Ultra-LEAF™ Purified anti-human CD284 (TLR4) Antibody

Product Details

Verified Reactivity

Human

Reported Reactivity

Guinea Pig

Antibody Type

Monoclonal

Host Species

Mouse

Immunogen

Ba/F3 cell line expressing human TLR4

Formulation

0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.

Endotoxin Level

Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.

Preparation

The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.

Concentration

The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.

Application

FC - Quality tested
IHC-F, ICC, WB, Block - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 2.0 µg per million cells in 100 µl volume or 100 µl of whole blood.  It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections⁴, immunofluorescence microscopy⁶, Western blotting¹⁰, and in vitro blocking of LPS-induced cytokine production^2,3,7,9. This clone was tested in-house and does not work on formalin fixed paraffin-embedded (FFPE) tissue. For most successful immunofluorescent staining results, it may be important to maximize signal over background by using a relatively bright fluorochrome-antibody conjugate (Cat. No. 312806) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated antibody (Cat. No. 312804) or biotinylated anti-mouse IgG second step (Cat. No. 405303), followed by SAv-PE (Cat. No. 405204). The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 312813 & 312814). 

Application References

(PubMed link indicates BioLegend citation)

1. Skimazu R, et al. 1999. J. Exp. Med. 189:1777.
2. Wang R, et al. 2003. Hybrid Hybridomics 22:357. (Block)
3. Wang JE, et al. 2001. Infect. Immun. 69:2402. (Block)
4. Ishihara S, et al. 2004 J. Immunol. 173:1406. (IHC)
5. Kawahara T, et al. 2001 Infect. Immun. 69:4382.
6. Jiang Q, et al. 2000. J. Immunol. 165:3541. (IF)
7. Sugawara S, et al. 2001. Infect. Immun. 69:4951. (Block)
8. Chavakis E, et al. 2007. Circ. Res. 100:204. PubMed
9. Bhattacharyya S, et al. 2007. Am. J. Physiol. Gastrointest Liver Physiol. doi:10.1152/ajpgi.00149. (Block) PubMed
10. Baumgarten G, et al. 2001. J. Infectious. Dis. 183:1617.

Product Citations

1. Murata K, et al. 2023. J Immunol. 310:210. PubMed
2. Lee Y, et al. 2022. Front Immunol. 12:780210. PubMed

RRID

AB_2561485 (BioLegend Cat. No. 312813)
AB_2561485 (BioLegend Cat. No. 312814)

Antigen Details

Structure

Type I transmembrane protein, Toll-like receptor family/IL-1R superfamily, 110 kD

Distribution

Monocytes/macrophages, endothelial cells, B cells and granulocytes

Function

Activates the innate immune system against Gram-negative bacterial pathogens by recognition of LPS

Ligand/Receptor

Bacterial LPS

Cell Type

B cells, Endothelial cells, Granulocytes, Macrophages, Monocytes, Tregs

Biology Area

Cell Biology, Immunology, Innate Immunity, Neuroinflammation, Neuroscience, Neuroscience Cell Markers

Molecular Family

CD Molecules, Toll Like Receptors

Antigen References

1. Skimazu R, et al. 1999. J. Exp. Med. 189:1777.

Gene ID

7099 View all products for this Gene ID

UniProt

View information about CD284 on UniProt.org

Documentation

• Technical data sheet
• Certificate of Analysis
• Material Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

Do you guarantee that your antibodies are totally pathogen free?

BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.

Does BioLegend test each Ultra-LEAF™ antibody by functional assay?

No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.

Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?

No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.

Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?

We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.

Other Formats

Certificate of Analysis