SASS6 / Spindle assembly abnormal protein 6 homolog · IHC design guide

Design Immunohistochemistry for SASS6

Plan paraffin-section SASS6 IHC using the IHC-validated antibody at 2–5 μg/ml (datasheet A05762-1). Compare medium staining in endometrial glandular cells with undetected staining in adrenal glandular cells, while accounting for uncertain tissue staining reliability (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for SASS6 (IHC for SASS6): expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A05762-1, validated IHC image, and IHC protocol steps — Printable **SASS6** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A05762-1, controls and protocol steps. Open the full SASS6 IHC guide →

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SASS6 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 tissue staining (HPA tissue IHC)
	Staining pattern	Cytoplasmic staining in most tissues and some immune cells (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A05762-1)
	Positive control ⓘ	Adipose tissue+4 more · see all
	Negative control ⓘ	Adrenal gland+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific)
	Caveat	Tissue staining is uncertain; external verification is pending (HPA tissue IHC)
	Regulation	Expression regulation is not annotated (UniProt)
	Isoform / epitope	No annotated isoforms or processing; one 1–657 chain (UniProt)

Section 1

Recommended SASS6 IHC & IF Protocols

Compare the catalog antibody's IHC-P protocol (datasheet A05762-1) with the published paraffin-section protocol (PMC11499408).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human appendiceal carcinoid tissue; fixative not specified (datasheet A05762-1)
Fixation	Image fixative and duration unreported (datasheet A05762-1); 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 A05762-1); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A05762-1)
Primary antibody	Rabbit anti-SASS6, 2-5 μg/ml (datasheet A05762-1)
Primary incubation	Overnight at 4 °C (datasheet A05762-1)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A05762-1)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	SASS6-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in most tissues and in a subset of immune cells. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A05762-1); the published protocol does not specify retrieval (PMC11499408).

Section 2

What Is the Expected SASS6 Staining Pattern?

SASS6 is associated with centrosomes and nascent procentrioles, with no transmembrane segment (UniProt Q6UVJ0). In paraffin section IHC, assess cytoplasmic staining in the cell populations reported by HPA, such as endometrial glandular cells or kidney glomerular cells, both stained at medium intensity (HPA: tissue IHC). Treat that pattern as provisional: HPA rates the tissue staining and its listed antibody HPA028187 as uncertain (HPA: tissue IHC; HPA: antibody validation).

What am I looking at on my slide?
Cytoplasmic staining in endometrial glandular cells or kidney glomerular cells, with limited staining in adjacent cells.	This is compatible with HPA's medium staining observations in those cell populations (HPA: tissue IHC). Score the relevant cells and compartment; the uncertain tissue and antibody ratings mean the pattern alone does not establish specificity (HPA: tissue IHC; HPA: antibody validation).
Small cytoplasmic foci are visible, but many otherwise stained cells lack a resolvable punctum.	A focal component is plausible because SASS6 associates transiently with nascent procentrioles (UniProt Q6UVJ0). Chromogenic section IHC may not resolve every centriole; interpret the overall cellular pattern alongside controls rather than requiring a punctum in each positive cell (standard IHC practice).
Predominantly nuclear or membrane rim staining appears without a convincing cytoplasmic component.	That compartment pattern conflicts with UniProt's centrosome and centriole annotation and its lack of a transmembrane segment (UniProt Q6UVJ0). Consider nonspecific staining or a detection artefact, and check controls before assigning it to SASS6 (standard IHC practice).
Strong staining appears in alveolar cells or cardiomyocytes while the selected comparison tissue remains faint.	HPA reports SASS6 as not detected in those respective cell populations (HPA: lung alveolar cells; HPA: heart cardiomyocytes). This mismatch raises concern for cross-reactivity or endogenous detection activity; it is a warning, not proof, given HPA's uncertain IHC reliability (HPA: tissue IHC).
The whole section has a diffuse tint, or no cells stain in a selected HPA medium-staining tissue.	Diffuse colour obscures compartment scoring; a blank comparison tissue makes this run uninformative (standard IHC practice). Repeat with suitable detection controls, and remember that HPA's medium observations are provisional rather than guaranteed positives (HPA: tissue IHC, uncertain).

💡Expected SASS6 appearanceCall a result compatible when cytoplasmic staining in an HPA medium-staining cell population is distinguishable from background; isolated strong nuclear, membrane rim, or pan-tissue colour is suspect (HPA: tissue IHC, uncertain; UniProt Q6UVJ0; standard IHC practice).

How each factor affects the staining
Cell population used for interpretation	HPA reports medium staining in adipocytes, caudate glia, selected endothelial and glandular cells, and kidney glomerular cells (HPA: tissue IHC). It also reports low or undetected staining in specified other populations; score the annotated cell type, not a whole-organ average (HPA: tissue IHC).
Intracellular location and topology	SASS6 is a centrosome and centriole component associated transiently with nascent procentrioles; it has no transmembrane segment (UniProt Q6UVJ0). Cytoplasmic staining can be assessed in section IHC, while a sharply resolved centriole is not required for a compatible chromogenic result (standard IHC practice).
Strength of IHC evidence	HPA describes cytoplasmic expression in most tissues and a subset of immune cells, but labels tissue staining uncertain and HPA028187 IHC uncertain (HPA: tissue IHC; HPA: antibody validation). Use tissue comparisons and controls to qualify an interpretation; HPA's reported intensity is not a validated threshold.
IF/ICC Q: Should the IHC result reproduce ciliary IF staining?	No direct match is established. HPA approves primary cilium localization in ICC-IF but marks cytosol and vesicles uncertain (HPA: subcellular). UniProt places SASS6 at centrosomes and centrioles (UniProt Q6UVJ0). Interpret each application using its own imaging context.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
No staining in the selected endometrial glandular or kidney glomerular cells.	The run may have failed, or the selected section may differ from HPA's medium-staining observation; HPA rates its IHC evidence uncertain (HPA: tissue IHC).	Confirm that the intended cells are present, inspect the run controls, and review the catalog antibody's IHC-P instructions before repeating (standard IHC practice). Do not infer absence of SASS6 from a single blank section.
Diffuse brown colour makes cytoplasmic staining hard to distinguish.	Nonspecific background or detection activity can obscure cellular localization (standard chromogenic IHC practice).	Compare with a no-primary control; review blocking, washing, and detection exposure. If using peroxidase detection, check the endogenous enzyme quench (standard IHC practice). Reassess only after background permits compartment scoring.
Alveolar cells or cardiomyocytes stain strongly.	Those cells are reported as not detected by HPA, so cross-reactivity or endogenous detection activity is possible (HPA: lung alveolar cells; HPA: heart cardiomyocytes).	Compare the same run with an HPA medium-staining cell population and a no-primary control; inspect whether colour tracks cellular compartments or tissue-wide background (HPA: tissue IHC; standard IHC practice).
Signal is mainly nuclear or outlines cell membranes.	That distribution conflicts with the annotated centrosome and centriole location and absence of a transmembrane segment (UniProt Q6UVJ0).	Check the no-primary control and tissue morphology, then reassess antibody specificity with an independent control if available (standard IHC practice). Do not score the misplaced signal as a confident SASS6 positive.
Only weak cytoplasmic staining is visible in a tissue expected to stain.	HPA reports low staining in some cell populations and medium staining in others; its overall IHC reliability is uncertain (HPA: tissue IHC). The sampled cells may affect the comparison.	Identify and score the exact cell population, compare sections processed in the same run, and confirm that the signal exceeds background (standard IHC practice). Avoid applying a single intensity cutoff across tissues.
A ciliary IF image seems inconsistent with the section IHC pattern.	HPA's approved primary cilium location comes from ICC-IF, while its tissue IHC describes cytoplasmic expression and is uncertain (HPA: subcellular; HPA: tissue IHC).	Keep the applications' observations distinct: score the cell type and chromogenic compartment on the section, and consult the separate IF/ICC guide for fluorescence interpretation (standard IHC/IF practice).

Source: UniProt Q6UVJ0 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing SASS6 staining

IHC analysis of SASS6 using anti-SASS6 antibody (A05762-1). SASS6 was detected in a paraffin-embedded section of human appendiceal carcinoid 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-SASS6 Antibody (A05762-1) 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.

A05762-1 · IHCIHC analysis of SASS6 using anti-SASS6 antibody (A05762-1). SASS6 was detected in a paraffin-embedded section of human appendiceal carcinoid

IF analysis of SASS6 using anti-SASS6 antibody (A05762-1). SASS6 was detected in a paraffin-embedded section of human testis 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 5 μg/mL rabbit anti-SASS6 Antibody (A05762-1) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A05762-1 · IFIF analysis of SASS6 using anti-SASS6 antibody (A05762-1). SASS6 was detected in a paraffin-embedded section of human testis cancer tissue.

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000156876-SASS6/tissue/cerebral+cortex#imid_7136688

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000156876-SASS6/tissue/colon#imid_7136630

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000156876-SASS6/tissue/liver#imid_7136631

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000156876-SASS6/tissue/kidney#imid_7136624

HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA028187; green = SASS6, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for SASS6 IHC & IF

🧪Run adipose tissue first and score staining in adipocytes, where SASS6 is reported at a medium level (HPA: adipose tissue, adipocytes, Medium). Use adrenal gland glandular cells as a negative tissue (HPA: adrenal gland, glandular cells, Not detected); on the adipose slide, compare adipocytes with adjacent non-adipocyte cells, but do not presume those cells are SASS6-negative because their staining is not specified in the supplied HPA row.

Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)

Negative control tissue: Adrenal gland (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SASS6 in U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Primary cilium (approved), Cytosol (uncertain) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control, a clonality-matched nonimmune rabbit IgG isotype control, and SASS6-knockout tissue or cells as a biological negative (selected-SKU caption: rabbit primary antibody). For chromogenic staining, block endogenous peroxidase and check signal in blood or inflammatory cells within the adipose section (selected-SKU caption: HRP/DAB detection; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but provides no comparison that establishes retrieval dependence (selected-SKU caption: EDTA retrieval); the supplied evidence does not establish whether frozen sections or IF would be easier. Adipose sections can show lipid-extraction spaces and tissue tearing, so assess morphology before scoring chromogenic staining (standard IHC practice).

HPA tissue IHC evidence for SASS6

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Antibody staining in cells/structures not annotated, view images. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Medium	Protein (IHC)	HPA →
Breast	Adipocytes	Medium	Protein (IHC)	HPA →
Caudate	Glial cells	Medium	Protein (IHC)	HPA →
Cerebral cortex	Endothelial cells	Medium	Protein (IHC)	HPA →
Colon	Endothelial cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adrenal gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Appendix	Glandular cells	Not detected	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	Not detected	Protein (IHC)	HPA →
Fallopian tube	Glandular cells	Not detected	Protein (IHC)	HPA →
Heart muscle	Cardiomyocytes	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SASS6 IHC Tips

Troubleshoot chromogenic SASS6 IHC in paraffin sections using the catalog antibody’s tissue image, UniProt localisation, and cautiously interpreted HPA observations.

Which retrieval conditions should I try first for SASS6 in paraffin sections?

Start with heat-mediated retrieval in EDTA at pH 8.0 for this catalog antibody (datasheet A05762-1). The product image used this retrieval before staining a paraffin section of human appendiceal carcinoid tissue, so keep the antibody and detection conditions comparable during the first run (datasheet A05762-1). If staining is weak, vary heating duration on matched sections while checking morphology; treat another buffer as a fallback to test rather than an established SASS6 condition (standard IHC practice). Include a no-primary control and compare signal in intact cells, because excessive retrieval can make background and tissue damage harder to distinguish from staining (standard IHC practice).

How should I assess fixation as a cause of weak SASS6 staining?

The selected tissue caption identifies a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A05762-1). Record fixative, fixation duration, processing history, and section age for each specimen, then compare sections processed together before changing antibody concentration (standard IHC practice). Examine whether weak staining follows poorly preserved regions or entire processing batches, using morphology and a suitable tissue control to separate preservation problems from staining variability (standard IHC practice). Avoid assigning a SASS6-specific fixation effect to any pattern without a matched comparison, and retain a no-primary section to assess detection background (standard IHC practice).

Should SASS6 appear as diffuse cytoplasmic staining or discrete puncta?

SASS6 is associated with centrosomes and transiently with nascent procentrioles during centriole biogenesis (UniProt Q6UVJ0). Its small structures may be difficult to resolve with chromogenic light microscopy, so record punctate and broader cytoplasmic staining separately rather than requiring every positive cell to show a dot (UniProt Q6UVJ0; standard IHC practice). HPA describes cytoplasmic staining in most tissues, but rates its tissue IHC reliability as Uncertain, which limits its value as a definitive localisation standard (HPA tissue IHC). Treat dominant nuclear or extracellular DAB as a reason to inspect morphology, background controls, and antibody specificity before assigning it to SASS6 (UniProt Q6UVJ0; standard IHC practice).

Could isoforms or epitope location explain inconsistent SASS6 IHC?

The supplied record lists 0 isoforms and a single 1–657 chain, so an isoform explanation lacks support here (UniProt Q6UVJ0). SASS6 has no annotated transmembrane segment or signal peptide, placing the expected antigen in an intracellular compartment for access during section staining (UniProt Q6UVJ0). Its PISA domain spans residues 39–91, and annotated phosphoserines include 510, 615, and 657; these annotations do not identify this antibody’s epitope (UniProt Q6UVJ0). If batches differ, confirm the antibody’s documented immunogen or epitope before attributing staining changes to phosphorylation, and compare matched processing controls (standard IHC practice).

How can IF help assess an ambiguous SASS6 IHC pattern?

Use IF as a complementary localisation check, since SASS6 is associated with centrosomes and nascent procentrioles while HPA also reports primary-cilium localisation (UniProt Q6UVJ0; HPA subcellular). Multiplex with a validated marker for the expected cell type and, when testing puncta, a separately labelled centrosome or cilium marker; assess channel alignment and single-stain controls (standard IF practice). Choose fluorophores after measuring the tissue’s autofluorescence, favouring a spectrally separated channel with adequate signal for the SASS6 antibody (standard IF practice). Because SASS6 lacks a transmembrane segment, test permeabilisation for intracellular epitope access and verify that the treatment preserves punctate structures (UniProt Q6UVJ0; standard IF practice).

What should I change when DAB background obscures SASS6 staining?

First compare the stained section with a no-primary control to locate background from secondary detection, endogenous peroxidase, or nonspecific chromogen deposition (standard IHC practice). The selected image used 10% goat serum block, 2 μg/ml primary antibody overnight at 4°C, and DAB detection, providing defined starting conditions for this antibody (datasheet A05762-1). Apply an appropriate peroxidase block, check washing and reagent carryover, then titrate the primary on matched sections if diffuse colour remains (standard IHC practice). Evaluate edges, folds, and damaged areas separately; their staining should not establish SASS6 positivity without intact-cell signal and clean controls (standard IHC practice).

How should I score SASS6 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, then report the percentage of positive intact cells and an intensity-based H-score if diffuse cytoplasmic DAB is reproducible (standard IHC practice). For discrete puncta, count positive cells or puncta per mm² of evaluable tissue and document the optical criteria used to distinguish dots from pigment (standard IHC practice). Normalise comparisons to the same cell population, evaluable area, staining batch, and exposure or scanning settings, while excluding folds and necrosis (standard IHC practice). Keep cytoplasmic and punctate measures separate because SASS6 is centrosome associated, whereas HPA’s broader tissue staining carries Uncertain reliability (UniProt Q6UVJ0; HPA tissue IHC).

When is a SASS6-positive DAB signal convincing?

Give greatest weight to reproducible staining in intact cells with plausible intracellular or centrosome-associated distribution, clean no-primary controls, and consistent morphology across sections (UniProt Q6UVJ0; standard IHC practice). Check the cell identity: HPA reports medium staining in several cell populations, including adipocytes and glial, endothelial, and glandular cells, but rates its tissue IHC observations Uncertain (HPA tissue IHC). Do not equate the catalog image of appendiceal carcinoid tissue with normal appendix glandular cells, which HPA lists as not detected (datasheet A05762-1; HPA tissue IHC). Discount isolated nuclear or extracellular colour, section-edge staining, necrotic deposits, and signal retained in peroxidase or no-primary controls before calling SASS6 positive (UniProt Q6UVJ0; standard IHC practice).

Boster reagents

Best SASS6 / Spindle assembly abnormal protein 6 homolog IHC Antibodies

A05762-1 has IHC data from human paraffin sections of appendiceal carcinoid and breast cancer, plus IF data from a human paraffin section of testis cancer (catalog image captions).

Real IHC data

Anti-SASS6 Antibody ®

Cat # A05762-1

View product →

A05762-1 has IHC images from human paraffin sections of appendiceal carcinoid and breast cancer (catalog IHC captions). Its IF image shows a human paraffin section of testis cancer; the catalog lists Human, Mouse and Rat reactivity (catalog IF caption; catalog reactivity).

Which to pick: Choose A05762-1 for tissue IHC: its IHC captions document staining of human paraffin sections with the rabbit antibody, while the fixative is unreported (catalog IHC captions). For IF, A05762-1 has a human tissue section image, but the supplied data do not establish ICC validation (catalog IF caption; catalog applications). For work across species, A05762-1 lists Human, Mouse and Rat reactivity; its IHC dilution listing specifies Human and Mouse, and the supplied IHC images show human tissue (catalog reactivity; catalog dilution listing; catalog IHC captions).

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

References

UniProt Consortium. UniProt entry Q6UVJ0 (SAS6_HUMAN, Spindle assembly abnormal protein 6 homolog).
Human Protein Atlas. SASS6 tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. SASS6 subcellular location (ICC-IF): Mainly localized to the cytosol and primary cilium. In addition localized to vesicles..
Human Protein Atlas. SASS6 antibody validation summary (1 antibodies).
SASS6 promotes tumor proliferation and is associated with TP53 and immune infiltration in lung adenocarcinoma. Clinical and experimental medicine 2024 — PMC11499408.
PubMed PMID:16710414 — UniProt-cited evidence.
PubMed PMID:17974005 — UniProt-cited evidence.
PubMed PMID:15572125 — UniProt-cited evidence.