- HTA125 (See other available formats)
- HCDM listed
- Other Names
- Toll-like Receptor 4, TLR4, TLR-4, CD284
- Mouse IgG2a, κ
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- Product Citations
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Toll-like receptors are type I transmembrane signaling receptors. They are primordial pathogen-recognition proteins that function as sentinels for the innate immune system. TLR4, also known as CD284, is a 110 kD protein which is expressed on monocytes/macrophages, endothelial cells, and at low levels on B cells and granulocytes. In association with a secretory molecule, MD2, TLR4 has been recognized as critical for host recognition of bacterial LPS. HTA125 antibody is useful for flow cytometric analysis and is able to block LPS-induced cytokine production.Product Details
- Human, Cross-Reactivity: Guinea Pig
- Antibody Type
- Host Species
- Ba/F3 cell line expressing human TLR4
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 1 mM EDTA.
- The antibody was purified by chromatography and conjugated with TotalSeq™-A oligomer under optimal conditions. The solution is free of unconjugated DNA and unconjugated antibody.
- 0.5 mg/ml
- Storage & Handling
- The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.
PG - Quality tested
- Recommended Usage
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis and the oligomer sequence is confirmed by sequencing. For Proteogenomics TotalSeq™-A analysis, the suggested use of this reagent is ≤ 1.0 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
To maximize performance, centrifuge the antibody dilution (1.0 µg of antibody in 100 µl of staining buffer for every 1 million cells) before adding to the cells at 14,000xg at 2 - 8°C for 10 minutes. Carefully pipette out the liquid avoiding the bottom of the tube and add to the cell suspension.
Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.
- 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 production2,3,7,9. 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 LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm filtered) is recommended for functional assays (Cat. No. 312807). For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 312814) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin <0.01 EU/µg).
- Additional Product Notes
TotalSeq™ reagents are designed to profile protein expression at a single cell level following an optimized protocol similar to the CITE-seq workflow. A compatible single cell device (e.g. 10x Genomics Chromium System and Reagents) and sequencer (e.g. Illumina analyzers) are required. Please contact technical support for more information, or visit biolegend.com/totalseq.
The TotalSeq™-A barcode sequence associated with clone HTA125 is GCTTAGCTGTATCCG.
The flanking sequences are CCTTGGCACCCGAGAATTCCA, and the capture sequence, BAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA*A*A, B represents either C, G, or T, and * indicates a phosphorothioated bond, to prevent nuclease degradation.
The full oligomer sequence for this product, with the specific barcode in brackets is CCTTGGCACCCGAGAATTCCA [GCTTAGCTGTATCCG]BAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA*A*A.
(PubMed link indicates BioLegend citation)
- Skimazu R, et al. 1999. J. Exp. Med. 189:1777.
- Wang R, et al. 2003. Hybrid Hybridomics 22:357. (Block)
- Wang JE, et al. 2001. Infect. Immun. 69:2402. (Block)
- Ishihara S, et al. 2004 J. Immunol. 173:1406. (IHC)
- Kawahara T, et al. 2001 Infect. Immun. 69:4382.
- Jiang Q, et al. 2000. J. Immunol. 165:3541. (IF)
- Sugawara S, et al. 2001. Infect. Immun. 69:4951. (Block)
- Chavakis E, et al. 2007. Circ. Res. 100:204. PubMed
- Bhattacharyya S, et al. 2007. Am. J. Physiol. Gastrointest Liver Physiol. doi:10.1152/ajpgi.00149. (Block) PubMed
- Baumgarten G, et al. 2001. J. Infectious. Dis. 183:1617.
AB_2783183 (BioLegend Cat. No. 312817)
- Type I transmembrane protein, Toll-like receptor family/IL-1R superfamily, 110 kD
Monocytes/macrophages, endothelial cells, B cells and granulocytes
- 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
- View information about CD284 on UniProt.org
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