Human Syndecan-4 Antibody

R&D Systems | Catalog # AF2918

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, Immunoprecipitation, Co-Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Syndecan-4 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Syndecan-4
Glu19-Glu145
Accession # P31431

Specificity

Detects human Syndecan-4 in direct ELISAs and Western blots. In direct ELISAs, approximately 25% cross-reactivity with recombinant mouse Syndecan-4 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Syndecan-4 Antibody

Syndecan-4 antibody in HeLa Human Cervical Epithelial Carcinoma Cell Line by Immunocytochemistry (ICC).

Syndecan‑4 in HeLa Human Cervical Epithelial Carcinoma Cell Line.

Syndecan-4 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human Syndecan-4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2918) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Syndecan-4 by Flow Cytometry

Detection of Syndecan-4 by Flow Cytometry

Relative expression of members of the Sdc4:RTK:ITG complex on nontumorigenic and neoplastic epithelial cells. Cell surface of expression of Sdc4 (mAb 8G3, red), Sdc2 (mAb 305515, orange), alpha 3 beta 1 (mAb P1B5, blue), alpha 6 beta 4 (mAb 3E1, green), EGFR (mAb EGFR.1, black), and RON (FAB6647F, purple) were analyzed by flow cytometry compared to nonspecific IgG (gray profile) on A, HNSCC (UM-SCC47, UM-SCC1) and TNBC (MDA-MB-231) cells or B, nontransformed HaCaT, HTE, or NOK epithelial cells. EGFR, epidermal growth factor receptor; HNSCC, head and neck squamous cell carcinoma; HTE, human tonsillar epithelial cell; NOK, normal oral keratinocyte; RON, recepteur d’origine nantais; Sdc2, syndecan-2; Sdc4, syndecan-4; TNBC, triple-negative breast cancer. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925822004690), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Syndecan-4 by Flow Cytometry

Detection of Syndecan-4 by Flow Cytometry

Relative expression of members of the Sdc4:RTK:ITG complex on nontumorigenic and neoplastic epithelial cells. Cell surface of expression of Sdc4 (mAb 8G3, red), Sdc2 (mAb 305515, orange), alpha 3 beta 1 (mAb P1B5, blue), alpha 6 beta 4 (mAb 3E1, green), EGFR (mAb EGFR.1, black), and RON (FAB6647F, purple) were analyzed by flow cytometry compared to nonspecific IgG (gray profile) on A, HNSCC (UM-SCC47, UM-SCC1) and TNBC (MDA-MB-231) cells or B, nontransformed HaCaT, HTE, or NOK epithelial cells. EGFR, epidermal growth factor receptor; HNSCC, head and neck squamous cell carcinoma; HTE, human tonsillar epithelial cell; NOK, normal oral keratinocyte; RON, recepteur d’origine nantais; Sdc2, syndecan-2; Sdc4, syndecan-4; TNBC, triple-negative breast cancer. Image collected and cropped by CiteAb from the following open publication (https://linkinghub.elsevier.com/retrieve/pii/S0021925822004690), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Syndecan-4 by Flow Cytometry

Detection of Syndecan-4 by Flow Cytometry

Effect of SDC4 knockdown (KD) or GAG inhibition on Delta spike internalization into Calu-3 cells. SDC4 KD in Calu-3 cells was performed using a lentiviral vector specific to human SDC4. (A) SDC4 expression levels were measured with imaging flow cytometry, as shown by the representative histograms and cellular images. Detected SDC4 levels of KD cells were normalized to WT Calu-3 cells as standards. The bars represent the mean + SEM of three independent experiments. Statistical significance vs. standards was assessed with ANOVA. * p < 0.05. (B,C) SDC4 KD and WT Calu-3 cells were treated with either the Delta spike protein (B) or the Delta PSV (C). For GAG inhibition, some of the WT Calu-3 cells were preincubated with heparin (200 ug/mL, 30 min before Delta treatment). Representative flow cytometry histograms and cellular images show the intracellular fluorescence of WT or SDC4 KD Calu-3 treated with Delta spike protein or PSV in the presence or absence of heparin. Scale bar = 20 μm. The effect of SDC4 KD or heparin treatment was expressed as percent inhibition, calculated with the following formula: ((X − Y)/X) × 100, where X is the fluorescence intensity obtained on controls treated with the Delta spike or Delta PSV without SDC4 KD or heparin, and Y is the fluorescence intensity obtained on cells receiving Delta spike or Delta PSV treatment, along with SDC4 KD or heparin. The bars represent the mean + SEM of three independent experiments. Statistical significance vs. controls was assessed with ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35054983), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Syndecan-4 by Flow Cytometry

Detection of Syndecan-4 by Flow Cytometry

Cellular uptake of the WT and Delta spike proteins into SDC4 deletion mutants. WT SDC4 transfectants and CBD, HSA and Si4 mutants (all tagged with GFP) were treated with the Delta spike. The cellular uptake was then analyzed with imaging flow cytometry by using fluorescently labeled specific antibodies against the His-tag of the recombinant spike proteins. (A) Representative flow cytometry histograms showing the intracellular fluorescence of WT SDC4 transfectants, CBD, HSA and Si4 mutants treated with the Delta spike protein (along with specific fluorescent antibodies). Scale bar = 20 μm. (B) Detected fluorescence intensities were normalized Delta spike-treated WT SDC4 transfectants as standards. The bars represent the mean ± SEM of four independent experiments (experimental data are represented as dots). Statistical significance vs. standards was assessed with ANOVA. *** p < 0.001. (C) Cellular images showing the intracellular fluorescence of WT SDC4 and CBD, HSA and Si4 mutants treated with the Delta spike protein. Scale bar = 20 μm. The indicated BDS values of SARS-CoV-2 and SDCs represent the mean + SEM of four independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35054983), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Syndecan-4 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HeLa human cervical epithelial carcinoma cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Western Blot

0.1 µg/mL
Sample: Recombinant Human Syndecan‑4 (Catalog # 2918-SD)

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Syndecan-4

Syndecan-4, previously known as amphiglycan or ryudocan, is a member of the syndecan family of Type 1 transmembrane proteins capable of carrying heparan sulfate (HS) and chondroitin sulfate glycosaminoglycans. The four vertebrate syndecans have two conserved cytoplasmic domains and divergent extracellular portions, except for HS attachment sites. Syndecan-4 is the most similar to Syndecan-2, but is more universally expressed and is found in virtually every cell type. Expression can be upregulated by TGF-beta 2 and in response to mechanical stress in smooth muscle, wound healing, arterial injury or acute myocardial infarction, probably in response to at least one inflammatory mediator (1, 2). Human Syndecan-4 is synthesized as a 198 amino acid (aa) core protein with an 18 aa signal sequence, a 127 aa extracellular domain containing three consensus Ser-Gly sequences for the attachment of HS side chains, a 25 aa transmembrane region and a 28 aa cytoplasmic tail (3). Human Syndecan-4 ECD shares approximately 79%, 78% and 81% aa identity with mouse, rat and porcine Syndecan-4 ECD, respectively. Addition of 20‑80 disaccharides per side chain adds considerably to the size of the 20 kDa core protein. Non-covalent homodimerization of Syndecan-4 is dependent on the transmembrane domain (4). The HS chains can bind fibronectin, SDF-1, antithrombin, FGF-2, midkine and tissue factor pathway inhibitor and can present FGF‑2 to its receptors (1, 2, 5). Proteolytic cleavage by plasmin, thrombin or a metalloproteinase may create a functional ectodomain (6‑8). Genetic disruption of the Syndecan-4 gene causes a mild phenotype, presumably due to compensation by other syndecans, but mice have an increase in placental thrombi as well as defects in wound healing and response to endotoxin shock (9, 10).

References

  1. Tkachenko, E. et al. (2005) Circ. Res. 96:488. 
  2. Oh, E.-S, and J. R. Couchman (2004) Mol. Cells 17:181.
  3. David, G. et al. (1992) J. Cell Biol. 118:961. 
  4. Choi, S. et al. (2005) J. Biol. Chem. 280:42573. 
  5. Charnaux, N. et al. (2005) FEBS J. 272:1937.  
  6. Schmidt, A. et al., J. Biol. Chem. 280:34441.
  7. Rauch, B. H. et al. (2005) J. Biol. Chem. 280:17507.
  8. Fitzgerald, M. L. et al. (2000) J. Cell Biol. 148:811.
  9. Ishiguro, K. et al. (2003) Glycoconj. J. 19:315.
  10. Echtermeyer, F. et al. (2001) J. Clin. Invest. 107:R9.

Alternate Names

Amphiglycan, Ryudocan, SDC4, Syndecan4

Entrez Gene IDs

6385 (Human); 20971 (Mouse)

Gene Symbol

SDC4

UniProt

P31431

Additional Syndecan-4 Products

Product Documents for Human Syndecan-4 Antibody

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Product Specific Notices for Human Syndecan-4 Antibody

For research use only

Citations for Human Syndecan-4 Antibody

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Protocols

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FAQs for Human Syndecan-4 Antibody

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  • Q: If Human Syndecan-4 is found on the membrane surface, and the immunogen is the extracellular portion of the protein, why does the ICC example for Catalog # AF2918 appear to have cytoplasmic staining?

    A: Our ICC protocol includes a permeabilization step prior to staining with the antibody, so the antibody will stain Syndecan-4 if present in the cytoplasm in addition to Syndecan-4 on the cell surface. 

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