Order 3, Get Your 4th Free!* | Learn More >>
48697
Toll-like Receptor Antibody Sampler Kit II
Primary Antibodies
Antibody Sampler Kit

Toll-like Receptor Antibody Sampler Kit II #48697

Reviews ()
Citations (0)
Toll-like Receptor Antibody Sampler Kit II: Image 1

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with human TLR1, using Toll-like Receptor 1 Antibody.

Toll-like Receptor Antibody Sampler Kit II: Image 2

Immunoprecipitation of TLR2 from MUTZ-3 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Toll-like Receptor 2 (D7G9Z) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Toll-like Receptor 2 (D7G9Z) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 3

Western blot analysis of extracts from MCF 10A cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Toll-like Receptor 3 siRNA I #6236 (+), using Toll-like Receptor 3 (D10F10) Rabbit mAb #6961 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Toll-like Receptor 3 (D10F10) Rabbit mAb confirms silencing of Toll-like Receptor 3 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Toll-like Receptor Antibody Sampler Kit II: Image 4

Western blot analysis of extracts from THP-1 cells differentiated overnight with 80 nM TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 and Raw 264.7 cells using Toll-like Receptor 6 (D1Z8B) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 5

Western blot analysis of extracts from various cell lines using Toll-like Receptor 7 (D7) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 6

Western blot analysis of extracts from THP-1 cells differentiated with TPA #4174 (80 nM, 16 hr), untreated (-) or treated with Human Interferon-γ (hIFN-γ) #8901 (20 ng/mL, 8 hr; +), using Toll-like Receptor 8 (D3Z6J) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Toll-like Receptor Antibody Sampler Kit II: Image 7

Flow cytometric analysis of THP-1 cells (blue) and RPMI 8226 cells (green) using Toll-like Receptor 9 (D9M9H) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) was used as a secondary antibody.

Toll-like Receptor Antibody Sampler Kit II: Image 8

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Toll-like Receptor Antibody Sampler Kit II: Image 9

Western blot analysis of extracts from SR cells using Toll-like Receptor 1 Antibody.

Toll-like Receptor Antibody Sampler Kit II: Image 10

Western blot analysis of THP-1 and MUTZ-3 cells using Toll-like Receptor 2 (D7G9Z) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 11

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with human TLR3 (+), using Toll-like Receptor 3 (D10F10) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 12

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing HA-tagged full-length human TLR6 (hTLR6-HA; +) or transfected with a construct expressing HA-tagged full-length mouse TLR6 (mTLR6-HA; +), using Toll-like Receptor 6 (D1Z8B) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 13

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with human TLR7 (+), using Toll-like Receptor 7 (D7) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 14

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing HA-tagged full-length human TLR8 (hTLR8-HA; +), using Toll-like Receptor 8 (D3Z6J) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 15

Confocal immunofluorescent analysis of RPMI 8226 (left) and THP-1 (right) cells using Toll-like Receptor 9 (D9M9H) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Toll-like Receptor Antibody Sampler Kit II: Image 16

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing HA-tagged full-length human TLR2 (hTLR2-HA; +), using Toll-like Receptor 2 (D7G9Z) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 17

Western blot analysis of extracts from HT-29 cells, untreated or following transfection with pIpC (100 μg/ml; overnight), using Toll-like Receptor 3 (D10F10) Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 18

Immunoprecipitation of Toll-like receptor 9 from Ramos cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Toll-like Receptor 9 (D9M9H) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Toll-like Receptor 9 (D9M9H) XP® Rabbit mAb.

Toll-like Receptor Antibody Sampler Kit II: Image 19

Western blot analysis of extracts from various cell lines using Toll-like Receptor 9 (D9M9H) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

To Purchase # 48697T
Product # Size Price
48697T
1 Kit  (6 x 20 µl) $ 445

Product Description

The Toll-like Receptor Antibody Sampler Kit II provides an economical means of detecting expression of various Toll-like receptors (TLRs). The kit contains enough primary and secondary antibodies to perform at least two western blot experiments.

Specificity / Sensitivity

Each antibody in the Toll-like Receptor Antibody Sampler Kit II detects endogenous levels of its target protein. Toll-like Receptor 3 (D10F10) Rabbit mAb detects a 75 kDa protein of unknown origin in some cell lines and tissues. Toll-like Receptor 6 (D1Z8B) Rabbit mAb detects a 72 kDa protein of unknown origin. Toll-like Receptor 6 (D1Z8B) Rabbit mAb is not approved for IP in mouse samples. Toll-like Receptor 8 (D3Z6J) Rabbit mAb detects 30 kDa and 37 kDa proteins of unknown origin. Toll-like Receptor 9 (D9M9H) XP® Rabbit mAb is predicted to react with known full-length isoforms of TLR9, but not with the cleaved TLR9 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to residues surrounding Val303 of human TLR2 protein, Val495 of human TLR3 protein, Pro47 of human TLR6 protein, Pro141 of human TLR8 protein, Pro450 of human TLR9 protein, or recombinant protein specific to the leucine-rich repeats within human TLR7 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues at the amino terminus of human TLR1 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Members of the Toll-like receptor (TLR) family, named for the closely related Toll receptor in Drosophila, play a pivotal role in innate immune responses (1-4). TLRs recognize conserved motifs found in various pathogens and mediate defense responses (5-7). TLR1, TLR2, TLR4, TLR5, TLR6, and TLR11 are localized to the plasma membrane, while TLR3, TLR7, TLR8, and TLR9 are localized to intracellular membranes including endosomal membranes. Triggering of the TLR pathway leads to the activation of NF-κB and subsequent regulation of immune and inflammatory genes (4). The TLRs and members of the IL-1 receptor family share a conserved stretch of approximately 200 amino acids known as the Toll/Interleukin-1 receptor (TIR) domain (1). Upon activation, TLRs associate with a number of cytoplasmic adaptor proteins containing TIR domains, including myeloid differentiation factor 88 (MyD88), MyD88-adaptor-like/TIR-associated protein (MAL/TIRAP), Toll-receptor-associated activator of interferon (TRIF), and Toll-receptor-associated molecule (TRAM) (8-10). This association leads to the recruitment and activation of IRAK1 and IRAK4, which form a complex with TRAF6 to activate TAK1 and IKK (8,11-14). Activation of IKK leads to the degradation of IκB, which normally maintains NF-κB in an inactive state by sequestering it in the cytoplasm. TLR1 and TLR6 associate with TLR2 to cooperatively mediate response to bacterial lipoproteins and fungal zymosan (6,15). TLR3 is an endosomal TLR that recognizes double-stranded RNA derived from viruses (7). TLR7 and TLR8 recognize single-stranded viral RNA and are also activated by synthetic imidazoquinoline compounds including R-848 (16,17). TLR9 recognizes unmethylated CpG motifs present on bacterial DNA (18).

  1. Akira, S. (2003) J Biol Chem 278, 38105-8.
  2. Beutler, B. (2004) Nature 430, 257-63.
  3. Dunne, A. and O'Neill, L.A. (2003) Sci STKE 2003, re3.
  4. Medzhitov, R. et al. (1997) Nature 388, 394-7.
  5. Schwandner, R. et al. (1999) J Biol Chem 274, 17406-9.
  6. Takeuchi, O. et al. (1999) Immunity 11, 443-51.
  7. Alexopoulou, L. et al. (2001) Nature 413, 732-8.
  8. Zhang, F.X. et al. (1999) J Biol Chem 274, 7611-4.
  9. Horng, T. et al. (2001) Nat Immunol 2, 835-41.
  10. Oshiumi, H. et al. (2003) Nat Immunol 4, 161-7.
  11. Muzio, M. et al. (1997) Science 278, 1612-5.
  12. Wesche, H. et al. (1997) Immunity 7, 837-47.
  13. Suzuki, N. et al. (2002) Nature 416, 750-6.
  14. Irie, T. et al. (2000) FEBS Lett 467, 160-4.
  15. Ozinsky, A. et al. (2000) Proc Natl Acad Sci U S A 97, 13766-71.
  16. Heil, F. et al. (2004) Science 303, 1526-9.
  17. Jurk, M. et al. (2002) Nat Immunol 3, 499.
  18. Hemmi, H. et al. (2000) Nature 408, 740-5.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.