KRT7 / Keratin, type II cytoskeletal 7 · IHC design guide

Design Immunohistochemistry for KRT7

Plan KRT7 staining in paraffin sections using the catalog antibody's IHC range of 2–5 μg/ml (datasheet A02416-2). Assess cytoplasmic and membranous signal in glandular epithelium against the reported normal tissue pattern (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for KRT7 (IHC for KRT7): expected localisation Cytoplasmic and membranous in glandular epithelium (HPA tissue IHC), antibody A02416-2, validated IHC image, and IHC protocol steps
Printable KRT7 IHC protocol sheet — expected localisation Cytoplasmic and membranous in glandular epithelium (HPA tissue IHC), antibody A02416-2, controls and protocol steps. Open the full KRT7 IHC guide →

KRT7 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 and membranous in glandular epithelium (HPA tissue IHC)
Staining pattern Glandular epithelial cytoplasmic and membranous staining (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet A02416-2)
Positive control ⓘ Breast+4 more · see all
Negative control ⓘ Adipose tissue+4 more · see all
Important caveats
Reasons your staining may differ from the expected pattern.
Fixation Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02416-2)
Caveat Normal glandular epithelium can stain strongly (HPA tissue IHC)
Regulation Tissue-dependent epithelial expression (HPA tissue IHC)
Isoform / epitope No listed isoforms; intracellular epitope access matters (UniProt)
Section 1

Recommended KRT7 IHC & IF Protocols

The catalog antibody protocol uses EDTA retrieval (datasheet A02416-2). The published IHC protocols below provide conditions for urothelial tumours, sarcoma, and cervical carcinoma (PMC2361501; PMC6751227; PMC9131751).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human breast cancer tissue; fixative not specified (datasheet A02416-2)
FixationImage fixative and duration unreported (datasheet A02416-2); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A02416-2); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A02416-2)
Primary antibodyRabbit anti-KRT7, 2-5 μg/ml (datasheet A02416-2)
Primary incubationOvernight at 4 °C (datasheet A02416-2)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet A02416-2)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultKRT7-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression mainly in glandular epithelium. No signal in the no-primary control.
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A02416-2). Evaluate the published retrieval conditions with their respective antibodies (PMC2361501; PMC6751227).
Section 2

What Is the Expected KRT7 Staining Pattern?

KRT7 is a cytoplasmic intermediate filament protein with no transmembrane segment (UniProt P08729: localisation, topology). In paraffin IHC, expect cytoplasmic and sometimes membranous staining, mainly in glandular epithelium (HPA: tissue IHC profile). HPA rates the tissue pattern Enhanced while reporting medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability); interpret signal by cell type and compartment.

What am I looking at on my slide?
Strong cytoplasmic staining in breast glandular cells or bronchial respiratory epithelium.This fits two observed high-expression patterns (HPA: breast glandular cells, High; bronchus respiratory epithelial cells, High). Assess the stained cells within their tissue context, since the HPA observations identify specific cell populations.
Cytoplasmic staining with an apparent membranous component in glandular epithelium.This can fit the reported paraffin IHC profile (HPA: cytoplasmic and membranous expression mainly in glandular epithelium). KRT7 has no transmembrane segment, so a membrane-like edge does not establish membrane insertion (UniProt P08729: topology).
Nuclear-only staining, with little or no cytoplasmic signal in expected positive cells.Treat this as a compartment mismatch requiring investigation: UniProt places KRT7 in the cytoplasm, and HPA supports cytosol and intermediate filaments by ICC-IF (UniProt P08729: localisation; HPA: subcellular).
Signal concentrated in adipocytes or bone-marrow hematopoietic cells.Those cell populations are reported as not detected (HPA: adipocytes, Not detected; bone marrow hematopoietic cells, Not detected). Check for cross-reactivity or endogenous detection activity before scoring such signal as KRT7 (standard IHC practice).
Widespread haze across cells and surrounding tissue, without a discernible cell pattern.Diffuse background cannot establish the reported cell-specific pattern (HPA: tissue IHC profile). Inspect the no-primary control and detection steps for nonspecific signal, then reassess the positive cells (standard IHC practice).
💡Expected KRT7 appearanceA convincing positive shows strong cytoplasmic, sometimes membranous-appearing signal in an HPA high-expression epithelial population, such as breast glandular cells (HPA: tissue IHC profile; breast glandular cells, High); nuclear-only or cell-independent haze is suspect (UniProt P08729: localisation; standard IHC practice).
How each factor affects the staining
Cell compartment and topology (UniProt P08729: localisation, topology).KRT7 is cytoplasmic and lacks a transmembrane segment (UniProt P08729: localisation, topology). Interpret a membrane-like outline alongside cytoplasmic staining; do not infer a membrane-spanning protein from the IHC appearance alone.
Positive tissue and cell choice (HPA: tissue IHC).HPA reports High staining in breast glandular cells, bronchial respiratory epithelial cells, kidney collecting ducts and lung alveolar type I cells (HPA: listed cell-specific levels). Select a documented cell population when judging a positive control.
Negative and low-level comparators (HPA: tissue IHC).HPA lists adipocytes and bone-marrow hematopoietic cells as Not detected, while stomach glandular cells are Low (HPA: listed cell-specific levels). A weak stomach result therefore has a different interpretation from failure in a documented High population.
Antibody evidence (HPA: antibody validation; tissue IHC reliability).HPA007272 and CAB000028 have Enhanced IHC validation; CAB079970 and CAB080115 have Supported IHC validation (HPA: antibody list). HPA also reports medium antibody–RNA consistency for the tissue profile (HPA: reliability description).
IF/ICC: where should signal appear? (HPA: subcellular).Expect cytosol and intermediate-filament localisation in ICC-IF; HPA marks filaments Approved and cytosol Supported (HPA: subcellular). This is an interpretation cue for IF/ICC, whose preparation needs its separate guide.
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
No signal in a documented High population, such as breast glandular cells (HPA: breast, High).The staining run or selected cell population may be unsuitable; HPA's level is cell-specific (HPA: tissue IHC).Confirm that the expected cells are present, then review the positive control, antibody dilution, retrieval and detection run records (standard IHC practice). No KRT7-specific retrieval condition is established by these sources.
Weak staining in stomach glandular cells (HPA: stomach glandular cells, Low).Weak signal may reflect the reported low expression, rather than a failed run (HPA: stomach, Low).Compare with a documented High population on a concurrently processed control, and score the stomach cells in their own context (HPA: tissue IHC; standard IHC practice).
Nuclear-only signal in otherwise expected epithelial cells.The compartment conflicts with cytoplasmic KRT7 localisation (UniProt P08729: localisation; HPA: subcellular).Check the no-primary control, counterstain and detection specificity; require a reproducible cytoplasmic component before accepting the result (standard IHC practice).
Apparent positivity in adipocytes or bone-marrow hematopoietic cells.HPA reports these cells as Not detected; unexpected signal could reflect cross-reactivity or endogenous detection activity (HPA: tissue IHC; standard IHC practice).Compare no-primary and appropriate detection controls, inspect cell identity, and repeat with an independently validated IHC antibody if needed (standard IHC practice; HPA: antibody validation).
Diffuse staining obscures epithelial borders and cell identity.Background from blocking, antibody concentration or detection steps can obscure a cell-specific pattern (standard IHC practice; HPA: tissue IHC profile).Review blocking and washes, compare the no-primary control, and adjust the staining workflow only after locating the background source (standard IHC practice).
Two antibodies give different tissue patterns.HPA assigns different validation levels to listed IHC antibodies, while its tissue profile has medium antibody–RNA consistency (HPA: antibody list; reliability description).Compare both stains in the same documented High and Not detected cell populations, then check each antibody's IHC validation and controls before resolving the discrepancy (HPA: tissue IHC; standard IHC practice).

Sample controls for KRT7 IHC & IF

🧪Run breast first: its glandular cells should show strong KRT7 staining (HPA: Breast glandular cells High). Use adipose tissue as the negative; adipocytes within the breast section, if present, should remain unstained and provide an internal negative comparison (HPA: Adipose tissue adipocytes Not detected).
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KRT7 in HaCaT, SiHa, U2OS, with annotated localisation: Intermediate filaments (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (caption: rabbit anti-KRT7 primary). A KRT7 knockout sample can serve as a biological negative control. For the HRP/DAB method, block endogenous peroxidase and check the breast section for background staining (caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02416-2 tissue-IHC caption does not state the fixative (caption: fixative not stated). The paraffin-section example used 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 are easier (caption: EDTA heat retrieval; HPA: ICC-IF images in HaCaT, SiHa and U2OS). In breast sections, distinguish glandular staining from background in adjacent adipocytes, which are expected to be negative (HPA: Breast glandular cells High; HPA: Adipose tissue adipocytes Not detected).

HPA tissue IHC evidence for KRT7

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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

TissueCell typeLevelEvidenceSource
Breast Glandular cells High Protein (IHC) HPA →
Bronchus Respiratory epithelial cells High Protein (IHC) HPA →
Cervix Glandular cells High Protein (IHC) HPA →
Endometrium Ciliated epithelial cells High Protein (IHC) HPA →
Fallopian tube Glandular cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Caudate Glial cells Not detected Protein (IHC) HPA →
Section 3

Advanced KRT7 IHC Tips

Troubleshoot KRT7 staining in paraffin sections by checking retrieval, cell identity, filament localisation and the controls used for chromogenic detection.

What retrieval should I try first when KRT7 staining is weak in paraffin sections?
Start with heat-mediated EDTA at pH 8.0 for KRT7 IHC in paraffin sections (datasheet A02416-2). The selected tissue example paired that retrieval with 2 μg/ml primary antibody overnight at 4°C, providing a documented starting condition (datasheet A02416-2). If staining remains weak, compare retrieval durations on adjacent sections while holding antibody concentration, detection and development constant (standard IHC practice). Include a known positive epithelial control to distinguish inadequate retrieval from a sample with little detectable KRT7 (HPA: high in breast glandular cells; standard IHC practice). If an alternative buffer is tested, treat it as a fallback and record the change before comparing staining intensity (standard IHC practice).
How should I troubleshoot weak KRT7 staining when tissue fixation varies?
The selected KRT7 caption identifies a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A02416-2). Record the fixative and fixation duration for each specimen, then compare sections processed with the same retrieval and chromogenic detection settings (standard IHC practice). Use heat-mediated EDTA at pH 8.0 and the documented 2 μg/ml primary concentration as starting conditions for that comparison (datasheet A02416-2). If a well-preserved positive control stains and a test specimen does not, assess tissue preservation and processing before changing the KRT7 score (standard IHC practice). Do not attribute the difference to a particular fixation effect without a controlled comparison (standard IHC practice).
Should KRT7 appear on the membrane or in the cytoplasm?
Expect predominantly cytoplasmic, filament-associated KRT7 signal because the protein is an intermediate-filament keratin localised to the cytoplasm (UniProt P08729; HPA subcellular: intermediate filaments). HPA also describes cytoplasmic and membranous staining in glandular epithelium, so assess whether apparent membrane accentuation accompanies convincing cytoplasmic staining (HPA tissue IHC). KRT7 has no transmembrane segment; a crisp membrane-only DAB outline therefore needs comparison with cellular morphology and controls before interpretation (UniProt P08729 topology; standard IHC practice). Examine matched sections at the same DAB development setting, since heavy chromogen deposition can obscure the filament pattern (standard IHC practice). Score staining in the expected epithelial cells separately from surrounding tissue (HPA tissue IHC; standard IHC practice).
Could epitope masking or protein modifications explain uneven KRT7 staining?
The supplied record lists 0 isoforms, so an isoform-specific explanation for uneven staining is unsupported here (UniProt P08729). KRT7 contains an intermediate-filament rod spanning residues 91–403 and reported modified residues, but the antibody epitope is not supplied (UniProt P08729; datasheet A02416-2). Do not infer that a particular modification blocks antibody binding without epitope mapping and a direct comparison (standard IHC practice). Compare adjacent sections using the documented EDTA pH 8.0 retrieval and a consistent antibody concentration before changing conditions (datasheet A02416-2; standard IHC practice). If staining varies within a section, assess tissue preservation and retrieval uniformity alongside cell identity (standard IHC practice).
How can I check a KRT7 IHC pattern with multiplex immunofluorescence?
Use an epithelial marker appropriate to the expected positive cell type when comparing KRT7 with cell identity in multiplex IF (HPA tissue IHC: glandular and respiratory epithelial staining; standard IF practice). Choose a fluorophore in a channel with low measured tissue autofluorescence, and include single-stain and unstained controls for channel assessment (standard IF practice). KRT7 is cytoplasmic and has no transmembrane segment, so permeabilise cells sufficiently for intracellular antibody access while preserving filament morphology (UniProt P08729 topology; HPA subcellular; standard IF practice). The antibody epitope and an IF protocol for this catalog antibody are not provided, so optimise permeabilisation empirically (datasheet A02416-2). Keep IF observations distinct from the paraffin-section chromogenic IHC result when comparing patterns (datasheet A02416-2; standard IF practice).
How do I reduce diffuse DAB background without losing KRT7 signal?
First compare the stained section with a no-primary control to locate signal arising from the detection system (standard IHC practice). Apply a peroxidase block before HRP detection, then check washing, blocking and DAB development when diffuse colour persists (standard IHC practice). The selected KRT7 example used 10% goat serum, 2 μg/ml primary antibody and an HRP/DAB workflow; these are documented starting conditions (datasheet A02416-2). Titrate primary antibody or shorten DAB development one variable at a time while retaining a known positive epithelial control (HPA: high in breast glandular cells; standard IHC practice). Judge improvement by the separation between epithelial cytoplasmic signal and background (UniProt P08729 localisation; standard IHC practice).
How should I quantify heterogeneous KRT7 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the epithelial compartment before scoring, because HPA reports high KRT7 staining in selected epithelial cell populations (HPA tissue IHC). For each section, report the percentage of positive target cells and intensity categories, or calculate an H-score from intensity-weighted percentages on a 0–300 scale (standard IHC practice). If counting discrete positive cells, express density per mm² of evaluable target tissue rather than total section area (standard IHC practice). Keep retrieval, antibody concentration, DAB development and image thresholds consistent across compared sections (standard IHC practice). Record excluded damaged or necrotic regions and normalise counts to the assessed epithelial area or cell number (standard IHC practice).
How can I distinguish genuine KRT7 positivity from staining artefact?
Prioritise cytoplasmic, filament-compatible staining in the expected epithelial cells when calling KRT7 positive (UniProt P08729; HPA subcellular; HPA tissue IHC). Compare suspect cells with a positive epithelial control and a no-primary control, especially when staining appears outside the expected compartment (HPA tissue IHC; standard IHC practice). Strong signal confined to tissue edges or necrotic regions needs review against morphology and adjacent intact tissue (standard IHC practice). Check a no-primary section for residual endogenous enzyme signal before attributing brown DAB deposits to KRT7 (standard IHC practice). HPA assigns the tissue staining profile Enhanced reliability with medium RNA–staining consistency, so interpret discordant cases with that limitation in mind (HPA tissue IHC).
Boster reagents

Best KRT7 / Keratin, type II cytoskeletal 7 IHC Antibodies

Anti-KRT7 antibodies have IHC images from human breast cancer, prostate and uterus, and mouse and rat liver (catalog IHC captions); IF images show human cancer sections and HeLa cells (catalog IF captions).

Real IHC data IHC analysis of KRT7 using anti-KRT7 antibody (A02416-2). KRT7 was detected in a paraffin-embedded section of human breast 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-KRT7 Antibody (A02416-2) 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.
Anti-KRT7 Antibody ®
Cat # A02416-2
Real IHC data Human prostate was stained with anti-Cytokeratin 7 rabbit antibody
Anti-Cytokeratin 7 Rabbit Monoclonal Antibody
Cat # M02416-4
Real IHC data Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Cytokeratin 7 Antibody .
Anti-Cytokeratin 7 KRT7 Rabbit Monoclonal Antibody
Cat # M02416-1
Real IHC data Immunohistochemical analysis of paraffin-embedded human uterus, using Cytokeratin 7 Antibody.
Anti-Cytokeratin 7 KRT7 Rabbit Monoclonal Antibody
Cat # M02416-2

A02416-2 shows IHC in human breast cancer and IF in human cancer sections (A02416-2 image captions); M02416-4 shows IHC in human prostate and mouse and rat liver (M02416-4 IHC captions). M02416-1 shows IHC in human breast carcinoma and IF in HeLa cells (M02416-1 image captions); M02416-2 shows IHC in human uterus and IF in HeLa cells (M02416-2 image captions).

Which to pick: For tissue IHC, A02416-2 provides a documented starting point in paraffin human breast cancer sections: EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody overnight at 4°C; the fixative is unreported (A02416-2 IHC caption). For human IF/ICC, choose rabbit monoclonal M02416-1: IF is pictured in HeLa cells, and IF and ICC are listed applications (M02416-1 IF caption; catalog applications and clonality). For IHC across species, choose rabbit monoclonal M02416-4, with images from human prostate and mouse and rat liver (M02416-4 IHC captions; catalog clonality); its tissue processing and fixatives are unreported (M02416-4 IHC captions).

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

References

  1. UniProt Consortium. UniProt entry P08729 (K2C7_HUMAN, Keratin, type II cytoskeletal 7).
  2. Human Protein Atlas. KRT7 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. KRT7 subcellular location (ICC-IF): Localized to the cytosol and intermediate filaments..
  4. Human Protein Atlas. KRT7 antibody validation summary (5 antibodies).
  5. Gene expression profiling of noninvasive primary urothelial tumours using microarrays. British journal of cancer 2005 — PMC2361501.
  6. Differential gene expression identifies KRT7 and MUC1 as potential metastasis-specific targets in sarcoma. Cancer management and research 2019 — PMC6751227.
  7. KRT7 Overexpression is Associated with Poor Prognosis and Immune Cell Infiltration in Patients with Pancreatic Adenocarcinoma. International journal of general medicine 2021 — PMC8233003.
  8. ALDH1 & CD133 in invasive cervical carcinoma & their association with the outcome of chemoradiation therapy. The Indian journal of medical research 2021 — PMC9131751.
  9. PubMed PMID:2415537 — UniProt-cited evidence.
  10. PubMed PMID:2459129 — UniProt-cited evidence.
  11. PubMed PMID:10492017 — UniProt-cited evidence.