This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
CRK / Adapter molecule crk · IHC design guide
Design Immunohistochemistry for CRK
Plan paraffin-section CRK staining around its membranous and cytoplasmic tissue pattern (HPA tissue IHC). Use high-staining glandular cells as a reference and assess possible off-target staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CRK 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 | Membranous and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular cells: membranous and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9101) | |
| Positive control | Duodenum+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) | |
| Caveat | Presumed off-target binding can mimic CRK staining (HPA tissue IHC) | |
| Regulation | Cell adhesion shifts CRK to the membrane (UniProt) | |
| Isoform / epitope | 2 isoforms (Crk-I/II); check antibody epitope coverage (UniProt) |
Section 1
Recommended CRK IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet PB9101). The published protocol below describes CRK IHC in human nephroblastoma and adjacent tissue (PMC11896132).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PB9101) |
| Fixation | Image fixative and duration unreported (datasheet PB9101); 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 PB9101); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9101) |
| Primary antibody | Rabbit anti-CRK, 2-5μg/ml (datasheet PB9101) |
| Primary incubation | Overnight at 4 °C (datasheet PB9101) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9101) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CRK-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Membranous and cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet PB9101). Citrate buffer was used with a different CRK antibody (PMC11896132).
Section 2
What Is the Expected CRK Staining Pattern?
CRK is a cytoplasmic protein that can move to the plasma membrane during cell adhesion; it has no transmembrane segment (UniProt P46108). In paraffin sections, expect membranous and cytoplasmic staining, especially in duodenal glandular cells, kidney tubular cells, and hepatocytes (HPA tissue IHC). HPA rates its tissue staining Approved, with medium agreement between staining and RNA data and presumed off-target binding disregarded (HPA tissue IHC).
| Clear cytoplasmic staining, with some membrane accentuation, in kidney tubular cells or hepatocytes. | This fits the reported tissue pattern: both cell populations stain at a High level, and HPA describes membranous and cytoplasmic expression across several tissues (HPA tissue IHC). Score the named cells and compartments separately from nearby structures. |
| Strong staining confined to nuclei, with little cytoplasmic or membrane signal. | Treat a nuclear-only IHC pattern as questionable: UniProt places CRK in cytoplasm and at cell membrane (UniProt P46108). HPA reports nucleoplasm as an uncertain additional location in ICC-IF, so nuclear signal alone cannot confirm specific IHC staining (HPA subcellular). |
| Apparent positive staining in adipocytes while expected positive cells remain weak. | Adipocytes in adipose tissue and breast were Not detected in HPA's listed observations (HPA tissue IHC). Check antibody specificity and endogenous detection activity before assigning this unexpected cell staining to CRK (standard IHC practice). |
| Broad, even color covers cells and surrounding section with little cell-specific contrast. | A diffuse field does not reproduce HPA's cell-resolved membranous and cytoplasmic pattern (HPA tissue IHC). Compare with a no-primary control and inspect blocking, washes, and chromogen development for general background (standard IHC practice). |
| No staining in duodenal glandular cells, kidney tubular cells, or hepatocytes. | These are High-staining reference populations in the supplied tissue observations (HPA tissue IHC). Verify tissue preservation, retrieval and detection with controls before interpreting an absent signal as low CRK expression (standard IHC practice). |
💡Expected CRK appearanceCall a result consistent with CRK when High-level cytoplasmic staining, sometimes with membrane accentuation, appears in the specified duodenal glandular cells, kidney tubular cells, or hepatocytes; diffuse color or staining restricted to HPA-listed Not detected cell populations warrants review (HPA tissue IHC; UniProt P46108).
| Compartment and topology | CRK is annotated in cytoplasm and at cell membrane, with membrane translocation upon cell adhesion and no transmembrane segment (UniProt P46108). Membrane accentuation can therefore vary; assess it alongside cytoplasmic staining. |
| Reference cell population | HPA lists High staining in duodenal and small-intestinal glandular cells, kidney tubular cells, hepatocytes, and salivary glandular cells (HPA tissue IHC). Its Not detected entries name particular cell populations, not entire tissues. |
| Antibody evidence | The supplied IHC antibody CAB010485 is Approved, while HPA describes overall tissue staining as medium in consistency with RNA and notes presumed off-target binding (HPA antibodies; HPA tissue IHC). Use morphology and controls when interpreting unexpected staining. |
| Isoforms and epitope | UniProt lists Crk-I and Crk-II isoforms and SH2 and SH3 domains (UniProt P46108). The payload gives no IHC antibody epitope, so it cannot establish which isoform the IHC signal represents. |
| IF/ICC Q: Does its pattern match IHC? | HPA places CRK mainly at the plasma membrane in ICC-IF, with approved additional actin-filament and basal-body locations; nucleoplasm is uncertain (HPA subcellular). Interpret those observations in the separate IF/ICC guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| A listed High-staining cell population is blank. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Check a known-positive section alongside the sample, then verify the validated IHC workflow, retrieval, primary-antibody step, and detection controls (standard IHC practice). |
| The whole section shows diffuse chromogen. | Background from endogenous enzyme activity, insufficient blocking or washing, or prolonged development can obscure cell-specific signal (standard IHC practice). | Compare a no-primary control; address endogenous activity and blocking, improve washes, and review development time (standard IHC practice). |
| Nuclear staining dominates with no convincing cytoplasm. | This differs from UniProt's cytoplasmic and membrane annotation; the additional nucleoplasmic ICC-IF location is uncertain (UniProt P46108; HPA subcellular). | Recheck morphology and the no-primary control, then assess staining in an HPA High-staining cell population before calling the nuclear pattern specific (HPA tissue IHC; standard IHC practice). |
| An HPA-listed Not detected population appears strongly positive. | Unexpected staining could reflect cross-reactivity or endogenous detection activity; HPA reports presumed off-target binding in its tissue assessment (HPA tissue IHC; standard IHC practice). | Confirm the exact cell type, compare an HPA High-staining population and a no-primary control, and interpret the unexpected signal cautiously (HPA tissue IHC; standard IHC practice). |
| Membrane staining is absent but cytoplasmic staining is clear. | CRK is also cytoplasmic, and UniProt notes membrane translocation upon cell adhesion (UniProt P46108). | Assess the complete cell pattern and positive-reference population; do not require a continuous membrane outline to recognize a cytoplasmic result consistent with HPA tissue IHC (HPA tissue IHC; UniProt P46108). |
| The result is reported as specific to Crk-I or Crk-II. | Both isoforms are listed, but the supplied antibody record gives no epitope or isoform-selectivity evidence (UniProt P46108; HPA antibodies). | Report staining as CRK unless independent isoform-specific validation is available; do not infer isoform identity from compartment or intensity alone (UniProt P46108; standard IHC practice). |
Experiment evidence
IHC & IF images showing CRK staining
PB9101 · IHCIHC analysis of Crk p38 using anti-Crk p38 antibody (PB9101). Crk p38 was detected in a paraffin-embedded section of human lung cancer tissu
PB9101 · IHCIHC analysis of Crk p38 using anti-Crk p38 antibody (PB9101). Crk p38 was detected in a paraffin-embedded section of mouse brain tissue. Hea
M02533-3 · IFIF analysis of CRKII using anti-CRKII antibody (M02533-3) and anti-Beta Tubulin antibody (M01857-3). CRKII was detected in immunocytochemica
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000167193-CRK/tissue/cerebral+cortex#imid_3638020
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000167193-CRK/tissue/colon#imid_3637930
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000167193-CRK/tissue/liver#imid_3637939
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000167193-CRK/tissue/kidney#imid_3637948
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA068087; green = CRK, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA068087; green = CRK, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CRK IHC & IF
🧪Run duodenum first and expect staining in its glandular cells (HPA: High in duodenal glandular cells). Use adipose tissue as the negative tissue, with adipocytes at background (HPA: Not detected in adipocytes); on the duodenum slide, treat cells without specific staining as internal background references, without assuming a particular non-glandular cell type is CRK-negative.
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CRK in A-549, HaCaT, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a matched rabbit IgG isotype control appropriate to the primary antibody’s clonality; and, if available, a CRK knockout specimen or validated peptide-block control (caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase in duodenal sections before chromogenic detection and assess endogenous biotin background if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9101 paraffin-section caption does not state the fixative (PB9101 caption). The reported IHC procedure uses heat-mediated retrieval in EDTA at pH 8.0; this supports starting with that retrieval condition but does not establish that retrieval is required for every specimen (PB9101 caption). Paraffin-section IHC has the more direct supplied protocol evidence; frozen-section performance is unreported, while HPA ICC-IF images document cellular localization without establishing an easier workflow (PB9101 caption; HPA: ICC-IF images).
HPA tissue IHC evidence for CRK
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Presumed 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CRK IHC Tips
Use the catalog antibody’s tissue IHC conditions as a starting point, then assess CRK staining by compartment, cell type, and matched controls.
What retrieval conditions should I start with for CRK in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet PB9101). The selected paraffin-section example used this retrieval before incubating with 2 μg/ml primary antibody overnight at 4°C, so keep those conditions together for the first comparison (datasheet PB9101). If staining is weak, vary heating duration on matched sections and check tissue integrity and background after each change (standard IHC practice). Interpret improvement against cytoplasmic or membranous staining in expected cells, since CRK is reported in those compartments and retrieval can also increase nonspecific staining (UniProt P46108 localisation; HPA: tissue IHC profile; standard IHC practice).
Could fixation explain weak or uneven CRK staining?
Target-specific fixation sensitivity for CRK is unknown from the supplied evidence; the selected image describes a paraffin-embedded section but does not state its fixative (datasheet PB9101 caption). Record the fixative, fixation duration, tissue thickness, and processing history for each specimen before comparing staining across batches (standard IHC practice). On serial sections, hold retrieval at EDTA pH 8.0 and primary antibody at 2 μg/ml while checking whether weak areas track processing differences (datasheet PB9101; standard IHC practice). Use a consistently processed reference section and inspect morphology alongside staining, because damaged or poorly preserved tissue can mislead chromogenic scoring (standard IHC practice).
Where should CRK staining appear, and how should I assess nuclear signal?
Expect cytoplasmic and membranous tissue staining, with membrane recruitment possible during cell adhesion (HPA: tissue IHC profile; UniProt P46108 localisation). In matched sections, compare membrane and cytoplasmic intensity within the same cell population rather than treating every dark cell border as equivalent (standard IHC practice). HPA reports mainly plasma-membrane localisation by subcellular imaging and an additional nucleoplasmic location marked uncertain, so nuclear DAB alone needs independent support (HPA: subcellular localisation). Check whether nuclear colour survives omission of the primary antibody and whether it follows intact cell boundaries rather than folds, pigment, or damaged regions (standard IHC practice).
Can this antibody distinguish Crk-I from Crk-II in tissue?
CRK has 2 reported isoforms, Crk-I and Crk-II, but the supplied evidence does not map the catalog antibody’s epitope to either isoform (UniProt P46108 isoforms; datasheet PB9101 caption). Do not assign a stained compartment or cell population to one isoform from this chromogenic result alone (standard IHC interpretation). UniProt places the SH2 domain at residues 13–118 and SH3 domains at 132–192 and 235–296; these coordinates do not establish which sequence the antibody recognises (UniProt P46108 domains). For an isoform-specific conclusion, first obtain epitope documentation or independent isoform-specific validation, then compare matched sections using identical processing and scoring (standard IHC practice).
How can I use IF to investigate an ambiguous CRK IHC pattern?
Use IF as a separate follow-up to the paraffin-section DAB result, whose documented conditions are EDTA pH 8.0 retrieval and 2 μg/ml catalog antibody (datasheet PB9101). Multiplex CRK with a validated marker for the cell population under study; hepatocytes offer a tissue IHC comparison because HPA reports high CRK staining there (HPA: high in hepatocytes; standard IF practice). Choose fluorophores after checking the specimen’s autofluorescence and include single-colour controls to distinguish overlap from bleed-through (standard IF practice). CRK lacks a transmembrane segment and is reported in the cytoplasm and at the membrane, so use appropriate permeabilisation for intracellular epitopes while recognising that this antibody’s epitope side is unspecified (UniProt P46108 topology and localisation; standard IF practice).
How do I reduce diffuse DAB background without losing CRK signal?
Begin with the documented 10% goat-serum block and 2 μg/ml primary antibody, then compare serial sections while changing one variable at a time (datasheet PB9101; standard IHC practice). Include a primary-omission section to reveal secondary-reagent or chromogen background, and apply a peroxidase block before HRP and DAB detection (standard IHC practice). The selected example used a peroxidase-conjugated secondary for 30 minutes at 37°C and DAB development, so monitor development time and washing when background rises (datasheet PB9101; standard IHC practice). Score retained staining only when it follows plausible cytoplasmic or membranous cell patterns rather than diffuse colour across stroma or section edges (HPA: tissue IHC profile; standard IHC practice).
How should I quantify heterogeneous CRK staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because CRK tissue staining is reported as membranous and cytoplasmic across several tissues (HPA: tissue IHC profile). For an intensity-based readout, report an H-score from 0–300 using the percentage of cells at each intensity, and keep membrane and cytoplasm scores separate (standard IHC practice). Also report the percentage of positive cells, or positive-cell density per mm² when tissue area varies; normalise to eligible, intact cells or analysed viable tissue area, respectively (standard IHC practice). Apply one threshold and imaging setup across batches, with a consistently processed reference section to detect drift (standard IHC practice).
What distinguishes a credible CRK-positive cell from a staining artefact?
A credible result places DAB in intact cells with cytoplasmic or membranous staining and agrees with an appropriate tissue reference (HPA: tissue IHC profile; standard IHC practice). HPA reports high staining in kidney tubule cells and hepatocytes, but also notes medium staining–RNA consistency and presumed off-target binding, so these references require cautious interpretation (HPA: tissue IHC). Treat isolated nuclear-only staining as uncertain because the HPA nucleoplasmic location is marked uncertain, and inspect unexpected cell types before assigning biology (HPA: subcellular localisation; standard IHC practice). Exclude section-edge concentration, necrotic areas, pigment, and residual endogenous peroxidase using morphology and primary-omission controls before scoring positives (standard IHC practice).
Boster reagents
Best CRK / Adapter molecule crk IHC Antibodies
The catalog includes tissue IHC images from human lung cancer and mouse and rat brain (PB9101 IHC captions), plus an IF image from HeLa cells (M02533-3 IF caption).
Real IHC data
Anti-Crk p38 Antibody ®
Cat # PB9101
Real IF data
Anti-CRKII Rabbit Monoclonal Antibody
Cat # M02533-3
PB9101 will render with an IHC image from a paraffin-embedded human lung cancer section; its other IHC captions document mouse and rat brain sections (PB9101 IHC captions). M02533-3 will render with an IF image from HeLa cells (M02533-3 IF caption).
Which to pick: Choose PB9101 for chromogenic IHC on paraffin sections: its IHC caption documents that preparation, while the fixative is unreported (PB9101 IHC caption). Choose M02533-3 for IF/ICC: it is a rabbit monoclonal, lists IF and ICC applications, and has a HeLa IF image (M02533-3 catalog and IF caption). For work across human, mouse, and rat samples, both list reactivity with all three species; PB9101 additionally has IHC images from each, while M02533-3 has an IF image from human cells (catalog reactivity; PB9101 IHC captions; M02533-3 IF caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P46108 (CRK_HUMAN, Adapter molecule crk).
- Human Protein Atlas. CRK tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CRK subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the nucleoplasm, actin filaments and basal body..
- Human Protein Atlas. CRK antibody validation summary (2 antibodies).
- MiR-194-5p inhibited metastasis and EMT of nephroblastoma cells through targeting Crk. The Kaohsiung journal of medical sciences 2020 — PMC11896132.
- Significance of KRAS/PAK1/Crk pathway in non-small cell lung cancer oncogenesis. BMC cancer 2015 — PMC4477307.
- Adaptor protein CRK induces epithelial-mesenchymal transition and metastasis of bladder cancer cells through HGF/c-Met feedback loop. Cancer science 2015 — PMC4471787.
- Increased cellular distribution of vimentin and ret in the cingulum of rat offspring after developmental exposure to decabromodiphenyl ether or 1,2,5,6,9,10-hexabromocyclododecane. Journal of toxicologic pathology 2013 — PMC3695334.
- PubMed PMID:1630456 — UniProt-cited evidence.
- PubMed PMID:8378094 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CRK (Adapter molecule crk). Expected localisation Membranous and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9101). Validated IHC antibody: PB9101 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PB9101); 2. Fixation: Image fixative and duration unreported (datasheet PB9101); 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 retrieval: EDTA pH 8.0 (datasheet PB9101); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9101); 8. Primary antibody: Rabbit anti-CRK, 2-5μg/ml (datasheet PB9101); 9. Primary incubation: Overnight at 4 °C (datasheet PB9101). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Presumed off-target binding can mimic CRK staining (HPA tissue IHC); Isoform / epitope: 2 isoforms (Crk-I/II); check antibody epitope coverage (UniProt); Regulation: Cell adhesion shifts CRK to the membrane (UniProt). Evidence: UniProt P46108, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet PB9101, PMC11896132. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CRK/immunohistochemistry-ihc-guide.