Human CD14 Antibody

Catalog #: AF3831 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF3831-100
AF3831-SP

Detection of Human CD14 by Western Blot.

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Human CD14 Antibody Summary

Species Reactivity

Human

Specificity

Detects human CD14 in direct ELISAs.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Chinese Hamster Ovary cell line, CHO-derived human CD14 protein
Thr20-Cys352
Accession # P08571

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.5 µg/mL

Human PBMC

Simple Western

20 µg/mL

Human PBMC

Immunocytochemistry

5-25 µg/mL

Immersion fixed U937 human histiocytic lymphoma cells (positive) and absent in MOLT‑4 human acute lymphoblastic leukemia cells (negative)

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human CD14 by Western Blot. Western blot shows lysates of Human PBMC. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human CD14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3831) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for CD14 at approximately 53 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Simple Western

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Detection of Human CD14 by Simple Western^TM. Simple Western lane view shows lysates of Human PBMC, loaded at 0.2 mg/mL. A specific band was detected for CD14 at approximately 65 kDa (as indicated) using 20 µg/mL of Goat Anti-Human CD14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3831). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Immunocytochemistry

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Detection of CD14 in U937 cells (positive) and MOLT‑4 cells (negative). CD14 was detected in immersion fixed U937 human histiocytic lymphoma cells (positive) and absent in MOLT‑4 human acute lymphoblastic leukemia cells (negative) using Goat Anti-Human CD14 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3831) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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