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48697
Toll-like Receptor Antibody Sampler Kit II
Primary Antibodies
Antibody Sampler Kit

Toll-like Receptor Antibody Sampler Kit II #48697

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

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.

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.

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.

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

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).

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.

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.

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

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

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

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.

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

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.

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).

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.

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.

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.

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 Includes Quantity Applications Reactivity MW(kDa) Isotype
Toll-like Receptor 1 Antibody 2209 20 µl
  • WB
H 86 Rabbit 
Toll-like Receptor 2 (D7G9Z) Rabbit mAb 12276 20 µl
  • WB
  • IP
H 90-105 Rabbit IgG
Toll-like Receptor 3 (D10F10) Rabbit mAb 6961 20 µl
  • WB
H 115-130 Rabbit IgG
Toll-like Receptor 6 (D1Z8B) Rabbit mAb 12717 20 µl
  • WB
  • IP
H M 90-110 Rabbit IgG
Toll-like Receptor 7 (D7) Rabbit mAb 5632 20 µl
  • WB
  • IP
H 140 Rabbit IgG
Toll-like Receptor 8 (D3Z6J) Rabbit mAb 11886 20 µl
  • WB
H 150 Rabbit IgG
Toll-like Receptor 9 (D9M9H) XP® Rabbit mAb 13674 20 µl
  • WB
  • IP
  • IF
  • F
H 130 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

For Research Use Only. Not For Use In Diagnostic Procedures.

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