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SGK1 / Serine/threonine-protein kinase Sgk1 · IHC design guide
Design Immunohistochemistry for SGK1
Plan SGK1 paraffin-section IHC around the general cytoplasmic tissue pattern (HPA tissue IHC). This guide covers fixation consistency, antibody selection and interpretation of localisation that can change with cell cycle or stimuli (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SGK1 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 | Predominantly cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00673-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Soft tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Cell cycle and stimuli can shift SGK1 localisation (UniProt) | |
| Regulation | Staining intensity varies by tissue (HPA tissue IHC) | |
| Isoform / epitope | 5 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended SGK1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet A00673-2). The published protocols below cover human paraffin sections and mouse paraffin or frozen sections (PMC6545623; PMC13607775; PMC6877960; PMC10671719).
| Sample | Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A00673-2) |
| Fixation | Image fixative and duration unreported (datasheet A00673-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A00673-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00673-2) |
| Primary antibody | Rabbit anti-SGK1, 0.5-1μg/ml (datasheet A00673-2) |
| Primary incubation | Overnight at 4 °C (datasheet A00673-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A00673-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SGK1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A00673-2). Adjust other steps to the specimen and antibody used in each published protocol.
Section 2
What Is the Expected SGK1 Staining Pattern?
SGK1 is mainly cytoplasmic in tissue IHC, with High staining reported in glandular, hematopoietic and neuronal cells in selected tissues (HPA tissue IHC). Nuclear or membrane-associated signal can be biologically plausible in context (UniProt O00141 localization). SGK1 has no transmembrane segment (UniProt O00141 topology). HPA rates its tissue IHC profile Approved, while noting low consistency between antibody staining and RNA expression (HPA tissue IHC).
| Granular or diffuse cytoplasmic staining in colon or breast glandular cells. | Consistent with the general cytoplasmic profile and High staining in those cells (HPA tissue IHC). Compare intensity with adjacent tissue and controls (standard IHC practice). |
| Nuclear staining accompanies cytoplasmic staining in otherwise plausible positive cells. | SGK1 can enter the nucleus after phosphorylation or growth factor stimulation (UniProt O00141 localization). Interpret nuclear signal alongside cell context and controls (standard IHC practice). |
| Strong extracellular, luminal or tissue-edge deposit dominates while expected cells remain unstained. | This does not match the intracellular SGK1 locations reported by UniProt (UniProt O00141 localization). Suspect a staining artefact and inspect the negative control (standard IHC practice). |
| Soft-tissue fibroblasts stain strongly while the intended positive cells do not. | HPA reports SGK1 as not detected in soft-tissue fibroblasts (HPA tissue IHC). Check for cross-reactivity or endogenous chromogen activity before calling those cells positive (standard IHC practice). |
| No signal appears in a section expected to contain positive glandular cells. | HPA reports High staining in colon and breast glandular cells (HPA tissue IHC). A negative result there calls for control and detection checks; expression can vary by specimen (standard IHC practice). |
💡Expected SGK1 appearanceCall a result positive when the appropriate cells show predominantly cytoplasmic staining, potentially with contextual nuclear signal (HPA tissue IHC; UniProt O00141 localization); High signal is reported in colon and breast glandular cells, whereas strong extracellular deposit is suspect (HPA tissue IHC; standard IHC practice).
| Tissue and cell type | HPA reports High staining in colon glandular cells and bone-marrow hematopoietic cells, but none in soft-tissue fibroblasts (HPA tissue IHC). Compare like cell types (standard IHC practice). |
| Biological state | SGK1 localization varies with cell cycle, hormones and stress; phosphorylation can favor nuclear localization (UniProt O00141 localization). A nuclear component needs context (standard IHC practice). |
| Antibody evidence | Two listed antibodies have Approved IHC status; HPA also reports low consistency between staining and RNA expression (HPA antibodies; HPA tissue IHC). Treat unexpected patterns cautiously (standard IHC practice). |
| Isoforms and epitope | UniProt lists 5 SGK1 isoforms and a kinase domain at residues 98–355 (UniProt O00141). Without an antibody epitope, isoform coverage cannot be inferred (standard IHC interpretation). |
| Topology and processing | SGK1 has no transmembrane segment, signal peptide, propeptide or reported glycosylation sites (UniProt O00141 topology and processing). Exclusive extracellular or luminal staining needs scrutiny (standard IHC practice). |
| Retrieval and detection | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive glandular cells show no chromogenic signal. | The run may have inadequate retrieval, primary-antibody exposure or detection (standard IHC practice); colon glandular cells have High reported staining (HPA tissue IHC). | Review the positive control, retrieval, antibody dilution and detection reagents together; change one condition at a time (standard IHC practice). |
| The entire section has diffuse brown background. | Nonspecific binding, incomplete blocking or endogenous detection activity can obscure cellular staining (standard IHC practice). | Inspect a no-primary control; adjust blocking, antibody concentration and washes, then address endogenous activity for the detection chemistry used (standard IHC practice). |
| Staining is confined to lumens or section edges. | This distribution conflicts with reported intracellular SGK1 locations and may reflect trapped reagent or edge artefact (UniProt O00141 localization; standard IHC practice). | Compare intact interior regions with the no-primary control and repeat after checking wash and detection steps (standard IHC practice). |
| Strong staining appears in soft-tissue fibroblasts. | HPA reports SGK1 not detected in those cells; cross-reactivity or endogenous chromogen activity is possible (HPA tissue IHC; standard IHC practice). | Check a no-primary control and cell identity before assigning SGK1 positivity; compare an appropriate positive tissue in the same run (standard IHC practice). |
| Nuclear signal seems inconsistent with cytoplasmic tissue staining. | SGK1 can shuttle into nuclei, while HPA summarizes tissue IHC as generally cytoplasmic (UniProt O00141 localization; HPA tissue IHC). | Score nuclear and cytoplasmic compartments separately and assess cellular context and controls before rejecting the signal (standard IHC practice). |
| IF/ICC shows nuclear speckles: should I expect that pattern in paraffin IHC? | HPA reports enhanced nuclear-speckle localization in ICC-IF, while its tissue IHC summary is generally cytoplasmic (HPA subcellular; HPA tissue IHC). | Use the tissue IHC profile to interpret the paraffin section; assess any nuclear component with cell context and controls (HPA tissue IHC; UniProt O00141 localization; standard IHC practice). |
Experiment evidence
IHC & IF images showing SGK1 staining
A00673-2 · IHCIHC analysis of SGK1 using anti-SGK1 antibody (A00673-2). SGK1 was detected in paraffin-embedded section of human mammary cancer tissue. Hea
A00673-2 · IHCIHC analysis of SGK1 using anti-SGK1 antibody (A00673-2). SGK1 was detected in paraffin-embedded section of human mammary cancer tissue. Hea
A00673-2 · IFIF analysis of SGK1 using anti-SGK1 antibody (A00673-2). SGK1 was detected in immunocytochemical section of A549 cells. Enzyme antigen retri
M00673 · IHCIHC analysis of SGK1 using anti-SGK1 antibody (M00673). SGK1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat
M00673 · IHCIHC analysis of SGK1 using anti-SGK1 antibody (M00673). SGK1 was detected in a paraffin-embedded section of mouse ovary tissue. Heat mediate
A00673 · IHCIHC analysis of SGK1 using anti-SGK1 antibody (A00673). SGK1 was detected in paraffin-embedded section of human rectal cancer tissue. Heat m
A00673 · IHCIHC analysis of SGK1 using anti-SGK1 antibody (A00673). SGK1 was detected in paraffin-embedded section of human rectal cancer tissue. Heat m
A00673 · IFIF analysis of SGK1 using anti-SGK1 antibody (A00673). SGK1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retriev
A00673-1 · IHCImmunohistochemistry of SGK1 in human stomach tissue with SGK1 antibody at 5 μg/mL.
A00673-1 · IFImmunofluorescence of SGK1 in human stomach tissue with SGK1 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000118515-SGK1/tissue/cerebral+cortex#imid_5743124
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000118515-SGK1/tissue/colon#imid_5743058
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000118515-SGK1/tissue/liver#imid_5743059
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000118515-SGK1/tissue/kidney#imid_5743060
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB022085; green = SGK1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB022085; green = SGK1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SGK1 IHC & IF
🧪Run adrenal gland first: its glandular cells should stain for SGK1 (HPA: High in adrenal gland glandular cells). Use soft tissue fibroblasts as the negative tissue (HPA: Not detected in soft tissue fibroblasts); on the adrenal slide, treat unstained neighboring cells as a background reference, without assuming that every non-glandular cell is SGK1-negative (HPA: High in glandular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Soft tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SGK1 in A-431, U-251MG, U2OS, HeLa, NIH 3T3, with annotated localisation: Nuclear speckles (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and concentration-matched rabbit IgG isotype control, plus an SGK1 knockout specimen or a validated peptide-block control (caption: rabbit primary; standard IHC practice). Check endogenous peroxidase and biotin background in the adrenal section because the selected detection uses biotin, streptavidin and DAB (caption: biotinylated secondary and SABC–DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A00673-2 caption does not state the fixative (caption: fixative not stated). The demonstrated paraffin-section workflow uses heat retrieval in EDTA at pH 8.0, but the evidence does not establish whether SGK1 staining depends on that retrieval condition (caption: EDTA heat retrieval). Frozen sections and IF cannot be ranked as easier from this IHC evidence; in adrenal tissue, compare DAB staining with the no-primary slide to assess pigment-like background (caption: adrenal tissue choice from HPA; standard IHC practice).
HPA tissue IHC evidence for SGK1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SGK1 IHC Tips
Troubleshoot SGK1 staining in paraffin sections by checking retrieval, cellular compartment, detection background, and matched tissue controls.
How should I retrieve SGK1 antigen when staining paraffin sections?
Use heat-mediated EDTA at pH 8.0 for SGK1 staining in paraffin sections (datasheet A00673-2). The catalog antibody was used at 1 μg/mL overnight at 4°C after this retrieval in human mammary cancer tissue (datasheet A00673-2). If staining is weak, adjust heating duration in small steps while keeping the buffer and pH fixed, and compare sections processed together (standard IHC practice). Check that morphology remains intact and that staining appears in the expected cell population; breast glandular cells show high SGK1 staining in the tissue atlas (HPA: breast glandular cells, High).
Could fixation explain weak or uneven SGK1 staining?
SGK1-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state which fixative was used (datasheet A00673-2). Record each specimen’s fixative and fixation interval, then compare sections with the same processing history before attributing a staining difference to biology (standard IHC practice). Keep section thickness, heat retrieval, and detection conditions consistent across the comparison; use EDTA at pH 8.0 as the documented retrieval starting point (datasheet A00673-2). If signal varies within a section, inspect tissue preservation and include a comparably processed control before changing the antibody concentration (standard IHC practice).
Should SGK1 stain cytoplasm, nuclei, or cell membranes in tissue?
Score cytoplasmic staining first in paraffin sections because the tissue atlas describes general cytoplasmic SGK1 expression (HPA: tissue IHC profile). Nuclear staining can be plausible: SGK1 shuttles between nucleus and cytoplasm with cell cycle and stimulation, and phosphorylation can accompany nuclear localization (UniProt O00141: subcellular location). Membrane-adjacent signal is also plausible in contexts involving epithelial sodium channels, where SGK1 can occupy the cytoplasmic membrane surface (UniProt O00141: subcellular location). Record compartment and cell type separately, and compare adjacent sections rather than calling every nuclear or membranous deposit positive; the atlas also reports nuclear speckles in cellular imaging (HPA: subcellular location).
Can this antibody distinguish SGK1 isoforms or phosphorylation states in IHC?
Do not assign an isoform from this stain without epitope mapping: SGK1 has 5 reported isoforms, but the supplied antibody caption gives no recognized sequence (UniProt O00141: isoforms; datasheet A00673-2). Its kinase domain spans residues 98–355, and its AGC kinase C-terminal domain spans 356–431 (UniProt O00141: domains). Reported phosphorylation sites include Thr256 and Ser397, but the caption does not establish phosphorylation-specific recognition (UniProt O00141: modified residues; datasheet A00673-2). For paraffin-section comparisons, describe the measured signal as catalog-antibody SGK1 immunoreactivity and seek epitope or phospho-specific validation before making narrower claims (standard IHC practice).
How should I assess SGK1 localisation in a multiplex IF experiment?
Plan IF as a separate validation: the catalog evidence here documents chromogenic staining in paraffin sections, whereas nuclear-speckle localisation comes from cellular imaging (datasheet A00673-2; HPA: subcellular location). Pair SGK1 with a marker for the expected cell population, such as a glandular epithelial marker when examining breast glands (HPA: breast glandular cells, High; standard IF practice). Choose a far-red fluorophore if tissue autofluorescence compromises shorter wavelengths, and include single-label controls to check spectral bleed-through (standard IF practice). Permeabilise sufficiently to access cytoplasmic and nuclear epitopes; SGK1 has no transmembrane segment, although it can associate with the cytoplasmic membrane surface (UniProt O00141: topology and subcellular location).
What should I check when SGK1 DAB staining appears diffuse?
First compare the primary-antibody section with a matched no-primary control, then inspect whether deposits track tissue edges, damaged areas, or the expected cells (standard IHC practice). The documented workflow used 10% goat serum, a biotinylated secondary antibody, streptavidin–biotin detection, and DAB (datasheet A00673-2). Check the peroxidase block and a detection-only control if brown signal persists, because endogenous enzyme activity and biotin-dependent detection can contribute background (standard IHC practice). Adjust blocking, washing, and primary concentration systematically while retaining a stained control; general cytoplasmic expression makes diffuse colour alone insufficient evidence of specificity (HPA: tissue IHC profile; standard IHC practice).
How can I quantify SGK1 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; the tissue atlas reports high staining in breast glandular cells and no detected staining in soft-tissue fibroblasts (HPA: tissue IHC). For each annotated region, record percentage of positive viable cells and intensity grades 0–3, then calculate an H-score from the intensity-weighted percentages (standard IHC practice). If cell counts vary markedly, report positive cells per mm² alongside the fraction positive, using the same region-selection rules across samples (standard IHC practice). Normalize comparisons to the annotated viable cell population and keep staining batches and image-analysis thresholds consistent; score nuclear and cytoplasmic signal separately (standard IHC practice; UniProt O00141: subcellular location).
How do I distinguish SGK1 staining from tissue artefact?
A credible result follows intact cells and a plausible compartment: tissue IHC shows general cytoplasmic expression, while SGK1 can also enter nuclei under specific conditions (HPA: tissue IHC profile; UniProt O00141: subcellular location). Compare the claimed positive population with a matched control and examine cell borders at higher magnification before assigning membranous signal (standard IHC practice; UniProt O00141: subcellular location). Treat staining confined to cut edges, necrotic regions, or no-primary controls as suspect, and check peroxidase blocking when unexplained DAB deposits persist (standard IHC practice). Interpret discordance cautiously because the tissue atlas rates its SGK1 staining Approved while reporting low consistency with RNA expression (HPA: reliability).
Boster reagents
Best SGK1 / Serine/threonine-protein kinase Sgk1 IHC Antibodies
Anti-SGK1 IHC images show human mammary, breast and rectal cancer, human stomach, and mouse and rat ovary; IF images show A549 and U20S cells and human stomach (catalog image captions).
Real IHC data
Anti-SGK1 Antibody ®
Cat # A00673-2
Real IHC data
Anti-SGK1/Sgk Rabbit Monoclonal Antibody
Cat # M00673
Real IHC data
Anti-SGK1 Antibody ®
Cat # A00673
Real IHC data
Anti-SGK1 Antibody
Cat # A00673-1
A00673-2 is shown on paraffin sections of human mammary cancer, while M00673 is shown on paraffin sections of human breast cancer and mouse and rat ovary (A00673-2 and M00673 IHC captions). A00673 is shown on a paraffin section of human rectal cancer, and A00673-1 is shown on human stomach tissue (A00673 and A00673-1 IHC captions).
Which to pick: For human tissue IHC, choose A00673-2 for mammary cancer at 1 μg/mL, A00673 for rectal cancer at 1 μg/mL, or A00673-1 for stomach tissue at 5 μg/mL, using each antibody’s own IHC image as the starting point (respective IHC captions). For IF/ICC, A00673-2 has an A549 cell image at 4 μg/mL and A00673 has a U20S cell image at 2 μg/mL; A00673-1 has an IF image of human stomach at 20 μg/mL (catalog applications; respective IF captions). For cross-species tissue IHC, choose rabbit monoclonal M00673, clone EHF-19: its IHC images include human breast cancer and mouse and rat ovary at 1:50 (catalog clone and reactivity; M00673 IHC captions); the fixative is unreported in the IHC captions for all four antibodies (catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00141 (SGK1_HUMAN, Serine/threonine-protein kinase Sgk1).
- Human Protein Atlas. SGK1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SGK1 subcellular location (ICC-IF): Mainly localized to the nuclear speckles..
- Human Protein Atlas. SGK1 antibody validation summary (3 antibodies).
- Induction of the cell survival kinase Sgk1: A possible novel mechanism for α-phenyl-N-tert-butyl nitrone in experimental stroke. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 2019 — PMC6545623.
- High-Salt Diet Exacerbates Murine Imiquimod-Induced Psoriasis-like Dermatitis via Activation of Serum- and Glucocorticoid-Inducible Kinase 1. International Journal of Molecular Sciences 2026 — PMC13607775.
- SGK1 Mediates Hypoxic Pulmonary Hypertension through Promoting Macrophage Infiltration and Activation. Analytical cellular pathology (Amsterdam) 2019 — PMC6877960.
- Association between Decreased SGK1 and Increased Intestinal α-Synuclein in an MPTP Mouse Model of Parkinson's Disease. International journal of molecular sciences 2023 — PMC10671719.
- PubMed PMID:9114008 — UniProt-cited evidence.
- PubMed PMID:9722955 — UniProt-cited evidence.
- PubMed PMID:18753299 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SGK1 (Serine/threonine-protein kinase Sgk1). Expected localisation Predominantly cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00673-2). Validated IHC antibody: A00673-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Soft tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human mammary cancer tissue; fixative not specified (datasheet A00673-2); 2. Fixation: Image fixative and duration unreported (datasheet A00673-2); 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 A00673-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00673-2); 8. Primary antibody: Rabbit anti-SGK1, 0.5-1μg/ml (datasheet A00673-2); 9. Primary incubation: Overnight at 4 °C (datasheet A00673-2). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Cell cycle and stimuli can shift SGK1 localisation (UniProt); Isoform / epitope: 5 isoforms; check antibody epitope coverage (UniProt); Regulation: Staining intensity varies by tissue (HPA tissue IHC). Evidence: UniProt O00141, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nuclear speckles., Datasheet A00673-2, PMC6545623, PMC13607775. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SGK1/immunohistochemistry-ihc-guide.