cytoplasmic staining in several tissues (HPA tissue IHC)
Staining pattern Cytoplasmic staining in adipocytes and glandular cells (HPA tissue IHC)
Antigen retrieval Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen)
Positive control ⓘ Adipose tissue+4 more · see all
Negative control ⓘ Caudate+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). Selected-image fixative and duration unreported (datasheet A06004-1); verify before use.
Caveat Tissue staining has uncertain reliability (HPA tissue IHC)
Regulation No expression regulator reported (UniProt)
Isoform / epitope No isoforms; 1–265 chain, epitope side unassigned (UniProt)
Section 1

Recommended ELOVL6 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is followed by published ELOVL6 IHC methods for normal, polyp, and COAD tissues (PMC9755665) and paraffin-embedded patient tissues (PMC9937776).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleTissue sections; selected-image fixative not specified (standard IHC workflow)
FixationImage fixative and duration unreported (datasheet A06004-1); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibodyRabbit anti-ELOVL6, 5 μg/mL (datasheet A06004-1)
Primary incubationOvernight at 4 °C (standard)
DetectionHRP-polymer secondary, DAB chromogen 5–10 min (standard)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultELOVL6-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control.
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); ELOVL6 is an ER membrane protein (UniProt Q9H5J4).
Section 2

What Is the Expected ELOVL6 Staining Pattern?

ELOVL6 is a 7-pass endoplasmic reticulum membrane protein (UniProt Q9H5J4 topology and subcellular location). In paraffin-section IHC, expect cytoplasmic staining in selected cell populations, including adipocytes and glandular cells in several tissues (HPA tissue IHC: Medium). Interpret this as a provisional pattern: HPA rates tissue staining reliability Uncertain, with only medium agreement between antibody staining and RNA expression (HPA tissue IHC reliability).

What am I looking at on my slide?
Cytoplasmic chromogen in adipocytes or glandular cells, with little dominant nuclear staining (HPA tissue IHC: Medium in selected populations).This fits the reported cytoplasmic profile and the ER membrane location (HPA tissue IHC profile; UniProt Q9H5J4 subcellular location). A fine granular or reticular appearance is plausible for an ER protein, but ordinary chromogenic IHC cannot establish precise organelle colocalisation (standard IHC interpretation). Judge the cell population as well as the compartment.
Predominantly nuclear, extracellular, or sharply cell-surface staining instead of cytoplasmic signal (HPA tissue IHC profile).This conflicts with the reported cytoplasmic staining and ER membrane assignment (HPA tissue IHC profile; UniProt Q9H5J4 subcellular location). Review tissue morphology, chromogen deposits, and controls before assigning ELOVL6 identity (standard IHC practice). A surface-like edge alone does not prove membrane localisation at the organelle level (standard IHC interpretation).
Strong staining in a population reported as not detected, such as esophageal squamous epithelium or splenic red-pulp cells (HPA tissue IHC).Treat unexpected signal as a specificity question, including possible cross-reactivity or endogenous detection activity (standard IHC practice). HPA's 'Not detected' calls describe those sampled cell populations; they do not establish an absolute biological absence (HPA tissue IHC). Compare a reported Medium population and appropriate detection controls on the same run (HPA tissue IHC; standard IHC practice).
Uniform haze across tissue, stroma, and empty spaces, obscuring cell boundaries (standard IHC interpretation).This is a background pattern, not the cell-restricted cytoplasmic profile reported for ELOVL6 (HPA tissue IHC profile). Nonspecific reagent binding, endogenous enzyme activity, or excessive chromogen development can produce broad staining (standard chromogenic IHC practice). Assess the no-primary control and slide morphology before scoring intensity (standard IHC practice).
No convincing cytoplasmic signal in adipocytes despite an intact section (HPA tissue IHC: Medium in adipocytes).A failed positive reference is a run-level warning, although HPA's Uncertain reliability prevents treating any one specimen as guaranteed positive (HPA tissue IHC reliability and adipose-tissue result). Check control-slide performance, reagent delivery, retrieval conditions, and detection before calling the study tissue negative (standard IHC practice).
💡Expected ELOVL6 appearanceCall a result provisionally positive when Medium cytoplasmic staining is resolved in a reported population such as adipocytes or selected glandular cells (HPA tissue IHC); dominant nuclear signal, tissue-wide haze, or strong signal in a reported Not detected population warrants specificity checks (UniProt Q9H5J4 subcellular location; HPA tissue IHC; standard IHC interpretation).
How each factor affects the staining
Membrane topology and visual resolution (UniProt Q9H5J4 topology).ELOVL6 has 7 transmembrane segments and an ER membrane assignment (UniProt Q9H5J4). Cytoplasmic chromogen is compatible with that location, but light-microscope IHC cannot demonstrate that the colored material lies in the ER membrane (standard IHC interpretation).
Cell-population differences (HPA tissue IHC).HPA reports Medium staining in adipocytes, several glandular populations, Purkinje cells, and colon endothelial cells; it reports Low or Not detected staining in other listed populations (HPA tissue IHC). UniProt's 'ubiquitous' tissue-specificity annotation does not imply equal detectable IHC signal in every cell (UniProt Q9H5J4 tissue specificity; HPA tissue IHC).
Evidence strength for the staining pattern (HPA tissue IHC; HPA antibody HPA042355).HPA calls the tissue pattern Uncertain, notes medium staining–RNA consistency, and lists HPA042355 IHC validation as Uncertain (HPA tissue IHC reliability; HPA antibody HPA042355). Use reported positive and negative populations as comparisons, with controls, rather than as definitive proof of antibody specificity (standard IHC interpretation).
Processing and fixation evidence (UniProt Q9H5J4 processing; supplied source scope).UniProt annotates a full-length 1–265 chain, with no signal peptide or propeptide and no annotated isoforms (UniProt Q9H5J4 processing and isoforms). The supplied sources do not establish ELOVL6-specific sensitivity to fixation or retrieval; choose and assess those conditions as general IHC workflow variables (standard IHC practice).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
Adipocyte reference is blank, and study sections are also blank (HPA tissue IHC: adipocytes Medium).The run may have failed at staining, retrieval, reagent delivery, or detection; the reference itself is not guaranteed positive in every specimen (standard IHC practice; HPA tissue IHC reliability: Uncertain).Inspect section integrity and a positive-reference slide; verify the documented IHC-P procedure and reagent steps, then repeat with an appropriate detection control before interpreting biological absence (standard IHC practice).
Expected cytoplasmic signal is weak beside heavy counterstain (HPA tissue IHC profile).Counterstain or chromogen development can make a weak cytoplasmic reaction hard to resolve (standard chromogenic IHC practice). HPA reports Low staining in some populations, including lung macrophages and cardiomyocytes (HPA tissue IHC).Compare the same cell type across control and study slides; adjust counterstain or development within the validated IHC workflow, then score only clearly resolved cellular signal (standard IHC practice).
Nuclei dominate the stain in an otherwise interpretable section (HPA tissue IHC profile).That compartment disagrees with cytoplasmic HPA staining and UniProt's ER membrane assignment; nonspecific staining or a slide artifact is possible (HPA tissue IHC; UniProt Q9H5J4; standard IHC interpretation).Check the no-primary control, chromogen deposits, and cell boundaries. Reassess antibody specificity before recording nuclear ELOVL6 localisation (standard IHC practice).
A reported Not detected population stains strongly (HPA tissue IHC).Cross-reactivity or endogenous detection activity is possible, while biological variation cannot be excluded from HPA's limited, Uncertain IHC evidence (standard IHC practice; HPA tissue IHC reliability).Compare it with a reported Medium population and no-primary control in the same run. Record the exact cell type and compartment rather than treating whole-tissue staining as one result (HPA tissue IHC; standard IHC practice).
Diffuse brown haze makes nearly every cell look positive (standard IHC interpretation).Nonspecific binding, endogenous enzyme activity, or overdevelopment may obscure the cell-restricted pattern (standard chromogenic IHC practice; HPA tissue IHC profile).Review background in the no-primary control, blocking and wash steps, and chromogen development; repeat scoring only when cellular staining separates from background (standard IHC practice).
Can IF/ICC confirm the precise ELOVL6 compartment? (HPA subcellular ICC-IF).HPA gives only a broad 'Membrane' summary, no main location, and no cell-line ICC-IF images; it cannot verify an ER pattern by imaging here (HPA subcellular ICC-IF).Use UniProt's ER membrane assignment as the localisation hypothesis and consult the separate IF/ICC guide for that application; do not infer an IF protocol or validated colocalisation from this IHC evidence (UniProt Q9H5J4; HPA subcellular ICC-IF).

Sample controls for ELOVL6 IHC & IF

🧪Run adipose tissue first and look for staining in adipocytes (HPA: Medium in adipocytes); use esophageal squamous epithelium as a negative comparison (HPA: Not detected in esophageal squamous epithelial cells). On the positive slide, nonadipocyte cells should show only background in the no-primary control; do not assume they are biologically ELOVL6-negative (UniProt: ubiquitous tissue specificity).
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ELOVL6; derive a cell-line control from the positive tissue's cell type (Adipocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and an irrelevant IgG control matched to the primary antibody’s host species and class (standard IHC practice); use validated ELOVL6 knockout material as a biological negative if available (standard IHC practice). Quench endogenous peroxidase and inspect background in the adipose section before scoring its thin adipocyte cytoplasmic rims (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval effect are unreported in the supplied evidence; optimize antigen retrieval empirically for this ER membrane protein (UniProt: endoplasmic reticulum membrane). The selected tissue-IHC caption reports human brain staining at 5 µg/mL, but its fixative is unreported (catalog caption: A06004-1). For IF/ICC, the supplied evidence does not establish that frozen sections or IF are easier; assess signal and autofluorescence separately, especially around adipocyte lipid vacuoles (standard IF practice).

HPA tissue IHC evidence for ELOVL6

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes Medium Protein (IHC) HPA →
Adrenal gland Glandular cells Medium Protein (IHC) HPA →
Appendix Glandular cells Medium Protein (IHC) HPA →
Cerebellum Purkinje cells Medium Protein (IHC) HPA →
Colon Endothelial cells Medium Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Caudate Glial cells Not detected Protein (IHC) HPA →
Cervix Glandular cells Not detected Protein (IHC) HPA →
Endometrium Cells in endometrial stroma Not detected Protein (IHC) HPA →
Esophagus Squamous epithelial cells Not detected Protein (IHC) HPA →
Hippocampus Glial cells Not detected Protein (IHC) HPA →
Section 3

Advanced ELOVL6 IHC Tips

Troubleshoot ELOVL6 chromogenic IHC in paraffin sections by checking retrieval, compartmental staining, controls and scoring (UniProt Q9H5J4; HPA tissue IHC).

What retrieval should I start with for weak ELOVL6 staining?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 minutes (page retrieval rule: cytoplasmic / membrane antigen). Keep section thickness, heating vessel and cooling time consistent while comparing runs (standard IHC practice). ELOVL6 has 7 transmembrane segments, so epitope accessibility may depend on the antibody and retrieval conditions (UniProt Q9H5J4 topology; standard IHC practice). If staining remains weak, test an alternative retrieval buffer on matched sections as a fallback, alongside the citrate condition (standard IHC practice). Compare signal in expected cells with background and a no-primary control before adopting any change (HPA tissue IHC; standard IHC practice).
Could fixation explain variable ELOVL6 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown from the supplied evidence; the selected brain IHC caption does not state a fixative (catalog antibody A06004-1 caption). Record each block’s fixative and fixation duration, then compare sections processed with the same retrieval and detection settings (standard IHC practice). Prolonged or inconsistent fixation can alter antigen accessibility in paraffin IHC generally, but it does not establish a particular effect on ELOVL6 (standard IHC practice). Use a consistently processed reference section and a no-primary control in each comparison (standard IHC practice). Interpret a difference between blocks cautiously because HPA staining reliability is listed as uncertain (HPA tissue IHC).
Where should ELOVL6 staining appear in tissue sections?
Expect predominantly cytoplasmic, membrane-associated staining rather than a nuclear pattern: ELOVL6 is an endoplasmic reticulum membrane protein (UniProt Q9H5J4 subcellular location). At chromogenic IHC resolution, an ER-associated signal may appear finely granular or diffuse within the cytoplasm, without resolving individual membranes (UniProt Q9H5J4 subcellular location; standard IHC practice). HPA describes cytoplasmic expression across several tissues, but rates its tissue staining reliability as uncertain (HPA tissue IHC). Assess the signal within identified cells and compare it with a no-primary control before assigning localisation (standard IHC practice). Strong isolated nuclear or extracellular staining warrants investigation as possible artefact (UniProt Q9H5J4 subcellular location; standard IHC practice).
How should I troubleshoot an antibody whose ELOVL6 epitope is unspecified?
The supplied record annotates no ELOVL6 isoforms and describes a 265-amino-acid chain; this does not identify the catalog antibody’s epitope (UniProt Q9H5J4 processing and isoforms; catalog antibody payload). ELOVL6 has 7 transmembrane segments and one annotated glycosylation site at residue 2 (UniProt Q9H5J4 topology and glycosylation). Without an epitope sequence and membrane orientation for that epitope, do not assume which side retrieval must expose (UniProt Q9H5J4 topology; standard IHC practice). Ask for epitope information when available, and compare retrieval conditions on adjacent sections using the same detection settings (standard IHC practice). Judge specificity by appropriate cellular staining and controls, not staining intensity alone (HPA tissue IHC; standard IHC practice).
How can I check ELOVL6 localisation with tissue immunofluorescence?
Use IF as a separate optimisation, since the supplied catalog evidence shows tissue chromogenic IHC and provides no IF validation (catalog antibody A06004-1 caption). Multiplex ELOVL6 with an adipocyte marker in adipose tissue to identify a reported medium-staining cell population (HPA tissue IHC; standard IF practice). Choose a fluorophore channel after imaging an unstained section for tissue autofluorescence, and include single-label controls for bleed-through (standard IF practice). Because ELOVL6 spans the ER membrane 7 times, select permeabilisation only after establishing whether the antibody epitope faces the cytosol or ER lumen (UniProt Q9H5J4 topology; standard IF practice). Compare the resulting intracellular pattern with the chromogenic IHC control tissue (standard IF practice).
How do I reduce diffuse or granular ELOVL6 background?
First inspect a no-primary section to separate detection background from antibody-associated staining (standard IHC practice). For chromogenic detection, block endogenous peroxidase before adding the primary antibody, then check secondary-only and substrate controls as appropriate (standard IHC practice). Titrate the catalog antibody and detection reagent on matched sections, keeping retrieval fixed at citrate pH 6.0 while comparing background (page retrieval rule; standard IHC practice). Evaluate granular signal against cell boundaries: ELOVL6 is ER membrane-associated, while HPA reports cytoplasmic tissue staining with uncertain reliability (UniProt Q9H5J4 subcellular location; HPA tissue IHC). Avoid scoring precipitate, folds or section edges as cellular signal (standard IHC practice).
What is a defensible way to score ELOVL6 chromogenic IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because ELOVL6 is ER membrane-associated and HPA reports cytoplasmic staining (UniProt Q9H5J4 subcellular location; HPA tissue IHC). For comparable sections, report the percentage of positive cells and an H-score from 0–300, using intensity categories 0–3 (standard IHC practice). If counting stained cells by area, express density per mm² of viable tissue and state the area excluded (standard IHC practice). Normalise comparisons to the same cell type, section area, staining batch and exposure or scanner settings (standard IHC practice). Include a reference section because HPA rates tissue staining reliability as uncertain (HPA tissue IHC; standard IHC practice).
How can I distinguish genuine ELOVL6 signal from staining artefact?
Prioritise intracellular cytoplasmic staining in intact cells: ELOVL6 resides in the ER membrane, and HPA describes cytoplasmic tissue expression (UniProt Q9H5J4 subcellular location; HPA tissue IHC). HPA reports medium staining in adipocytes and cerebellar Purkinje cells, but its overall tissue IHC reliability is uncertain (HPA tissue IHC). Treat signal confined to nuclei, section edges, folds or necrotic areas as suspect until controls support it (UniProt Q9H5J4 subcellular location; standard IHC practice). Check a no-primary section and endogenous peroxidase block when interpreting brown chromogen deposits (standard IHC practice). Compare candidate positives with expected cell identity and repeatable localisation across matched sections before drawing biological conclusions (HPA tissue IHC; standard IHC practice).
Boster reagents

Best ELOVL6 / Very long chain fatty acid elongase 6 IHC Antibodies

A06004-1 has IHC and IF images from human brain tissue (catalog: image captions) and lists Human, Mouse, and Rat reactivity (catalog: reactivity).

Real IHC data Immunohistochemistry of ELOVL6 in human brain tissue with ELOVL6 antibody at 5 μg/mL.
Anti-ELOVL6 Antibody
Cat # A06004-1

A06004-1 will render with a human brain tissue IHC image at 5 μg/mL (IHC image caption). It also lists IHC-P and IF, with a human brain tissue IF image at 20 μg/mL (catalog: applications; IF image caption).

Which to pick: Choose A06004-1 for paraffin-section IHC: IHC-P is listed, and its own image caption documents human brain tissue staining at 5 μg/mL; the fixative is unreported (catalog: applications; IHC image caption). For IF, its human brain tissue image uses 20 μg/mL; ICC validation and clonality are unreported (IF image caption; catalog: applications, clone). For Mouse or Rat samples, A06004-1 lists reactivity with both species, while the supplied IHC and IF images show Human brain tissue (catalog: reactivity; image captions).

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

References

  1. UniProt Consortium. UniProt entry Q9H5J4 (ELOV6_HUMAN, Very long chain fatty acid elongase 6).
  2. Human Protein Atlas. ELOVL6 tissue IHC expression (reliability: Uncertain).
  3. Human Protein Atlas. ELOVL6 subcellular location (ICC-IF): Membrane.
  4. Human Protein Atlas. ELOVL6 antibody validation summary (1 antibodies).
  5. A novel enterocyte-related 4-gene signature for predicting prognosis in colon adenocarcinoma. Frontiers in immunology 2022 — PMC9755665.
  6. Elovl6 is a negative clinical predictor for liver cancer and knockdown of Elovl6 reduces murine liver cancer progression. Scientific reports 2018 — PMC5920119.
  7. An Investigation of the Prognostic Role of Genes Related to Lipid Metabolism in Head and Neck Squamous Cell Carcinoma. International journal of genomics 2023 — PMC9937776.
  8. Exploring the role of ELOVLs family in lung adenocarcinoma based on bioinformatic analysis and experimental validation. BMC cancer 2025 — PMC11720344.
  9. PubMed PMID:14702039 — UniProt-cited evidence.
  10. PubMed PMID:15815621 — UniProt-cited evidence.
  11. PubMed PMID:15489334 — UniProt-cited evidence.