Anti-TLR4 antibody (bs-20595R)

Anti-TLR4 antibody

DATASHEET

Host:Rabbit

Target Protein:TLR4

IR:Immunogen Range:121-200/839

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:7099

Swiss Prot:O00206

Source:KLH conjugated synthetic peptide derived from human TLR4:121-200/839 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:The Toll-like receptor 4 by activating natural immunity, specific immune response involved in the start-up, Toll-like receptor 4 as an important signal transduction transmembrane receptor involved in the toxin-induced inflammation in the pathological process, its mechanisms of control On a growing concern. all regions were either double-stranded or sequenced with an alternatechemistry or covered by high quality data(i.e., phred quality >=30); an attempt was made to resolve all sequencing problems, such as compressions and repeats; all regions were covered by at least one subclone; and the assembly was confirmed by restriction digest, except on the rare occasion of the clone being a YAC. The following abbreviations are used to associate primary accession numbers given in the feature table with their source databases: Em:, EMBL; Sw:, SWISSPROT; Tr:, TREMBL; Wp:, WORMPEP; Information on the WORMPEP database can be found at. TLR-4 plays an important role in microvascular leakage and leukocyte adhesion under the inflammatory condition associated with nonseptic thermal injury.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
Flow-Cyt(1μg/Test)
ICC(1:100)

Cross Reactive Species:Human
Mouse
Rat
Pig
Cow
Sheep
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample:
Lane 1: H9C2 (Human) Cell Lysate at 30 ug
Lane 2: K562 (Human) Cell Lysate at 30 ug
Primary: Anti-TLR4 (bs-20595R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 110 kD
Observed band size: 110 kD

Tissue/cell:U2OS cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (TLR4) polyclonal Antibody, Unconjugated (bs-20595R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-TLR4 antibody (bs-20595R-PE-Cy7)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.

Blank control:U2OS.
Primary Antibody (green line): Rabbit Anti-TLR4 antibody (bs-20595R-PE-Cy7)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Blank control:RAW264.7.
Primary Antibody (green line): Rabbit Anti-TLR4 antibody (bs-20595R-PE-Cy7)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.

Blank control:RAW264.7.
Primary Antibody (green line): Rabbit Anti-TLR4 antibody (bs-20595R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.

Blank control:Jurkat.
Primary Antibody (green line): Rabbit Anti-TLR4 antibody (bs-20595R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.

Blank control:HUVEC.
Primary Antibody (green line): Rabbit Anti-TLR4 antibody (bs-20595R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1%PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.

Blank control:U2OS.
Primary Antibody (green line): Rabbit Anti-TLR4 antibody (bs-20595R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.