TLN2 / Talin-2 · IHC design guide

Design Immunohistochemistry for TLN2

Plan chromogenic IHC on paraffin sections for TLN2 using its reported cytoplasmic tissue pattern (HPA tissue IHC). The guide covers tissue selection, staining controls and interpretation, with the reported medium consistency between antibody staining and RNA expression in mind (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for TLN2 (IHC for TLN2): expected localisation Cytoplasmic staining in positive tissue cells (HPA tissue IHC), antibody PB9961, validated IHC image, and IHC protocol steps — Printable **TLN2** IHC protocol sheet — expected localisation Cytoplasmic staining in positive tissue cells (HPA tissue IHC), antibody PB9961, controls and protocol steps. Open the full TLN2 IHC guide →

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TLN2 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 positive tissue cells (HPA tissue IHC)
	Staining pattern	Cytoplasm in neurons, cardiomyocytes and select renal tubules (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet PB9961)
	Positive control ⓘ	Caudate+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). Selected-image fixative and duration unreported (datasheet PB9961); verify before use.
	Caveat	Staining–RNA consistency is medium; verify with controls (HPA tissue IHC)
	Regulation	No expression regulation annotated (UniProt)
	Isoform / epitope	No isoforms; one chain with no extracellular domain (UniProt)

Section 1

Recommended TLN2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is paired with one published TLN2 protocol for paraffin-embedded clear cell renal cell carcinoma tissue (PMC8824820).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded mouse intestine tissue; fixative not specified (datasheet PB9961)
Fixation	Image fixative and duration unreported (datasheet PB9961); 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 PB9961); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PB9961)
Primary antibody	Rabbit anti-TLN2, 0.5-1μg/ml (datasheet PB9961)
Primary incubation	Overnight at 4 °C (datasheet PB9961)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9961)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	TLN2-positive staining in neuronal cells of caudate (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in neuronal cells, cardiac myocytes, and in subset of renal tubules and cells in the seminiferous ducts. No signal in the no-primary control.

💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet PB9961). The published microwave retrieval does not specify a buffer (PMC8824820).

Section 2

What Is the Expected TLN2 Staining Pattern?

TLN2 is a cytoplasmic protein associated with focal adhesions and the cytoskeleton; it has no transmembrane segment (UniProt Q9Y4G6). In tissue IHC, expect staining in neuronal cells, cardiomyocytes, selected renal tubules and cells in seminiferous ducts (HPA tissue IHC). HPA rates its tissue staining “Enhanced,” with medium agreement between staining and RNA data; external verification is pending (HPA tissue IHC).

What am I looking at on my slide?
Cytoplasmic staining in cardiomyocytes or neuronal cells, or along proximal-tubule microvilli.	These are reported positive sites, generally at medium intensity (HPA tissue IHC). Score the named cell population and compartment; surrounding tissue need not stain uniformly (HPA tissue IHC).
Predominantly nuclear staining, with little staining in the expected cytoplasmic sites.	A nuclear-dominant pattern conflicts with TLN2’s reported localization (UniProt Q9Y4G6; HPA subcellular). Check the counterstain and detection controls before interpreting it as TLN2 signal (general IHC practice).
Strong staining in adipocytes or bone-marrow hematopoietic cells.	HPA reports TLN2 as not detected in these cell populations (HPA tissue IHC). Treat the result as suspect and assess cross-reactivity or endogenous detection activity; an HPA-negative result is not proof that every specimen must be negative (general IHC practice).
Diffuse color across cells and extracellular areas, without cell-specific structure.	This lacks the reported cytoplasmic and cell-specific pattern (HPA tissue IHC). Review background and detection controls before scoring; diffuse color alone does not establish TLN2 expression (general IHC practice).
No staining in cardiomyocytes or neuronal cells on an otherwise interpretable section.	Those populations are reported positive at medium intensity (HPA tissue IHC). A negative slide may reflect assay performance or specimen variation; check controls before concluding that TLN2 is absent (general IHC practice).

💡Expected TLN2 appearanceCall a positive result when medium cytoplasmic staining appears in reported cells, such as cardiomyocytes or neurons (HPA tissue IHC); nuclear-dominant or indiscriminate staining is inconsistent with the reported pattern (UniProt Q9Y4G6; HPA tissue IHC).

How each factor affects the staining
Cell compartment and structure	TLN2 links integrins to the actin cytoskeleton at focal adhesions (UniProt Q9Y4G6). HPA tissue IHC describes cytoplasmic staining, including proximal-tubule microvilli; neither source predicts a nuclear-dominant IHC pattern.
Choice of tissue and cells for interpretation	HPA reports medium staining in neurons, cardiomyocytes, proximal-tubule microvilli and spermatogonia, while several other specified cell populations are not detected (HPA tissue IHC). Compare named cells rather than judging whole-section color.
Strength of tissue evidence	HPA labels two antibodies IHC “Enhanced” (HPA antibodies), but its tissue profile has medium staining–RNA consistency and awaits external verification (HPA tissue IHC). Use the reported pattern as a guide, with an assay control.
Protein architecture and epitope limits	UniProt lists one TLN2 chain, no signal peptide, no propeptide and no transmembrane segment (UniProt Q9Y4G6). The payload gives no antibody epitope or target-specific retrieval response, so these annotations cannot specify retrieval conditions.
IF/ICC Q&A: What localization can serve as a separate cross-check?	HPA supports cytosol and focal-adhesion-site localization in ICC-IF (HPA subcellular). This can inform localization interpretation, but it does not establish an IF protocol or replace the tissue IHC pattern.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
A reported positive tissue shows no convincing stain.	An uninformative assay or specimen could obscure the reported medium signal (HPA tissue IHC; general IHC practice).	Confirm that a positive control stained, then review antibody and chromogenic detection steps. Reassess the named cell population before calling the specimen negative (general IHC practice).
Color is concentrated in nuclei.	Nuclear-dominant signal conflicts with reported cytoplasmic and focal-adhesion localization (UniProt Q9Y4G6; HPA subcellular).	Check counterstain interpretation, a no-primary control and detection background; score TLN2 only where the cell and compartment pattern is credible (general IHC practice).
Adipocytes or bone-marrow hematopoietic cells stain strongly.	Those populations are listed as not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity may explain unexpected color (general IHC practice).	Compare a no-primary control and a reported positive population on the same run. Investigate the detection system before treating unexpected staining as TLN2 (general IHC practice).
The slide has broad, diffuse brown background.	Color outside interpretable cell boundaries can interfere with the cell-specific pattern reported by HPA (HPA tissue IHC; general IHC practice).	Inspect no-primary and detection controls, then review blocking, washes and detection development using the assay’s standard workflow (general IHC practice).
Kidney staining appears patchy across tubules.	HPA describes staining in a subset of renal tubules and specifically reports proximal-tubule microvilli at medium intensity (HPA tissue IHC).	Identify the tubule population and its staining compartment before judging the slide; do not require every tubule to stain (HPA tissue IHC).
ICC-IF shows focal-adhesion sites, but tissue IHC looks broadly cytoplasmic.	Both observations fit their respective reports: supported focal-adhesion and cytosol localization in ICC-IF, and cytoplasmic tissue staining (HPA subcellular; HPA tissue IHC).	Evaluate each assay against its own reported pattern and controls; do not require paraffin-section chromogenic IHC to resolve discrete focal-adhesion sites (general IHC practice).

Source: UniProt Q9Y4G6 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing TLN2 staining

IHC analysis of Talin 2 using anti-Talin 2 antibody (PB9961). Talin 2 was detected in a paraffin-embedded section of mouse intestine 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 1 μg/ml rabbit anti-Talin 2 Antibody (PB9961) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

PB9961 · IHCIHC analysis of Talin 2 using anti-Talin 2 antibody (PB9961). Talin 2 was detected in a paraffin-embedded section of mouse intestine tissue.

IHC analysis of Talin 2 using anti-Talin 2 antibody (PB9961). Talin 2 was detected in a paraffin-embedded section of rat intestine 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 1 μg/ml rabbit anti-Talin 2 Antibody (PB9961) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

PB9961 · IHCIHC analysis of Talin 2 using anti-Talin 2 antibody (PB9961). Talin 2 was detected in a paraffin-embedded section of rat intestine tissue. H

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000171914-TLN2/tissue/cerebral+cortex#imid_16496891

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000171914-TLN2/tissue/colon#imid_16496825

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000171914-TLN2/tissue/liver#imid_16496818

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000171914-TLN2/tissue/kidney#imid_16496811

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017194; green = TLN2, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for TLN2 IHC & IF

🧪Run cerebellum first: Purkinje cells should show cytoplasmic or membrane staining (HPA: Medium in Purkinje cells). Use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes); on the cerebellum slide, Purkinje cell nuclei and cell-free spaces should remain at background (UniProt Q9Y4G6: cytoplasmic and membrane localization; standard IHC control practice).

Positive control tissue: Caudate (Neuronal cells, HPA Medium)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TLN2 in A-431, U-251MG, U2OS, with annotated localisation: Focal adhesion sites (supported), Cytosol (supported) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality where applicable (caption: rabbit primary; standard IHC control practice); use TLN2 knockout material as a biological negative (standard IHC control practice). For chromogenic detection, quench endogenous peroxidase and assess endogenous biotin when using the caption’s avidin–biotin detection system (caption: SABC with DAB; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9961 paraffin-section caption does not state a fixative (caption: fixative not stated). The caption reports heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required for all specimens (caption: EDTA retrieval); neither frozen sections nor IF/ICC can be judged easier from the supplied evidence. For IF/ICC, HPA reports focal adhesion and cytosolic localization in imaged cell lines, so assess signal at those sites while accounting for specimen autofluorescence (HPA: subcellular localization; standard IF practice).

HPA tissue IHC evidence for TLN2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. 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
Caudate	Neuronal cells	Medium	Protein (IHC)	HPA →
Cerebellum	Purkinje cells - cytoplasm/membrane	Medium	Protein (IHC)	HPA →
Cerebral cortex	Neuronal cells	Medium	Protein (IHC)	HPA →
Heart muscle	Cardiomyocytes	Medium	Protein (IHC)	HPA →
Kidney	Proximal tubules (microvilli)	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Appendix	Glandular cells	Not detected	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced TLN2 IHC Tips

Troubleshoot TLN2 staining in paraffin sections by checking retrieval, compartment, cell type, and controls before comparing signal across samples.

Which retrieval conditions should I try first for TLN2 in paraffin sections?

Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet PB9961). The selected paraffin-section example used those conditions before overnight incubation at 4°C with 1 μg/ml primary antibody (datasheet PB9961). If staining is weak, compare a small range of heating times while keeping buffer, antibody concentration, detection, and tissue preparation constant; inspect morphology after each run (standard IHC practice). Include a known positive compartment in every run, such as renal proximal tubule microvilli (HPA tissue IHC), so stronger staining can be distinguished from broader, nonspecific color development (standard IHC practice).

Could fixation explain weak or patchy TLN2 staining?

Target-specific sensitivity of TLN2 to fixation is unknown from the supplied evidence (selected PB9961 caption: fixative not stated). Record the fixative, fixation duration, section thickness, and processing history for each block, then compare sections processed alike before changing antibody conditions (standard IHC practice). The selected PB9961 image establishes staining in a paraffin-embedded mouse intestine section with EDTA retrieval at pH 8.0, but does not identify its fixative (datasheet PB9961). If signal varies between blocks, repeat staining with a shared positive control and assess tissue preservation alongside staining; do not assign a TLN2-specific fixation effect without a controlled comparison (standard IHC practice).

Where should convincing TLN2 staining appear in tissue?

Evaluate TLN2 primarily in cytoplasmic and cell-associated patterns, including focal adhesion-associated regions; UniProt places it in cytoplasm, cytoskeleton, cell junctions, and synapses (UniProt Q9Y4G6 subcellular annotation). It has no transmembrane segment, so a membrane-edge signal need not represent a membrane-spanning protein (UniProt Q9Y4G6 topology). HPA reports cytoplasmic staining in neuronal cells and cardiomyocytes, plus proximal tubule microvilli staining in kidney (HPA tissue IHC). Compare the proposed compartment with neighboring cells and section morphology, and regard isolated nuclear color or uniform extracellular deposits as reasons to check background and controls (standard IHC practice).

How do I judge epitope-dependent TLN2 staining differences?

The record lists one TLN2 chain spanning residues 1–2542 and no annotated isoforms; it does not locate the PB9961 epitope (UniProt Q9Y4G6 record; supplied PB9961 caption). TLN2 contains a FERM domain at residues 88–406 and an I/LWEQ region at 2294–2533, so domain-level interpretations require a documented antibody epitope (UniProt Q9Y4G6 domains). Several phosphorylated residues are annotated, including serines 428 and 1023, but the supplied evidence does not establish whether modification changes this antibody's staining (UniProt Q9Y4G6 modified residues; supplied PB9961 caption). Compare retrieval and staining on matched sections before attributing a pattern difference to an isoform or modified epitope (standard IHC practice).

How should I assess TLN2 by IF alongside this IHC result?

Use a marker of the expected cell type in a separate fluorescence channel, such as a neuronal marker when examining neuronal staining (HPA tissue IHC), and include single-label controls when multiplexing (standard IF practice). Prefer a far-red fluorophore when the tissue shows strong shorter-wavelength autofluorescence, and assess unstained tissue before interpreting faint puncta (standard IF practice). TLN2 is cytosolic and associated with focal adhesion sites rather than spanning the membrane (HPA subcellular; UniProt Q9Y4G6 topology). Because the PB9961 epitope is unspecified, establish whether fixation and permeabilisation expose its cytoplasmic-facing epitope in IF; do not transfer the paraffin-section retrieval conditions directly to an IF specimen (supplied PB9961 caption; standard IF practice).

What should I change if TLN2 IHC has diffuse brown background?

The selected PB9961 paraffin-section example used 10% goat serum blocking, a biotinylated secondary antibody, streptavidin-biotin detection, and DAB (datasheet PB9961). Run a no-primary control and inspect whether color follows tissue edges, damaged regions, or the expected TLN2-positive cells (standard IHC practice; HPA tissue IHC). For peroxidase-based chromogenic IHC, check the peroxidase block and shorten DAB development if controls also darken; these are general workflow adjustments, not TLN2-specific requirements (standard IHC practice). If background persists, assess secondary-antibody reactivity and endogenous biotin with the chosen detection system before increasing the threshold used to call cells positive (standard IHC practice).

How can I compare TLN2 IHC across sections reliably? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell type and compartment before scoring, because reported TLN2 patterns include neuronal cytoplasm, cardiomyocytes, and renal proximal tubule microvilli (HPA tissue IHC). For comparable regions, record the percentage of positive cells and staining intensity, then calculate an H-score if intensity categories are reproducible; use positive-cell density per mm² when cell abundance itself is the outcome (standard IHC practice). Normalise to the number of eligible cells or the measured tissue area, and apply the same background threshold to all slides (standard IHC practice). Keep retrieval, DAB development, imaging, and region selection consistent, and report cell-type-specific results separately (standard IHC practice).

How do I distinguish true TLN2 signal from staining artefact?

A credible result combines the expected cell type with cytoplasmic or cell-associated staining: HPA reports neuronal cells, cardiomyocytes, and selected renal tubules, while UniProt places TLN2 at focal adhesions and in cytoplasm (HPA tissue IHC; UniProt Q9Y4G6 subcellular annotation). Treat isolated nuclear color, staining confined to section edges, and signal in necrotic areas as prompts to review morphology and controls (standard IHC practice). Compare the no-primary control to identify detection-system color, including possible endogenous enzyme signal during DAB development (standard IHC practice). HPA rates its tissue staining as Enhanced but notes only medium agreement with RNA data and pending external verification, so confirm consequential findings independently (HPA tissue IHC).

Boster reagents

Best TLN2 / Talin-2 IHC Antibodies

The IHC-validated TLN2 antibody has paraffin-section images from mouse and rat intestine and human intestinal cancer tissue (PB9961 image captions). No IF data are supplied (PB9961 catalog).

Real IHC data

Anti-Talin 2/TLN2 Antibody ®

Cat # PB9961

View product →

PB9961 is listed for IHC in human, mouse and rat samples (PB9961 catalog). Its IHC images show paraffin sections of mouse and rat intestine and human intestinal cancer tissue (PB9961 image captions).

Which to pick: Choose PB9961 for tissue IHC across human, mouse and rat samples (PB9961 catalog); its images support paraffin sections, and the fixative is unreported (PB9961 image captions). No SKU in this payload lists IF/ICC as an application, so there is no catalog-supported IF/ICC pick (PB9961 catalog).

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

References

UniProt Consortium. UniProt entry Q9Y4G6 (TLN2_HUMAN, Talin-2).
Human Protein Atlas. TLN2 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. TLN2 subcellular location (ICC-IF): Localized to the cytosol and focal adhesion sites..
Human Protein Atlas. TLN2 antibody validation summary (3 antibodies).
Patient genetics is linked to chronic wound microbiome composition and healing. PLoS pathogens 2020 — PMC7302439.
TLN2 functions as a tumor suppressor in clear cell renal cell carcinoma via inactivation of the Wnt/β-catenin signaling pathway. Translational andrology and urology 2022 — PMC8824820.
Male-specific epistasis between WWC1 and TLN2 genes is associated with Alzheimer's disease. Neurobiology of aging 2018 — PMC6769421.
PubMed PMID:12422219 — UniProt-cited evidence.
PubMed PMID:11527381 — UniProt-cited evidence.
PubMed PMID:16572171 — UniProt-cited evidence.