Human CD14 Biotinylated Antibody

R&D Systems | Catalog # BAF383

R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Detection (Matched Antibody Pair)

Cited:

Immunohistochemistry, Western Blot, Affinity Purification, Cell Selection, ELISA Detection, ELISA Development, ELISA Microarray Development

Label

Biotin

Antibody Source

Polyclonal Sheep IgG
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 ELISAs and Western blots. In sandwich ELISAs, less than 0.2% cross-reactivity with recombinant mouse CD14 and Lipopolysaccharide is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human CD14 Biotinylated Antibody

CD14 antibody in Human Lymph Node by Immunohistochemistry (IHC-P).

CD14 in Human Lymph Node.

CD14 was detected in immersion fixed paraffin-embedded sections of human lymph node using 10 µg/mL Sheep Anti-Human CD14 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF383) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-AEC Cell & Tissue Staining Kit (red; CTS020) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of CD14 in THP‑1 (Positive) and MOLT‑4 (Negative).

CD14 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line (Positive) and absent in MOLT‑4 human acute lymphoblastic leukemia cell line (Negative) using Sheep Anti-Human CD14 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF383) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of CD14 in U937(Positive) and MOLT‑4 (Negative).

CD14 was detected in immersion fixed U937 human histiocytic lymphoma cell line (Positive) and absent in MOLT‑4 human acute lymphoblastic leukemia cell line (Negative) using Sheep Anti-Human CD14 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF383) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Human CD14 Biotinylated Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line (Positive), MOLT‑4 human acute lymphoblastic leukemia cell line (Negative) and U937 human histiocytic lymphoma cell line (Positive).

Western Blot

0.1 µg/mL
Sample: Recombinant Human CD14 (Catalog # 383-CD)

Human CD14 Sandwich Immunoassay

ELISA Detection (Matched Antibody Pair)
Recommended Concentration: 0.1-0.4 µg/mL
Use in combination with these reagents:
  • Capture Reagent: Human CD14 Antibody (Catalog # MAB3833)
  • Capture Reagent: Human CD14 Antibody (Catalog # MAB3833R)
  • Standard: Recombinant Human CD14 Protein (Catalog # 383-CD)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using BAF383 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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 Biotinylated Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CD14 Biotinylated Antibody

For research use only

Citations for Human CD14 Biotinylated Antibody

Customer Reviews for Human CD14 Biotinylated Antibody (2)

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  • Human CD14 Biotinylated Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Whole blood
    Species: Human
    Verified Customer | Posted 10/14/2021
    The 1:1000 dilution secondary anti-human CD14 Biotinylated Antibody was added.
    Human CD14 Biotinylated Antibody BAF383
  • Human CD14 Biotinylated Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Peripheral blood cells
    Species: Human
    Verified Customer | Posted 07/27/2016

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Protocols

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