Human CD14 Antibody
R&D Systems, part of Bio-Techne | Catalog # MAB3832
Conjugate
Catalog #
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
CyTOF-ready, Flow Cytometry, Immunohistochemistry, Neutralization, Western Blot
Cited:
ELISA Development, Flow Cytometry, Immunocytochemistry, Immunohistochemistry, Neutralization, Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # 134620
Product Specifications
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant human CD14
Thr20-Cys352
Accession # P08571
Thr20-Cys352
Accession # P08571
Specificity
Detects human CD14 in direct ELISAs and Western blots.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Human CD14 Antibody
Detection of Human CD14 by Western Blot.
Western blot shows lysates of human peripheral blood mononuclear cells (PBMC). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human CD14 Monoclonal Antibody (Catalog # MAB3832) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for CD14 at approximately 55 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Western Blot Buffer Group 1.
Detection of CD14 in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD14 Monoclonal Antibody (Catalog # MAB3832) followed by Goat anti-Mouse IgG APC-conjugated Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.
Detection of CD14 in human tonsil.
CD14 was detected in immersion fixed paraffin-embedded sections of human tonsil using Mouse Anti-Human CD14 Monoclonal Antibody (Catalog # MAB3832) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.Applications for Human CD14 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: Human peripheral blood monocytes
Sample: Human peripheral blood monocytes
Immunohistochemistry
5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil
Sample: Immersion fixed paraffin-embedded sections of human tonsil
Western Blot
2 µg/mL
Sample: Human peripheral blood mononuclear cells (PBMC)
Sample: Human peripheral blood mononuclear cells (PBMC)
Neutralization
Measured by its ability to neutralize lipopolysacharide (LPS)-induced TNF‑ alpha secretion in the THP-1 human acute monocytic leukemia cell line. At 10 µg/mL, this anti-hCD14 antibody will neutralize >60% of 0.5 ng/mL LPS-induced TNF-alpha secretion.
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 2 reviews rated 4.5 using MAB3832 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 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.
Background: CD14
CD14 is a 55 kDa cell surface glycoprotein that is preferentially expressed on monocytes/macrophages. The human CD14 cDNA encodes a 375 amino acid (aa) residue precursor protein with a 19 aa signal peptide and a C-terminal hydrophobic region characteristic for glycosylphosphatidyinositol (GPI)-anchored proteins. Human CD14 has four potential N-linked glycosylation sites and also bears O-linked carbohydrates. The amino acid sequence of human CD14 is approximately 65% identical with the mouse, rat, rabbit, and bovine proteins. CD14 is a pattern recognition receptor that binds lipopolysaccharides (LPS) and a variety of ligands derived from different microbial sources. The binding of CD14 with LPS is catalyzed by LPS-binding protein (LBP). The toll-like-receptors have also been implicated in the transduction of CD14-LPS signals. Similar to other GPI-anchored proteins, soluble CD14 can be released from the cell surface by phosphatidyinositol-specific phospholipase C. Soluble CD14 has been detected in serum and body fluids. High concentrations of soluble CD14 have been shown to inhibit LPS-mediated responses. However, soluble CD14 can also potentiate LPS response in cells that do not express cell surface CD14.
References
- Wright, S.D. et al. (1990) Science 249:1431.
- Pugin, J. et al. (1993) Proc. Natl. Acad. Sci. USA 90:2744.
- Beutler, B. (2000) Current Opinion in Immunology 12:20.
- Stelter, F. (2000) Chem. Immunol. 74:25.
Alternate Names
CD14
Gene Symbol
CD14
UniProt
Additional CD14 Products
Product Documents for Human CD14 Antibody
Product Specific Notices for Human CD14 Antibody
For research use only
Loading...
Loading...
Loading...
Loading...
Loading...