Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization, Flow Cytometry, CyTOF-ready

Cited:

Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, ELISA Development

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 134620
View all CD14 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human CD14
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 antibody in Human PBMCs antibody by Flow Cytometry.

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

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Western Blot

2 µg/mL
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.

Reviewed Applications

Read 2 reviews rated 4.5 using MAB3832 in the following applications:

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Flow Cytometry

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.

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: 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

  1. Wright, S.D. et al. (1990) Science 249:1431.
  2. Pugin, J. et al. (1993) Proc. Natl. Acad. Sci. USA 90:2744.
  3. Beutler, B. (2000) Current Opinion in Immunology 12:20.
  4. Stelter, F. (2000) Chem. Immunol. 74:25.

Alternate Names

CD14

Entrez Gene IDs

929 (Human); 12475 (Mouse); 60350 (Rat); 100037938 (Porcine)

Gene Symbol

CD14

UniProt

P08571

Additional CD14 Products

Product Documents for Human CD14 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CD14 Antibody

For research use only

Citations for Human CD14 Antibody

Customer Reviews for Human CD14 Antibody (2)

4.5 out of 5
2 Customer Ratings
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Showing  1 - 2 of 2 reviews Showing All
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  • Human CD14 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: THP-1 human acute monocytic leukemia cell line
    Species: Human
    Verified Customer | Posted 10/12/2021
    Cells lysed in NP-40 lysis buffer. TBST-150mM NaCl. Blocked with 5% milk TBST. Antibody diluted in 2% milk TBST 1:100 (5ug/ml). Product works better under non-reducing conditions.
    Human CD14 Antibody MAB3832
  • Human CD14 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: wheat extracts
    Species: wheat
    Verified Customer | Posted 03/29/2021
    we use this antibody to see interactions with CD14 recombinant protein, it showed a very good affinity towards CD14 protein in direct ELISA.
    Human CD14 Antibody MAB3832

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Protocols

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