This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SLAIN2 / SLAIN motif-containing protein 2 · IHC design guide
Design Immunohistochemistry for SLAIN2
Plan SLAIN2 IHC in paraffin sections using cytoplasmic staining as the expected tissue pattern (HPA tissue IHC). Compare high-staining myocytes and smooth muscle cells with an unstained cell population, and optimize the catalog antibody from its stated 5 μg/mL starting concentration (HPA tissue IHC; datasheet: 5 μg/mL).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SLAIN2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining, high in myocytes and smooth muscle cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Skeletal muscle+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 | Staining varies by cell type; adipocytes were unstained (HPA tissue IHC) | |
| Regulation | Stimulus-linked regulation is unreported (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–581 chain, with no transmembrane segment (UniProt) |
Section 1
Recommended SLAIN2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is accompanied by a published SLAIN2 IHC workflow in colorectal cancer tissue (PMC6444028: IHC methods).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A11030); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-SLAIN2, 5 μg/mL (datasheet A11030) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SLAIN2-positive staining in myocytes of skeletal muscle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues, high expression in testis and smooth muscle. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); compare EDTA retrieval (PMC6444028: IHC methods).
Section 2
What Is the Expected SLAIN2 Staining Pattern?
SLAIN2 localizes to the cytoplasm and microtubule cytoskeleton and has no transmembrane segment (UniProt Q9P270: localization/topology). In tissue IHC, expect cytoplasmic staining in many tissues, with High staining in skeletal myocytes and smooth muscle cells among the listed examples (HPA tissue IHC). HPA rates the tissue staining Approved, with external verification pending (HPA tissue IHC: reliability).
| Strong cytoplasmic staining in skeletal myocytes or smooth muscle cells, with recognizable cell boundaries (HPA tissue IHC: High). | This matches the reported cell types and compartment (HPA tissue IHC: High; UniProt Q9P270: cytoplasm). Judge intensity against nearby tissue structures and the staining control; a brown deposit alone does not establish cell identity (standard IHC practice). |
| Predominantly nuclear staining with little cytoplasmic signal in otherwise intact cells. | A nuclear-only pattern conflicts with the reported cytoplasmic localization (UniProt Q9P270; HPA tissue IHC). Check morphology, counterstain and detection controls before assigning it to SLAIN2; the supplied sources do not establish a nuclear tissue-IHC pattern. |
| Strong staining in adipocytes from adipose tissue or breast, or in another cell type listed as not detected (HPA tissue IHC). | Treat this as unexpected for that specified cell type, with cross-reactivity or endogenous chromogen activity among the possibilities (HPA tissue IHC; standard IHC practice). HPA's not-detected entries describe named cells, not every cell in the tissue. |
| Diffuse brown haze across cells and extracellular spaces, without a discernible cytoplasmic pattern. | This is difficult to score as specific SLAIN2 staining because the expected tissue pattern is cellular and cytoplasmic (HPA tissue IHC). Background from detection reagents or insufficient blocking is possible; compare a no-primary control (standard IHC practice). |
| No staining in skeletal myocytes or smooth muscle cells that are present and well preserved on the section (HPA tissue IHC: High). | This fails the supplied positive-tissue expectation, but one blank section cannot identify the cause (HPA tissue IHC). Check tissue identity, antibody and detection controls, and the documented IHC-P conditions before concluding that the sample lacks SLAIN2 (standard IHC practice). |
💡Expected SLAIN2 appearanceA convincing positive is cytoplasmic staining in identified skeletal myocytes or smooth muscle cells, potentially High in intensity (HPA tissue IHC; UniProt Q9P270); isolated nuclear staining or stain confined to a cell type listed as not detected warrants investigation (HPA tissue IHC; UniProt Q9P270).
| Compartment and topology | UniProt places SLAIN2 in the cytoplasm and microtubule cytoskeleton, including growing microtubule plus ends, and reports no transmembrane segment (UniProt Q9P270). Interpret tissue IHC primarily at the cytoplasmic level; the record does not require visible plus-end puncta in chromogenic sections. |
| Choice of tissue and cell type | HPA reports High staining in skeletal myocytes and smooth muscle cells, Medium in several other listed cell types, and not-detected staining in specified cells such as adipocytes (HPA tissue IHC). Select and score controls by cell type; whole-tissue labels can conceal mixed cell populations. |
| Strength of antibody evidence | The tissue-IHC profile is Approved but pending external verification (HPA tissue IHC: reliability). HPA063599 is IHC Approved; HPA035824 has no IHC status listed (HPA antibodies). A missing status is not evidence of failed staining, and the supplied record does not establish two-antibody IHC confirmation. |
| IF/ICC Q: What localization may appear? | A: HPA reports supported cytosol and centrosome localization, plus primary cilium tip and basal body localization, in ICC-IF (HPA subcellular). Those finer structures are IF observations; their visibility is not a requirement for a positive chromogenic tissue-IHC section. |
| Chromogenic background | Endogenous detection activity and nonspecific reagent binding can create signal unrelated to the primary antibody (standard IHC practice). Use an appropriate no-primary or detection control to help distinguish such signal from the HPA-reported cytoplasmic cell pattern (HPA tissue IHC; standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known positive muscle cells are blank (HPA tissue IHC: High). | Tissue identification, staining execution, or antibody and detection performance may be responsible; the supplied sources provide no SLAIN2-specific fixation effect (standard IHC practice). | Confirm the muscle cell type and section quality, then check controls and the catalog antibody's documented IHC-P retrieval and dilution conditions before changing one workflow variable at a time (standard IHC practice). |
| Nuclei dominate while cytoplasm is weak. | This conflicts with cytoplasmic localization and may reflect background, counterstain interpretation, or nonspecific binding (UniProt Q9P270; standard IHC practice). | Inspect the no-primary control and cell morphology, then reassess blocking and detection conditions; withhold a positive SLAIN2 call until cytoplasmic staining is distinguishable (standard IHC practice). |
| Adipocytes stain strongly in an HPA not-detected context (HPA tissue IHC). | Unexpected cell staining may arise from cross-reactivity or endogenous detection activity; the HPA result applies to the named cell population (HPA tissue IHC; standard IHC practice). | Verify cell identity on the counterstained section, compare an appropriate detection control, and assess staining in listed positive muscle cells on the same run (HPA tissue IHC; standard IHC practice). |
| Brown haze obscures cell boundaries. | Nonspecific binding or background from the chromogenic detection system can prevent reliable compartment scoring (standard IHC practice). | Compare a no-primary control, check blocking and washes, and adjust the antibody or detection conditions within the documented IHC-P workflow; rescore only when cytoplasm can be distinguished (standard IHC practice). |
| The IHC section lacks centrosomal or ciliary puncta seen in IF/ICC. | HPA's supported centrosome, cilium-tip and basal-body localizations come from ICC-IF; the tissue-IHC profile describes cytoplasmic expression (HPA subcellular; HPA tissue IHC). | Score tissue IHC using cell identity and cytoplasmic signal. Do not require IF-resolved puncta as an IHC acceptance criterion (HPA tissue IHC; HPA subcellular). |
Experiment evidence
IHC & IF images showing SLAIN2 staining
A11030 · IHCImmunohistochemistry of SLAIN2 in human testis with SLAIN2 antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109171-SLAIN2/tissue/cerebral+cortex#imid_20060557
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109171-SLAIN2/tissue/colon#imid_20060483
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109171-SLAIN2/tissue/liver#imid_20060500
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109171-SLAIN2/tissue/kidney#imid_20060493
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA035824; green = SLAIN2, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA035824; green = SLAIN2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SLAIN2 IHC & IF
🧪Run skeletal muscle first: myocytes should stain (HPA: High in skeletal muscle myocytes). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); on the positive slide, treat staining outside myocytes as background only where it matches the no-primary control, rather than assuming every other cell type is negative.
Positive control tissue: Skeletal muscle (Myocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SLAIN2 in U-251MG, U2OS, Rh30, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Centrosome (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide, an isotype control matched to the primary antibody’s host species and clonality, and a SLAIN2 knockout sample or validated peptide-block control (standard IHC controls). For skeletal muscle chromogenic IHC, quench endogenous peroxidase and check the no-primary slide for residual tissue pigment or background (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize retrieval empirically for paraffin sections. The selected A11030 testis IHC caption reports 5 μg/mL but leaves the fixative unreported (selected-SKU tissue-IHC caption). HPA reports supported cytosol and centrosome localization by ICC-IF, but the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC (HPA: subcellular localization); assess muscle background with the no-primary slide (standard IHC practice).
HPA tissue IHC evidence for SLAIN2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SLAIN2 IHC Tips
Troubleshoot SLAIN2 staining in paraffin section IHC by checking retrieval, controls and the expected cytoplasmic pattern before scoring (UniProt Q9P270; HPA tissue IHC).
What retrieval should I try first if SLAIN2 staining is weak in paraffin sections?
Start with citrate pH 6.0 heat induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule). Allow sections to cool in the retrieval buffer, then keep the heating and cooling conditions consistent across control and study slides (standard IHC practice). Use a human testis control when evaluating the catalog antibody: its selected tissue IHC caption shows staining at 5 μg/mL, but reports no retrieval method (catalog antibody A11030 caption). If staining remains weak, compare a more alkaline retrieval buffer on matched sections as a fallback, checking whether any gain also raises background (standard IHC practice). Cytoplasmic signal is the expected readout (HPA tissue IHC).
How should I troubleshoot variable SLAIN2 staining between differently fixed samples?
SLAIN2 specific sensitivity to fixation is unknown from the supplied evidence; the selected human testis caption gives 5 μg/mL but does not state a fixative (catalog antibody A11030 caption). Record fixative, fixation duration, tissue thickness and processing history for each block before comparing staining intensity (standard IHC practice). Run matched sections through the same citrate pH 6.0, 95–98 °C, 20 min retrieval and detection sequence to isolate processing differences (page retrieval rule; standard IHC practice). Include a concurrently processed positive control and assess staining by cell compartment, since SLAIN2 is associated with the cytoplasm and cytoskeleton (UniProt Q9P270). Do not assign a fixation effect from tissue staining differences alone (standard IHC practice).
Which compartment should count as convincing SLAIN2 staining in chromogenic IHC?
Prioritize cytoplasmic staining within identifiable cells: SLAIN2 is assigned to the cytoplasm and cytoskeleton, with association at growing microtubule plus ends (UniProt Q9P270). HPA likewise reports cytoplasmic expression in most tissues and supported cytosol and centrosome localisation (HPA tissue IHC; HPA subcellular). A diffuse nuclear only pattern should prompt review of the counterstain, detection background and primary antibody control before it is scored as SLAIN2 (standard IHC practice; UniProt Q9P270). Fine centrosomal or microtubule associated detail may be difficult to resolve with chromogenic tissue IHC, so score the cellular compartment you can actually distinguish (standard IHC practice). Compare the same cell type across sections (standard IHC practice).
Could an isoform or hidden epitope explain inconsistent SLAIN2 IHC?
The supplied record lists 0 isoforms and one annotated chain spanning residues 1–581, so an isoform specific explanation is unsupported here (UniProt Q9P270). It lists no transmembrane segment or signal peptide, and places SLAIN2 in the cytoplasm and cytoskeleton (UniProt Q9P270). The antibody epitope is not supplied, so neither its residue range nor its fixation sensitivity can be inferred from those annotations (supplied record; catalog antibody A11030 caption). SLAIN2 has annotated modified residues, including phosphoserines at 43, 48, 63 and 88, but their effect on this antibody is unknown (UniProt Q9P270). Check the antibody’s mapped immunogen or epitope before attributing inconsistent staining to modification (standard IHC practice).
How can IF help assess SLAIN2 staining when multiplexing tissue markers?
Use IF as a companion localisation check while keeping chromogenic paraffin section IHC as the scoring assay (standard IHC/IF practice). Pair SLAIN2 with a marker for the expected cell type, such as a smooth muscle cell marker when assessing smooth muscle, where HPA reports high staining (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and favor a far red SLAIN2 channel if tissue autofluorescence obscures shorter wavelength channels (standard IF practice). SLAIN2 has no annotated transmembrane segment and is cytoplasmic, so assess whether gentle permeabilisation exposes the intracellular epitope; its exact epitope is unreported (UniProt Q9P270; supplied record; standard IF practice). Include single stain and no primary controls when judging overlap (standard IF practice).
What should I check when SLAIN2 IHC shows widespread brown background?
First compare the suspect slide with a no primary control and inspect whether deposits track tissue edges, folds or damaged regions (standard IHC practice). For chromogenic detection, verify the peroxidase block and check for nonspecific secondary reagent or DAB deposition; these are general workflow checks, not demonstrated SLAIN2 effects (standard IHC practice). Titrate around the catalog antibody’s reported 5 μg/mL testis image concentration using matched sections, because that caption does not establish an optimal concentration for every tissue (catalog antibody A11030 caption; standard IHC practice). Judge signal in identifiable cytoplasm, the expected broad compartment, against the control rather than calling every brown pixel positive (HPA tissue IHC; standard IHC practice). Keep development time identical across slides (standard IHC practice).
How should I quantify SLAIN2 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring rule before reading the slides, because HPA reports cell dependent staining across tissues (HPA tissue IHC; standard IHC practice). For comparable regions, record percent positive cells and an intensity based H-score, or positive cell density per mm² when cell counts matter (standard IHC practice). Normalize to the number of evaluable cells or measured tissue area, and exclude folds, necrosis and section edges using the same criteria throughout (standard IHC practice). Keep retrieval at citrate pH 6.0, 95–98 °C for 20 min and detection conditions consistent within a comparison (page retrieval rule; standard IHC practice). Report scores by cell type rather than averaging unlike compartments (standard IHC practice).
How do I distinguish true SLAIN2 staining from tissue artefact?
A credible positive is intracellular and predominantly cytoplasmic in an identifiable cell, matching the reported SLAIN2 compartment (UniProt Q9P270; HPA tissue IHC). HPA reports high staining in skeletal muscle myocytes and smooth muscle cells, whereas adipocytes in adipose tissue were not detected; use those observations as context rather than absolute pass or fail criteria (HPA tissue IHC). Review nuclear only staining, staining confined to edges or necrotic areas, and signal reproduced in the no primary control as possible artefact (standard IHC practice; UniProt Q9P270). Check peroxidase associated colour against the blocked control before attributing brown deposits to SLAIN2 (standard IHC practice). HPA labels its tissue staining Approved and pending external verification, so avoid treating one pattern as definitive (HPA tissue IHC).
Boster reagents
Best SLAIN2 / SLAIN motif-containing protein 2 IHC Antibodies
The IHC-validated anti-SLAIN2 antibody A11030 has real IHC data from human testis (catalog image caption); its listed application is IHC-P (catalog applications).
Real IHC data
Anti-SLAIN2 Antibody
Cat # A11030
A11030 is listed for human IHC-P (catalog applications; catalog reactivity). Its IHC image shows human testis stained at 5 μg/mL (catalog image caption).
Which to pick: Choose A11030 for paraffin-section tissue IHC: it is a rabbit polyclonal antibody listed for human IHC-P, with an image from human testis (catalog host; catalog dilution description; catalog applications; catalog image caption). The fixative used for that image is unreported (catalog image caption). No SKU in the payload is listed for IF/ICC or species beyond human (catalog applications; catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9P270 (SLAI2_HUMAN, SLAIN motif-containing protein 2).
- Human Protein Atlas. SLAIN2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SLAIN2 subcellular location (ICC-IF): Mainly localized to the cytosol and centrosome. In addition localized to the primary cilium tip and basal body..
- Human Protein Atlas. SLAIN2 antibody validation summary (2 antibodies).
- MALAT1 sponges miR-106b-5p to promote the invasion and metastasis of colorectal cancer via SLAIN2 enhanced microtubules mobility. EBioMedicine 2019 — PMC6444028.
- PubMed PMID:10819331 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SLAIN2 (SLAIN motif-containing protein 2). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A11030 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Skeletal muscle (Myocytes); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A11030); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-SLAIN2, 5 μg/mL (datasheet A11030); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining varies by cell type; adipocytes were unstained (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–581 chain, with no transmembrane segment (UniProt); Regulation: Stimulus-linked regulation is unreported (UniProt). Evidence: UniProt Q9P270, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol and centrosome. In addition…, Datasheet A11030, PMC6444028. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SLAIN2/immunohistochemistry-ihc-guide.