Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of RAW 264.7 (Lane 1) and Hep G2 (Lane 2). The blot was probed with Anti-TLR4 Rabbit Polyclonal Antibody (Product# 48-2300, 2 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 95 kDa band corresponding to TLR4 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the C-terminal region of the mouse TLR4 protein (Accession# NP_067272, Q9QUK6), which is identical to rat and 94% homologous to human.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||20µg x 10^6 cells|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:50-1:200|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The protein encoded by this gene is a member of the Toll-like receptor (TLR) family which plays a fundamental role in pathogen recognition and activation of innate immunity. TLRs are highly conserved from Drosophila to humans and share structural and functional similarities. They recognize pathogen-associated molecular patterns (PAMPs) that are expressed on infectious agents, and mediate the production of cytokines necessary for the development of effective immunity. The various TLRs exhibit different patterns of expression. This receptor is most abundantly expressed in placenta, and in myelomonocytic subpopulation of the leukocytes. It has been implicated in signal transduction events induced by lipopolysaccharide (LPS) found in most gram-negative bacteria. Mutations in this gene have been associated with differences in LPS responsiveness. Also, several transcript variants of this gene have been found, but the protein coding potential of most of them is uncertain.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Vizantin inhibits endotoxin-mediated immune responses via the TLR 4/MD-2 complex.
48-2300 was used in immunocytochemistry and western blot to investigate the effect of vizantin on endotoxin-mediated immune responses
|Oda M,Yamamoto H,Shibutani M,Nakano M,Yabiku K,Tarui T,Kameyama N,Shirakawa D,Obayashi S,Watanabe N,Nakase H,Suenaga M,Matsunaga Y,Nagahama M,Takahashi H,Imagawa H,Kurosawa M,Terao Y,Nishizawa M,Sakurai J||Journal of immunology (Baltimore, Md. : 1950) (193:4507)||2014|
TLR4 activation promotes podocyte injury and interstitial fibrosis in diabetic nephropathy.
48-2300 was used in immunohistochemistry - paraffin section to examine the contribution of TLR4 to diabetic nephropathy
|Ma J,Chadban SJ,Zhao CY,Chen X,Kwan T,Panchapakesan U,Pollock CA,Wu H||PloS one (9:null)||2014|
|Mouse||1:50||Increased TLR4 expression in murine placentas after oral infection with periodontal pathogens.||Arce RM,Barros SP,Wacker B,Peters B,Moss K,Offenbacher S||Placenta (30:156)||2009|
|Human||Not Cited||Toll-like receptors TLR4, TLR5 and TLR9 on gastric carcinoma cells: an implication for interaction with Helicobacter pylori.||Schmausser B,Andrulis M,Endrich S,Müller-Hermelink HK,Eck M||International journal of medical microbiology : IJMM (295:179)||2005|
||Toll-like receptors TLR4, TLR5 and TLR9 on gastric carcinoma cells: an implication for interaction with Helicobacter pylori.||Schmausser B,Andrulis M,Endrich S,Müller-Hermelink HK,Eck M||International journal of medical microbiology : IJMM (295:179)||2005|