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Purified Rat Anti-Mouse CD284/MD-2 Complex
Purified Rat Anti-Mouse CD284/MD-2 Complex
Expression of TLR4-MD-2 on peritoneal macrophages. Thioglycollate-elicited peritoneal macrophages from BALB/c mice were stained with either purified rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553927, left panel) or purified mAb MTS510 (right panel), in the presence of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142). Staining was detected by biotinylated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894) then Streptavidin-PE (Cat. No. 554061). The macrophages were identified by staining with FITC-conjugated anti-mouse CD11b (Integrin αM chain) mAb M1/70 (Cat. No. 557396/553310). The total viable leukocytes are displayed. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Expression of TLR4-MD-2 on peritoneal macrophages. Thioglycollate-elicited peritoneal macrophages from BALB/c mice were stained with either purified rat IgG2a, κ isotype control mAb R35-95 (Cat. No. 553927, left panel) or purified mAb MTS510 (right panel), in the presence of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142). Staining was detected by biotinylated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894) then Streptavidin-PE (Cat. No. 554061). The macrophages were identified by staining with FITC-conjugated anti-mouse CD11b (Integrin αM chain) mAb M1/70 (Cat. No. 557396/553310). The total viable leukocytes are displayed. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
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BD Pharmingen™
TLR4/MD-2 Complex
Mouse (QC Testing)
Rat IgG2a, κ
Mouse Pro-B cell line Ba/F3 expressing transfected BALB/c mouse TLR4 and MD-2
Flow cytometry (Routinely Tested), Blocking, Immunoprecipitation (Reported)
0.5 mg/ml
21898
AB_397072
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


推荐的实验流程

Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) may help to reduce non-specific binding to cells bearing Fcγ receptors. If Mouse BD Fc Block™ is used, then it is important that the second-step anti-rat IgG antibody does not cross-react with the 2.4G2 mAb (Rat IgG2b, κ); we recommend the use of biotinylated anti-rat IgG2a mAb RG7/1.30 (Cat. No. 553894) with a "bright" third-step reagent, such as Streptavidin-PE (Cat. No. 554061). Other reported applications include immunoprecipitation1, and blocking of LPS-induced NF-κB activation or TNF production.

商品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
558293 Rev. 1
抗体详情
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MTS510

The MTS510 antibody reacts with the molecular complex of Toll-Like Receptor 4 and MD-2 (TLR4-MD-2) which is expressed on LPS responsive macrophages. TLR4, a member of the Toll-Like Receptor Family, has been renamed as CD284 and identified to be the transmembrane signal-transducing portion of the receptor for LPS. TLR4 associates on the cell surface with CD14 and MD-2, a 0.7 kDa molecule which is anchored to the membrane via its physical association with TLR4. The association of MD-2 with TLR4 is required for recognition of LPS and the anti-mitotic compound Taxol, which mimics the action of LPS on mouse cells. MTS510 mAb detects TLR4-MD-2 on the surface of thioglycollate-elicited macrophages from all mouse strains tested (ie, BALB/c, C57BL/6, C3H/HeJ, C3H/HeN, and DBA/1), including the C3H/HeJ strain which expresses an LPS-resistant mutant TLR4. Expression of TLR4-MD-2 is down-regulated on peritoneal macrophages after exposure to LPS, correlating with the occurrence of LPS tolerance. TLR4-MD-2 is not detected on splenocytes or thymocytes.

558293 Rev. 1
格式详情
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
558293 Rev.1
报价单和参考
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研发参考 (6)

  1. Akashi S, Saitoh S, Wakabayashi Y, et al. Lipopolysaccharide interaction with cell surface Toll-like receptor 4-MD-2: higher affinity than that with MD-2 or CD14. J Exp Med. 2003; 198(7):1035-1042. (Clone-specific: Immunoprecipitation). 查看参考
  2. Akashi S, Shimazu R, Ogata H, et al. Cutting edge: cell surface expression and lipopolysaccharide signaling via the toll-like receptor 4-MD-2 complex on mouse peritoneal macrophages. J Immunol. 2000; 164(7):3471-3475. (Immunogen: Blocking, Immunoprecipitation). 查看参考
  3. Beutler B, Poltorak A. The sole gateway to endotoxin response: how LPS was identified as Tlr4, and its role in innate immunity. Drug Metab Dispos. 2001; 29(4):474-478. (Biology). 查看参考
  4. Kawasaki K, Nogawa H, Nishijima M. Identification of mouse MD-2 residues important for forming the cell surface TLR4-MD-2 complex recognized by anti-TLR4-MD-2 antibodies, and for conferring LPS and taxol responsiveness on mouse TLR4 by alanine-scanning mutagenesis. J Immunol. 2003; 170(1):413-420. (Biology). 查看参考
  5. Nomura F, Akashi S, Sakao Y, et al. Cutting edge: endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface toll-like receptor 4 expression. J Immunol. 2000; 164(7):3476-3479. (Immunogen). 查看参考
  6. Shimazu R, Akashi S, Ogata H, et al. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med. 1999; 189(11):1777-1782. (Biology). 查看参考
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558293 Rev. 1

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