SEC23A / Protein transport protein Sec23A · IHC design guide

Design Immunohistochemistry for SEC23A

Plan SEC23A staining in paraffin sections with 2–5 μg/mL primary antibody (datasheet A05287-2). Expect cytoplasmic staining in several tissues, while interpreting specificity cautiously because HPA flags possible detection of proteins from multiple genes (HPA tissue IHC).

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

Immunohistochemistry protocol sheet for SEC23A (IHC for SEC23A): expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A05287-2, validated IHC image, and IHC protocol steps — Printable **SEC23A** IHC protocol sheet — expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A05287-2, controls and protocol steps. Open the full SEC23A IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

SEC23A 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 several tissues (HPA tissue IHC)
	Staining pattern	Cytoplasmic staining in neuronal, glandular and respiratory cells (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A05287-2)
	Positive control ⓘ	Cerebral cortex+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	Possible detection of proteins from multiple genes (HPA tissue IHC)
	Regulation	Ubiquitously expressed (UniProt)
	Isoform / epitope	2 isoforms; epitope differences unreported (UniProt)

Section 1

Recommended SEC23A IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol with three published SEC23A IHC methods (datasheet A05287-2; PMC5058693; PMC4974737; PMC10093042).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human placenta tissue; fixative not specified (datasheet A05287-2)
Fixation	Image fixative and duration unreported (datasheet A05287-2); 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 A05287-2); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A05287-2)
Primary antibody	Rabbit anti-SEC23A, 2-5μg/ml (datasheet A05287-2)
Primary incubation	Overnight at 4 °C (datasheet A05287-2)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05287-2)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	SEC23A-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A05287-2). A published alternative uses high-pressure citrate retrieval (PMC4974737 methods).

Section 2

What Is the Expected SEC23A Staining Pattern?

SEC23A is a cytosolic COPII component enriched at endoplasmic reticulum exit sites; it has no transmembrane segment (UniProt Q15436 localization and topology). In paraffin sections, expect cytoplasmic staining in cell populations such as cortical neurons, nasopharyngeal respiratory epithelium, and stomach glands (HPA tissue IHC: High). Treat this as a provisional pattern: HPA rates its tissue IHC evidence Uncertain because the antibody targets proteins from more than one gene (HPA tissue IHC).

What am I looking at on my slide?
Cytoplasmic staining in cortical neurons, respiratory epithelium, or stomach glands (HPA tissue IHC: High).	This matches the reported cell populations and cytoplasmic profile (HPA tissue IHC). A fine or partly punctate cytoplasmic pattern is compatible with ER exit sites and COPII vesicles (UniProt Q15436 localization); morphology alone cannot establish that every deposit is SEC23A.
Predominantly nuclear staining or isolated extracellular deposits.	These compartments do not match the reported cytosol, ER exit sites, and COPII vesicles (UniProt Q15436 localization; HPA subcellular). Review morphology and detection controls before scoring the signal as specific (general IHC practice).
Strong staining confined to adipocytes, cholangiocytes, or glomerular cells.	HPA reports these particular cell populations as Not detected (HPA tissue IHC). Investigate antibody cross-reactivity or endogenous detection activity (general IHC practice). An unexpected positive is suspicious, but HPA's Uncertain reliability prevents a definitive specificity call (HPA tissue IHC).
Diffuse color across tissue, stroma, and cells without a clear cytoplasmic pattern.	Uniform deposit obscures the expected cytoplasmic distribution (HPA tissue IHC; UniProt Q15436 localization). Compare no-primary and detection controls, then assess blocking, washing, and chromogen development (general IHC practice).
No signal in cortical neurons, nasopharyngeal respiratory epithelium, or stomach glands.	These are reported High-staining populations, so a blank reference section warrants a workflow check (HPA tissue IHC). Verify that tissue is present and intact, then review retrieval, primary antibody use, and detection controls (general IHC practice). HPA's Uncertain rating limits their value as definitive positives (HPA tissue IHC).

💡Expected SEC23A appearanceA plausible positive is cytoplasmic, potentially concentrated near ER exit sites, in HPA High-staining cell populations; dominant nuclear or extracellular color is suspect (UniProt Q15436 localization; HPA tissue IHC: High and Uncertain).

How each factor affects the staining
Cell population and tissue context	HPA reports High staining in cortical neurons, nasopharyngeal respiratory epithelium, and stomach glands, but Not detected in selected other cell populations (HPA tissue IHC). Compare like cell types when judging intensity; whole-organ averages can hide this variation.
Intracellular location and topology	SEC23A is associated with ER exit sites and COPII-coated vesicles and lacks a transmembrane segment (UniProt Q15436 localization and topology). Interpret a cytoplasmic pattern in that context; a surface-only pattern needs independent validation (general IHC practice).
IHC evidence quality	The reported tissue profile is Uncertain because the antibody targets proteins from more than one gene; HPA069974 is also rated Uncertain for IHC (HPA tissue IHC; HPA antibodies). Do not use staining intensity alone to claim SEC23A specificity.
Isoform interpretation	UniProt lists two SEC23A isoforms (UniProt Q15436 isoforms). The supplied sources give no epitope mapping or isoform-specific IHC result, so a chromogenic signal cannot identify which isoform contributes.
IF/ICC Q: Where should fluorescence appear?	A: HPA reports ER and cytosol as supported locations, with images from SiHa, U-251MG, and U2OS cells (HPA subcellular). Its subcellular summary cautions that the antibodies target proteins from multiple genes (HPA subcellular).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
High-staining reference cells are blank.	The expected cell population may be absent from the section, or an IHC step may have failed (HPA tissue IHC: High; general IHC practice).	Confirm cell identity and section integrity; check retrieval, primary antibody incubation, and detection controls (general IHC practice).
Most tissue is evenly brown.	Background from detection reagents or excessive chromogen development can obscure cellular localization (general IHC practice).	Review no-primary controls, blocking, washes, and development time; score only interpretable cellular staining (general IHC practice).
Signal is mainly nuclear.	Nuclear dominance conflicts with the reported cytosolic and ER-associated locations (UniProt Q15436 localization; HPA subcellular).	Check counterstain and tissue morphology, then compare detection controls and an independent specificity check (general IHC practice).
Adipocytes or cholangiocytes stain strongly.	HPA records these cell populations as Not detected, although its tissue IHC reliability is Uncertain (HPA tissue IHC).	Inspect no-primary controls for endogenous activity and seek independent confirmation before calling the signal SEC23A (general IHC practice).
Only some cells within a section stain.	HPA reports different staining levels by cell population; SEC23A is also described as ubiquitously expressed (HPA tissue IHC; UniProt Q15436 tissue specificity).	Identify and score each cell population separately; do not infer that unstained cells lack SEC23A protein (HPA tissue IHC: Uncertain).
A positive slide is used to claim a specific SEC23A isoform.	Two isoforms are listed, but the supplied sources provide no isoform-specific IHC evidence (UniProt Q15436 isoforms).	Report the result as provisional SEC23A-associated staining and use isoform-specific evidence for any isoform claim (UniProt Q15436 isoforms; HPA tissue IHC: Uncertain).

Source: UniProt Q15436 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing SEC23A staining

IHC analysis of SEC23A using anti-SEC23A antibody (A05287-2). SEC23A was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-SEC23A Antibody (A05287-2) 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.

A05287-2 · IHCIHC analysis of SEC23A using anti-SEC23A antibody (A05287-2). SEC23A was detected in paraffin-embedded section of human placenta tissue. Hea

IF analysis of SEC23A using anti-SEC23A antibody (A05287-2). SEC23A was detected in immunocytochemical section of HepG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-SEC23A Antibody (A05287-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.

A05287-2 · IFIF analysis of SEC23A using anti-SEC23A antibody (A05287-2). SEC23A was detected in immunocytochemical section of HepG2 cells. Enzyme antige

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100934-SEC23A/tissue/cerebral+cortex#imid_19720587

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100934-SEC23A/tissue/colon#imid_19720497

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100934-SEC23A/tissue/liver#imid_19720514

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000100934-SEC23A/tissue/kidney#imid_19720507

HPA · ICC-IF SiHaHuman Protein Atlas immunofluorescence, SiHa, antibody HPA069974; green = SEC23A, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for SEC23A IHC & IF

🧪Run cerebral cortex first and expect neuronal cells to stain (HPA: High in neuronal cells). Use smooth muscle cells as the negative tissue comparator (HPA: Not detected); on the cortex slide, treat cells without visible chromogen as internal background comparators, without assuming a particular non-neuronal cell type is SEC23A-negative.

Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SEC23A in SiHa, U-251MG, U2OS, with annotated localisation: Endoplasmic reticulum (supported), Cytosol (supported) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control; a species- and isotype-matched rabbit IgG control (caption: rabbit primary); and SEC23A-knockout material or a validated immunizing-peptide block as a biological specificity control (standard IHC practice). Quench endogenous peroxidase and check endogenous biotin for the biotin–SABC/DAB workflow (caption: biotinylated secondary and SABC/DAB); inspect no-primary cortex tissue for autofluorescence if using IF (standard IF practice).

⚠️Feasibility: A SEC23A-specific fixation window or fixation effect is unreported, and the selected A05287-2 paraffin-section caption does not state a fixative (caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but provides no comparison establishing that retrieval is required (caption: EDTA heat retrieval). Frozen-section ease and tissue IF ease are unreported; ICC-IF images exist for SiHa, U-251MG and U2OS (HPA subcellular), while cortical lipofuscin may complicate IF interpretation (standard IF practice).

HPA tissue IHC evidence for SEC23A

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Caution, targets protein from more than one gene. 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
Cerebral cortex	Neuronal cells	High	Protein (IHC)	HPA →
Nasopharynx	Respiratory epithelial cells	High	Protein (IHC)	HPA →
Stomach	Glandular cells	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →
Appendix	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Kidney	Cells in glomeruli	Not detected	Protein (IHC)	HPA →
Liver	Cholangiocytes	Not detected	Protein (IHC)	HPA →
Parathyroid gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Smooth muscle	Smooth muscle cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced SEC23A IHC Tips

Troubleshoot SEC23A staining in paraffin sections by checking retrieval, cytoplasmic localisation, controls and the limits of the available antibody evidence.

Which retrieval conditions should I try first for weak SEC23A staining in paraffin sections?

Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A05287-2). The selected paraffin section image used this condition before incubation with 2 μg/ml catalog antibody overnight at 4°C (datasheet A05287-2). If staining remains weak, compare a separately processed section using another routine retrieval buffer while holding antibody concentration, detection and development constant (standard IHC practice). Examine tissue integrity and background alongside signal, because excessive retrieval can damage morphology or increase nonspecific staining (standard IHC practice). Judge improvement in cytoplasmic staining consistent with SEC23A enrichment at ER exit sites, rather than by darker DAB alone (UniProt Q15436 localisation).

Could fixation explain weak or uneven SEC23A staining?

The selected tissue caption identifies a paraffin section but does not report its fixative, so target specific fixation sensitivity is unknown (datasheet A05287-2). Record the fixative and fixation duration for each specimen, then compare sections processed with the same EDTA pH 8.0 retrieval and detection conditions (datasheet A05287-2; standard IHC practice). Uneven fixation or processing can produce regional differences in staining, so assess preserved morphology before interpreting those differences as SEC23A biology (standard IHC practice). Keep section thickness and DAB development consistent across comparisons (standard IHC practice). Do not infer a preferred fixative from the reported HPA tissue staining pattern (HPA tissue IHC).

What SEC23A staining pattern is plausible in chromogenic IHC?

Expect predominantly cytoplasmic staining: SEC23A occurs in cytosol and on COPII coated vesicles and is enriched at ER exit sites (UniProt Q15436 localisation). A punctate or regionally concentrated cytoplasmic pattern can therefore be plausible, although DAB resolution may not identify individual exit sites (UniProt Q15436 localisation; standard IHC practice). SEC23A has no transmembrane segment, so a crisp cell surface outline alone is weak support for specific staining (UniProt Q15436 topology). Compare the suspect pattern with morphology and a secondary only control under the same development conditions (standard IHC practice). Treat isolated nuclear signal cautiously because the supplied localisation evidence supports ER and cytosol instead (HPA subcellular; UniProt Q15436 localisation).

How should I investigate epitope masking or isoform dependent staining?

SEC23A has 2 annotated isoforms, but the supplied evidence does not map this antibody’s epitope or establish isoform coverage (UniProt Q15436 isoforms; datasheet A05287-2). Check the antibody’s epitope documentation before interpreting a negative cell population as lacking both isoforms (standard IHC practice). Its annotated N terminal acetylation and Thr308 phosphorylation do not establish interference with this antibody without an epitope map and direct testing (UniProt Q15436 modified residues). Compare retrieval conditions on adjacent sections while keeping antibody concentration and DAB development fixed (standard IHC practice). If isoform discrimination matters, use an independently characterised reagent or orthogonal assay and report its demonstrated specificity (standard IHC practice).

How can I follow up an IHC pattern with multiplex IF?

For a separate IF experiment, pair SEC23A with a marker for the cell type identified by morphology in the IHC section, and include single stain controls (standard IF practice). Choose spectrally separated fluorophores after checking unstained tissue autofluorescence; a far red channel may improve contrast when shorter wavelengths show strong background (standard IF practice). Select permeabilisation that gives antibody access to the cytosolic side of the ER and COPII associated SEC23A, then verify that intracellular morphology remains intact (UniProt Q15436 topology and localisation; standard IF practice). HPA reports supported ER and cytosol localisation, but cautions that its antibodies target proteins from multiple genes (HPA subcellular). Do not transfer the paraffin section retrieval condition to IF without testing it (datasheet A05287-2; standard IF practice).

What should I check when DAB obscures cytoplasmic SEC23A signal?

Run a secondary only control and inspect whether diffuse colour persists without the primary antibody (standard IHC practice). The selected image used 10% goat serum blocking, a biotinylated secondary, an avidin biotin detection complex and DAB, so evaluate background at each applicable detection step (datasheet A05287-2). Include a peroxidase block as part of the general chromogenic workflow and check for endogenous biotin when using biotin based detection (standard IHC practice). Shorten DAB development or adjust primary antibody concentration systematically if tissue-wide colour masks cytoplasmic detail (standard IHC practice). Compare specific staining with ER and cytosol expectations rather than accepting uniform dark deposits (UniProt Q15436 localisation).

How should I quantify SEC23A staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define cell types and tissue regions before scoring, using the same section processing, imaging and DAB development conditions across samples (standard IHC practice). For cellular staining, report the percentage of positive cells and an intensity based H-score from 0–300; specify the intensity thresholds and whether scoring covers cytoplasm (standard IHC practice; UniProt Q15436 localisation). If counting positive cells by area, report density per mm² of viable tissue and normalise to viable tissue area or the number of evaluable cells (standard IHC practice). HPA describes low tissue specificity, so compare like cell populations across specimens (HPA tissue IHC). Exclude necrosis, folds and section edges before analysis (standard IHC practice).

How do I distinguish true SEC23A positivity from staining artefact?

Give greatest weight to reproducible cytoplasmic staining in morphologically intact cells, consistent with SEC23A at ER exit sites and in cytosol (UniProt Q15436 localisation; standard IHC practice). Isolated nuclear or sharply cell surface signal warrants scrutiny because neither is the supplied dominant localisation (UniProt Q15436 topology and localisation; HPA subcellular). Compare stained cells with the expected cell population in the section, and discount deposits concentrated at edges, folds or necrotic regions (standard IHC practice). Check secondary only and peroxidase blocked controls when DAB signal could reflect detection background or endogenous enzyme activity (standard IHC practice). HPA rates its tissue IHC evidence uncertain because the antibodies may recognise proteins from multiple genes; independent validation strengthens a SEC23A specific claim (HPA tissue IHC).

Boster reagents

Best SEC23A / Protein transport protein Sec23A IHC Antibodies

Anti-SEC23A antibody A05287-2 has IHC data from human paraffin-embedded placenta sections (IHC caption) and IF/ICC data from HepG2 cells (IF caption; catalog: ICC, IF).

Real IHC data

Anti-SEC23A Antibody ®

Cat # A05287-2

View product →

A05287-2 was used for chromogenic IHC on a human paraffin-embedded placenta section (IHC caption). The same SKU was used for fluorescence staining of HepG2 cells (IF caption; catalog: ICC, IF).

Which to pick: Choose A05287-2 for paraffin-section IHC: its own caption documents EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and DAB detection in human placenta; the fixative is unreported (IHC caption). Choose A05287-2 for IF/ICC based on its HepG2 fluorescence image and listed IF/ICC applications (IF caption; catalog: applications). For cross-species work, A05287-2 lists Human, Mouse and Rat reactivity, but the supplied IHC and IF images show human samples only; clonality is unreported (catalog: reactivity, clone; IHC and IF captions).

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

References

UniProt Consortium. UniProt entry Q15436 (SC23A_HUMAN, Protein transport protein Sec23A).
Human Protein Atlas. SEC23A tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. SEC23A subcellular location (ICC-IF): Localized to the endoplasmic reticulum and cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
Human Protein Atlas. SEC23A antibody validation summary (1 antibodies).
MicroRNA-375/SEC23A as biomarkers of the in vitro efficacy of vandetanib. Oncotarget 2016 — PMC5058693.
MicroRNA-21 promotes proliferation, migration, and invasion of colorectal cancer, and tumor growth associated with down-regulation of sec23a expression. BMC cancer 2016 — PMC4974737.
Increased SEC23A Expression Correlates with Poor Prognosis and Immune Infiltration in Stomach Adenocarcinoma. Cancers 2023 — PMC10093042.
SEC23A confers ER stress resistance in gastric cancer by forming the ER stress-SEC23A-autophagy negative feedback loop. Journal of experimental & clinical cancer research : CR 2023 — PMC10478313.
PubMed PMID:8898360 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:12508121 — UniProt-cited evidence.