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KDELR3 / ER lumen protein-retaining receptor 3 · IHC design guide
Design Immunohistochemistry for KDELR3
Plan chromogenic KDELR3 IHC in paraffin sections using glandular cells in appendix and small intestine as positive references (HPA tissue IHC). Expect cytoplasmic, sometimes granular staining, but interpret it cautiously because tissue-IHC reliability is uncertain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KDELR3 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, sometimes granular (HPA tissue IHC) | |
| Staining pattern | Glandular cells: cytoplasmic, sometimes granular (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+4 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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A14682-1) | |
| Caveat | Very low staining–RNA concordance (HPA tissue IHC) | |
| Regulation | No regulation annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; map epitopes to lumenal or cytoplasmic loops (UniProt) |
Section 1
Recommended KDELR3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol appears alongside four published KDELR3 chromogenic IHC examples (PMC11681132; PMC11625429; PMC11451269; PMC6965108).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A14682-1) |
| Fixation | Image fixative and duration unreported (datasheet A14682-1); 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-KDELR3, 1:50-1:200 (datasheet A14682-1) |
| 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 | KDELR3-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic, sometimes granular expression in a few 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 setting); adjust dilution using the relevant published tissue protocol.
Section 2
What Is the Expected KDELR3 Staining Pattern?
KDELR3 is a seven-pass membrane receptor that cycles between the Golgi and endoplasmic reticulum with KDEL-bearing cargo (UniProt O43731 topology and localization). In paraffin-section IHC, expect cytoplasmic, sometimes granular staining, particularly in appendix and small-intestine glandular cells (HPA: High). Treat this as a provisional pattern: HPA rates tissue IHC reliability Uncertain because antibody staining and RNA expression show very low consistency (HPA: tissue IHC).
| Glandular cells show cytoplasmic, sometimes granular chromogen in appendix or small intestine (HPA: High in both). | This matches the reported tissue pattern and the ER/Golgi membrane location (HPA: tissue IHC; UniProt O43731 localization). Evaluate cell boundaries and morphology before scoring; HPA's Uncertain IHC rating prevents treating matching stain alone as proof of specificity (HPA: tissue IHC). |
| Most signal appears inside nuclei, along cell surfaces, or in extracellular material. | Those dominant compartments do not match the reported cytoplasmic IHC pattern or membrane-bound ER/Golgi localization (HPA: tissue IHC; UniProt O43731 localization). Review morphology and controls for artefact before interpreting the signal as KDELR3. |
| Unexpected strong stain appears in adipocytes or bronchial respiratory epithelium (HPA: Not detected in those cells). | Investigate antibody cross-reactivity or endogenous detection activity, especially if the stain is diffuse or lacks cytoplasmic structure. A mismatch is a warning, not a definitive exclusion: the HPA tissue IHC assessment is Uncertain (HPA: tissue IHC). |
| Chromogen covers many cell types and the surrounding section without clear cell-associated granules. | Poorly localized, widespread color is more consistent with background than the reported cytoplasmic, sometimes granular pattern (HPA: tissue IHC). Compare a no-primary control and assess blocking, washing, and chromogen development as general IHC checks. |
| Appendix or small-intestine glandular cells have no discernible stain despite preserved morphology (HPA: High in both). | Check that the assay can reveal signal before calling the sample KDELR3-negative. Review antibody identity, retrieval, detection, and the positive-control section as general IHC checks; HPA's Uncertain rating means even these reported high-staining tissues are imperfect controls (HPA: tissue IHC). |
💡Expected KDELR3 appearanceCall a provisional positive when glandular cells show distinct cytoplasmic, sometimes granular staining, strongest in HPA-reported High appendix or small-intestine examples; dominant nuclear, extracellular, or uniform section-wide color is suspect (HPA: tissue IHC; UniProt O43731 localization).
| Cell compartment and topology (UniProt O43731) | KDELR3 has seven transmembrane segments and is assigned to ER, Golgi, and COPI-vesicle membranes (UniProt O43731 topology and localization). Cytoplasmic organelle-associated staining is plausible; topology alone does not specify how a particular antibody will stain a paraffin section. |
| Tissue-control choice (HPA: tissue IHC) | Appendix and small-intestine glandular cells are reported High; adipocytes and bronchial respiratory epithelial cells are Not detected (HPA: tissue IHC). Use these as comparison tissues or cell types while retaining the Uncertain IHC reliability caveat. |
| Antibody evidence (HPA: antibody validation) | HPA043477 has Uncertain IHC status, while HPA076405 has Supported ICC status and no listed IHC status (HPA: antibody validation). The ICC result does not validate either antibody's paraffin-section IHC pattern; keep application-specific evidence separate. |
| Isoforms and epitope (UniProt O43731) | Two isoforms are listed, and their coverage by any particular antibody cannot be inferred from this record (UniProt O43731 isoforms). If staining differs between samples, check the antibody's stated epitope and isoform coverage before assigning the difference to KDELR3 abundance. |
| IF/ICC Q&A: What pattern is supported? | An ER pattern is supported in ICC-IF, with images listed for MCF-7, SiHa, and U2OS (HPA: subcellular). That finding can inform localization interpretation; it does not establish an IF protocol or override the Uncertain tissue IHC assessment (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in appendix or small-intestine glandular cells (HPA: High). | The IHC assay may have failed, or the antibody may not reproduce the reported tissue pattern; HPA rates tissue IHC Uncertain (HPA: tissue IHC). | Confirm the expected cells are present, run a documented positive control, and check retrieval and detection against the antibody's IHC instructions. Treat retrieval review as general IHC practice, not a KDELR3-specific fixation claim. |
| Strong nuclear signal dominates the section. | Nuclear dominance conflicts with the cytoplasmic HPA pattern and ER/Golgi membrane assignment (HPA: tissue IHC; UniProt O43731 localization). | Compare the no-primary control and inspect staining at the cell level. Reassess antibody specificity and detection background before scoring nuclear color as KDELR3. |
| Adipocytes or bronchial respiratory epithelial cells stain strongly (HPA: Not detected). | The observed cell-type pattern may reflect nonspecific binding or endogenous detection activity; the HPA IHC profile is itself Uncertain (HPA: tissue IHC). | Include a no-primary control, review detection blocking, and compare the same run with appendix or small intestine. Investigate the mismatch without declaring the unexpected cells definitively KDELR3-negative. |
| Diffuse chromogen obscures glandular-cell boundaries. | Background from binding, inadequate washing, or detection development can mask the reported cytoplasmic, sometimes granular pattern (HPA: tissue IHC; general IHC practice). | Inspect the no-primary control; optimize blocking, washes, and development using standard IHC practice. Score only distinguishable cell-associated staining, and record any unresolved background. |
| Only a faint or patchy signal appears in a reported High tissue (HPA: appendix and small intestine). | Variation in assay performance or section quality may weaken visible staining; HPA's Uncertain rating also limits how firmly its High calls predict another run (HPA: tissue IHC). | Check preserved tissue morphology, reagent performance, and staining consistency across the section. Repeat with an appropriate control if needed; avoid converting weak signal into a categorical absence call. |
| IF/ICC shows an ER pattern while tissue IHC is weak or inconsistent (HPA: subcellular; HPA: tissue IHC). | The assays have different validation evidence: HPA reports Supported ICC localization for HPA076405 but Uncertain tissue IHC for HPA043477 (HPA: antibody validation). | Interpret each application with its own antibody and controls. Use the ER result as localization context, then evaluate paraffin-section IHC on its own cytoplasmic pattern and tissue controls. |
Experiment evidence
IHC & IF images showing KDELR3 staining
A14682-1 · IHCImmunohistochemistry (IHC) analyzes of KDEL Receptor 3 (L95) pAb in paraffin-embedded human breast carcinoma tissue at 1:50.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100196-KDELR3/tissue/cerebral+cortex#imid_11905819
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100196-KDELR3/tissue/colon#imid_11905745
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100196-KDELR3/tissue/liver#imid_11905754
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000100196-KDELR3/tissue/kidney#imid_11905755
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA076405; green = KDELR3, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA076405; green = KDELR3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KDELR3 IHC & IF
🧪Run appendix first and look for staining in glandular cells (High; HPA: appendix). Use adipose tissue as the negative tissue and assess adipocytes (HPA: Not detected in adipocytes); on the appendix slide, cells without specific staining should show only background, but HPA does not identify a confirmed internal negative cell type there.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KDELR3 in MCF-7, SiHa, U2OS, with annotated localisation: Endoplasmic reticulum (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched isotype control matched to the primary antibody’s host species and clonality (standard IHC practice). Use KDELR3 knockout tissue or a validated immunizing-peptide block as a biological specificity control; for chromogenic appendix IHC, quench endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval requirement are unreported in the supplied evidence; evaluate antigen retrieval empirically for paraffin sections. The selected catalog antibody’s breast carcinoma IHC caption reports paraffin sections at 1:50, but its fixative is unreported (caption: A14682-1). The evidence does not establish that frozen sections or IF are easier; in appendix, distinguish glandular-cell staining from endogenous chromogenic background (HPA: High in appendix glandular cells; standard IHC practice).
HPA tissue IHC evidence for KDELR3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Very low 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KDELR3 IHC Tips
Troubleshoot KDELR3 staining in paraffin sections by checking retrieval, controls, subcellular pattern, and scoring before interpreting tissue differences.
What retrieval conditions should I start with for KDELR3 in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval specification). After cooling and washing, compare a small retrieval time series on matched sections while keeping antibody concentration and chromogen development constant (standard IHC practice). KDELR3 has 7 transmembrane segments with short lumenal and cytoplasmic regions, so epitope accessibility may depend on the antibody's binding site, which is not supplied here (UniProt O43731 topology; antibody information supplied). If signal remains weak, test another retrieval buffer as a fallback alongside the specified citrate condition, and judge improvement against background and compartmental pattern (standard IHC practice; UniProt O43731 localisation).
Could fixation explain weak KDELR3 staining in my paraffin sections?
KDELR3-specific fixation sensitivity is unknown: the selected paraffin-section caption reports a 1:50 antibody dilution but does not state the fixative (A14682-1 tissue-IHC caption). Record the fixative and fixation duration for each specimen, then compare sections processed together before assigning a weak signal to the target (standard IHC practice). If fixation histories differ, use matched control tissue and the same retrieval, antibody incubation, and chromogen development to assess whether the staining difference persists (standard IHC practice). Do not derive a KDELR3 fixation requirement from its membrane topology or reported tissue pattern; neither establishes fixation sensitivity (UniProt O43731 topology; HPA tissue IHC).
Where should convincing KDELR3 staining appear in a tissue section?
Expect predominantly cytoplasmic staining with a granular or perinuclear distribution consistent with the endoplasmic reticulum and Golgi, rather than a nuclear-only pattern (HPA tissue IHC; UniProt O43731 localisation). KDELR3 resides in endoplasmic-reticulum, Golgi, and COPI-coated vesicle membranes and cycles with KDEL-bearing cargo, so a single static compartment need not capture its full distribution (UniProt O43731 localisation and function). Assess the pattern within intact cells using the counterstain to identify nuclei and tissue boundaries (standard IHC practice). Investigate diffuse extracellular deposits or isolated nuclear signal with a no-primary control and a repeat section before calling them KDELR3-positive (standard IHC practice; UniProt O43731 localisation).
Can this antibody distinguish KDELR3 isoforms after antigen retrieval?
KDELR3 has 2 listed isoforms, but the supplied antibody information does not identify its epitope or establish isoform selectivity (UniProt O43731 isoforms; A14682-1 tissue-IHC caption). Its 7 transmembrane segments leave short lumenal and cytoplasmic regions, making the epitope's position relevant to accessibility in sections (UniProt O43731 topology). Check the antibody's documented immunogen or epitope against isoform sequences before claiming isoform-specific staining, and retain the same retrieval condition when comparing sections (standard IHC practice; page retrieval specification). If the epitope remains unknown, report staining as KDELR3 immunoreactivity and support any isoform assignment with an independent isoform-resolving method (standard IHC practice; UniProt O43731 isoforms).
How should I compare KDELR3 immunofluorescence with chromogenic IHC?
Treat IF as a separate validation context: HPA reports endoplasmic-reticulum localisation in ICC/IF, while the selected product caption documents paraffin-section IHC at 1:50 without stating a fixative (HPA subcellular; A14682-1 tissue-IHC caption). For IF multiplexing, pair KDELR3 with a marker identifying the expected cell population and choose a fluorophore channel with low tissue autofluorescence after inspecting an unstained control (standard IF practice). If the antibody epitope is cytoplasmic, permeabilisation must provide cytoplasmic access; a lumenal epitope may require access across organelle membranes, so optimise conditions after establishing the epitope side (UniProt O43731 topology; standard IF practice). Compare compartmental patterns across methods without treating fluorescence intensity as equivalent to chromogenic staining intensity (standard IHC/IF practice).
How can I reduce diffuse or granular background without losing KDELR3 signal?
First compare the stained section with a no-primary control and inspect whether granules follow intact cell boundaries or occur across empty tissue spaces (standard IHC practice). Optimise blocking, primary-antibody concentration, wash conditions, and chromogen development one variable at a time; the selected image uses 1:50, which documents that image rather than a universal working dilution (standard IHC practice; A14682-1 tissue-IHC caption). Include a peroxidase block when using peroxidase-based chromogenic detection, and examine pigment or endogenous enzyme signal in control sections (standard IHC practice). Retain staining that reproducibly follows a plausible cytoplasmic pattern, while treating broad extracellular deposits as suspect (HPA tissue IHC; standard IHC practice).
What should I score when KDELR3 staining varies between specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring compartment before reviewing specimen groups, because HPA describes cytoplasmic, sometimes granular staining (HPA tissue IHC; standard IHC practice). Within a fixed region of interest, report the percentage of positive eligible cells and an intensity-weighted H-score, or measure positive-cell density per mm² when cell abundance is the question (standard IHC practice). Normalise cell-based measures to all eligible cells of the same type, and area-based measures to the analysed viable tissue area (standard IHC practice). Keep retrieval, staining batch, exposure to chromogen, and scoring thresholds consistent, and report HPA's uncertain tissue-IHC reliability alongside any biological comparison (page retrieval specification; standard IHC practice; HPA tissue IHC reliability).
When is KDELR3 staining credible rather than a tissue artefact?
A credible result is reproducible in intact cells, has a cytoplasmic or granular pattern compatible with KDELR3's ER–Golgi trafficking, and exceeds the no-primary control (HPA tissue IHC; UniProt O43731 localisation; standard IHC practice). HPA reports high staining in appendix and small-intestine glandular cells but rates its tissue-IHC reliability uncertain because antibody staining and RNA expression show very low consistency (HPA tissue IHC). Examine unexpected nuclear-only signal, staining confined to section edges or necrotic areas, and signal attributable to endogenous peroxidase before scoring a positive cell (UniProt O43731 localisation; standard IHC practice). Confirm disputed patterns with an independent antibody or orthogonal expression evidence, and describe discordant specimens without assuming that either assay alone establishes specificity (standard IHC practice; HPA tissue IHC reliability).
Boster reagents
Best KDELR3 / ER lumen protein-retaining receptor 3 IHC Antibodies
Anti-KDELR3 catalog entries list IHC for human, mouse, and rat and IF/ICC for human and mouse (catalog applications/reactivity). The supplied image documents paraffin-embedded human breast carcinoma IHC (A14682-1 image caption).
Real IHC data
Anti-KDEL Receptor 3 (L95) KDELR3 Antibody
Cat # A14682-1
A14682-1 is the only card that renders; its IHC image shows paraffin-embedded human breast carcinoma at 1:50 (A14682-1 image caption). Its listed IHC reactivity includes human, mouse, and rat, while the supplied image documents human tissue only (A14682-1 applications/reactivity; A14682-1 image caption).
Which to pick: For paraffin-section IHC, choose A14682-1: its own image documents human breast carcinoma at 1:50; the fixative is unreported (A14682-1 image caption). For IF/ICC, A30677 lists both applications for human and mouse, with no IF image supplied (A30677 applications/reactivity/image alts). For cross-species IHC, A14682-1 has the broadest listed reactivity—human, mouse, and rat—although its image documents human tissue only (A14682-1 reactivity/image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43731 (ERD23_HUMAN, ER lumen protein-retaining receptor 3).
- Human Protein Atlas. KDELR3 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. KDELR3 subcellular location (ICC-IF): Localized to the endoplasmic reticulum..
- Human Protein Atlas. KDELR3 antibody validation summary (2 antibodies).
- KDELR3 overexpression as a novel prognostic and diagnostic biomarker in glioma: comprehensive bioinformatic analysis insights. Scientific reports 2024 — PMC11681132.
- Exploration of the Key Pathways and Genes Involved in Osteoarthritis Genesis: Evidence from Multiple Platforms and Real-World Validation. Journal of inflammation research 2024 — PMC11625429.
- KDELR2 is necessary for chronic obstructive pulmonary disease airway Mucin5AC hypersecretion via an IRE1α/XBP-1s-dependent mechanism. Journal of cellular and molecular medicine 2024 — PMC11451269.
- Melanoblast transcriptome analysis reveals pathways promoting melanoma metastasis. Nature communications 2020 — PMC6965108.
- PubMed PMID:12529303 — UniProt-cited evidence.
- PubMed PMID:15461802 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KDELR3 (ER lumen protein-retaining receptor 3). Expected localisation Cytoplasmic staining, sometimes granular (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A14682-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A14682-1); 2. Fixation: Image fixative and duration unreported (datasheet A14682-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-KDELR3, 1:50-1:200 (datasheet A14682-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A14682-1); Caveat: Very low staining–RNA concordance (HPA tissue IHC); Isoform / epitope: 2 isoforms; map epitopes to lumenal or cytoplasmic loops (UniProt); Regulation: No regulation annotated (UniProt). Evidence: UniProt O43731, HPA tissue IHC · Uncertain, HPA ICC-IF · endoplasmic reticulum., Datasheet A14682-1, PMC11681132, PMC11625429. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KDELR3/immunohistochemistry-ihc-guide.