TLR4 Antibody (76B357.1)

Images

 
Orthogonal Strategies: Dual RNAscope ISH-IHC: TLR4 Antibody (76B357.1) [NB100-56566] - Formalin-fixed paraffin-embedded tissue sections of human tonsil were probed for TLR4 mRNA (ACD RNAScope Probe, catalog ...read more
Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. Double-stained immunofluorescence for TLR4 (green) and ...read more
Western Blot: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using 2 ug/ml on (A) human intestine and 6 ug/ml on (B) mouse intestine and C) rat intestine lysate.
Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using the Alexa Fluor (R) 647 conjugate of NBP2-27149. TLR4 expression on monocytes from human peripheral blood: PBMC were stained, in a 2 color flow ...read more
Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. IHC staining of colon adenocarcinoma for TLR4, vimentin, and ...read more
Flow (Intracellular): TLR4 Antibody (76B357.1) [NB100-56566] - Analysis using PE conjugate of NBP2-27149. An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149PE (blue) and an ...read more
Immunohistochemistry: TLR4 Antibody (76B357.1) [NB100-56566] - Immunofluorescent staining of TMAs. Representative tissue cores from normal (I), adenomatous polyps (II), and CRC (III and IV) are shown. Image collected ...read more
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of Rat's salivary gland tissue section at 1:100 dilution. The antibody generated a membrane-cytoplasmic staining in the tissue with ...read more
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Tissue section of normal human skin at 5 ug/ml. Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells ...read more
Flow Cytometry: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of formaldehyde fixed THP-1 cells (human monocytic leukemia cells) using 2 ug/10^6 cells TLR4 antibody (clone 76B357.1) with detection employing a donkey ...read more
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human skin stained with at 5 ug/ml, peroxidase-conjugate and DAB chromogen. Staining of formalin-fixed tissues is enhanced by boiling tissue ...read more
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Analysis of TLR4 in paraffin-embedded formalin-fixed human colon tissue using an isotype control (top) and NB100-56566 (bottom) at 5 ug/ml.
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) [NB100-56566] - Human testis tissue stained using 5 ug/ml.
Immunohistochemistry-Frozen: TLR4 Antibody (76B357.1) [NB100-56566] - This image is TLR4(green) and nucleus(blue) at area postrema of the adult male mouse brain, x20 magnification. Primary antibody diluted 1:500. This ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHaSpecies Glossary
Applications WB, ChIP, DB, ELISA, Flow, Flow-CS, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, B/N, CyTOF-ready, Dual ISH-IHC, ELISA(Cap), KD, KO
Clone
76B357.1
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1.0 mg/ml
Validated by:
       

Orthogonal Strategies

 

Order Details

TLR4 Antibody (76B357.1) Summary

Immunogen
This antibody was developed against a portion of amino acids 100-200 of human TLR4 (NP_612564).
Predicted Species
Chinese Hamster (100%), Baboon (100%). Backed by our 100% Guarantee.
Isotype
IgG2b Kappa
Clonality
Monoclonal
Host
Mouse
Gene
TLR4
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1-3 ug/ml
  • Chromatin Immunoprecipitation 1:10-1:500
  • Dot Blot
  • ELISA
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:500
  • Immunohistochemistry-Paraffin 5 ug/ml
  • In vitro assay
  • Block/Neutralize
  • CyTOF-ready
  • Dual RNAscope ISH-IHC
  • ELISA Capture (Matched Antibody Pair) 0.5 ug
  • Flow (Cell Surface)
  • Flow (Intracellular)
  • Knockdown Validated
  • Knockout Validated
Application Notes
Flow (Intracellular) use reported in literature (See Cohen et al, 2008); Immunofluorescence (See Nowicki et al, 2009); IHC-F (Nowicki et al, 2012); Flow (cell surface) use reported in scientific literature (PMID: 23796194). Use in chromatin immunoprecipitation reported in scientific literature (PMID: 21966468). Use In vitro reported in scientific literature (PMID: 26446256). Use in blocking/neutralizing reported in scientific literature (PMID 26063617). Use in ELISA reported in scientific literature (PMID: 30139760). Use in Dot blot reported in scientific literature (PMID: 27248820). Use in Knockout Validated reported in scientific literature (PMID: 31125703). Knockdown validation (PMID: 31766635). This TLR4 antibody is CyTOF ready.
Agonist
Antagonist
Reviewed Applications
Read 5 Reviews rated 4.2
using
NB100-56566 in the following applications:

Publications
Read Publications using
NB100-56566 in the following applications:

Reactivity Notes

Ground squirrel reactivity reported by a customer review. Mammal reactivity reported in scientific literature (PMID: 25130694). Porcine reactivity reported in scientific literature (PMID: 29941006). Bovine reactivity reported in scientific literature (PMID: 19528595). Sheep (85%), Cat, Equine (78%). Mammal reactivity reported in scientific literature (PMID: 25130694).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for TLR4 Antibody (76B357.1)

  • 76B357.1
  • ARMD10
  • CD284 antigen
  • CD284
  • hTollhomolog of Drosophila toll
  • tlr4 76B357.1
  • TLR4 facs
  • TLR4 flow cytometry
  • TLR4 human
  • TLR4 ihc
  • TLR4 western blot
  • TLR4
  • TOLL
  • toll-like receptor 4

Background

The Toll-like receptor (TLR) family in mammal comprises a family of transmembrane proteins characterized by multiple copies of leucine rich repeats in the extracellular domain and IL-1 receptor motif in the cytoplasmic domain. Like its counterparts in Drosophila, TLRs signal through adaptor molecules. The TLR family is a phylogenetically conserved mediator of innate immunity that is essential for microbial recognition. Ten human homologs of TLRs (TLR1-10) have been described. Among this family of receptors, TLR2 and TLR4 have been most studied. These studies have suggested that TLR2 and TLR4 may serve as potential main mediators of LPS signaling.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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NB100-2322
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NB100-56566
Species: Hu, Mu, Rt, Po, Bv, Ma, Pm, ChHa
Applications: WB, ChIP, DB, ELISA, Flow, Flow-CS, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, B/N, CyTOF-ready, Dual ISH-IHC, ELISA(Cap), KD, KO
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Publications for TLR4 Antibody (NB100-56566)(93)

We have publications tested in 6 confirmed species: Human, Mouse, Rat, Bovine, Mammal, Porcine.

We have publications tested in 16 applications: B/N, ChIP, DB, ELISA, FLOW, Flow, Flow-CS, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, In-vitro, KD, WB, WB,ICC/IF.


Filter By Application
B/N
(2)
ChIP
(1)
DB
(1)
ELISA
(1)
FLOW
(4)
Flow
(8)
Flow-CS
(3)
Flow-IC
(1)
ICC/IF
(10)
IHC
(7)
IHC-Fr
(1)
IHC-P
(8)
In-vitro
(1)
KD
(1)
WB
(33)
WB,ICC/IF
(1)
All Applications
Filter By Species
Human
(22)
Mouse
(19)
Rat
(26)
Bovine
(1)
Mammal
(1)
Porcine
(4)
All Species
Showing Publications 1 - 10 of 93. Show All 93 Publications.
Publications using NB100-56566 Applications Species
Bidne KL. Investigating the ovarian response to endotoxemia. Thesis Jul 9 1905 (WB, Porcine) WB Porcine
Burkovetskaya ME, Small R, Guo L et al. Cocaine self-administration differentially activates microglia in the mouse brain Neurosci. Lett. Apr 9 2020 [PMID: 32278944] (WB, Mouse) WB Mouse
Ozbek M, Hitit M, Ergun E et al. Expression profile of Toll-like receptor 4 in rat testis and epididymis throughout postnatal development Andrologia Jan 31 2020 [PMID: 32003057] (IHC, Rat) IHC Rat
Rodet F, Capuz A, Ozcan BA et al. PC1/3 KD Macrophages Exhibit Resistance to the Inhibitory Effect of IL-10 and a Higher TLR4 Activation Rate, Leading to an Anti-Tumoral Phenotype Cells Nov 22 2019 [PMID: 31766635] (ICC/IF, KD, Rat) ICC/IF, KD Rat
Evran S, Calis F, Akkaya E et al. The effect of high mobility group box-1 protein on cerebral edema, blood-brain barrier, oxidative stress and apoptosis in an experimental traumatic brain injury model Brain Res. Bull. Nov 9 2019 [PMID: 31715313] (IHC, Rat) IHC Rat
Zi SF, Li JH, Liu L et al. Dexmedetomidine-mediated protection against septic liver injury depends on TLR4/MyD88/NF-kappaB signaling downregulation partly via cholinergic anti-inflammatory mechanisms Int. Immunopharmacol. Sep 11 2019 [PMID: 31520992] (WB, Rat) WB Rat
Zhang SP, Yang RH, Shang J et al. FOXC1 up-regulates the expression of toll-like receptors in myocardial ischaemia J. Cell. Mol. Med. Sep 13 2019 [PMID: 31517441] (WB, Mouse, Human, Rat) WB Mouse, Human, Rat
Manago A, Audrito V, Mazzola F et al. Extracellular nicotinate phosphoribosyltransferase binds Toll like receptor 4 and mediates inflammation Nat Commun Sep 11 2019 [PMID: 31511522] (ICC/IF, Human) ICC/IF Human
Yang K, Lauritzen KH, Olsen MB et al. Low Cellular NAD+ Compromises Lipopolysaccharide-Induced Inflammatory Responses via Inhibiting TLR4 Signal Transduction in Human Monocytes J. Immunol. Aug 19 2019 [PMID: 31427442] (WB, Human) WB Human
Arya RP, Arankalle VA Phenotypic analysis of monocytes and CD4+ T cells in hepatitis E patients with or without pregnancy Hum. Immunol. Jul 5 2019 [PMID: 31285077] (Flow, Human) Flow Human
Show All 93 Publications.

Reviews for TLR4 Antibody (NB100-56566) (5) 4.25

Average Rating: 4.2
(Based on 5 reviews)
We have 5 reviews tested in 2 species: Mouse, Other.

Reviews using NB100-56566:
Filter by Applications
IHC
(1)
WB
(3)
All Applications
Filter by Species
Mouse
(3)
Other
(2)
All Species
Images Ratings Applications Species Date Details
 TLR4 NB100-56566
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reviewed by:
Anonymous
Mouse 05/25/2017
View

Summary

Sample Testedbrain tissue slice(30um
SpeciesMouse

Comments

CommentsIHC-frozen, 1:500
Immunohistochemistry TLR4 NB100-56566
Enlarge
reviewed by:
Hayk Simonyan
IHC Mouse 05/16/2017
View

Summary

ApplicationImmunohistochemistry
Sample Tested Mouse brain,Adult mouse brain tissue,Mouse cortex
SpeciesMouse
Western Blot TLR4 NB100-56566
Enlarge
reviewed by:
Aileen Keating
WB Other 05/12/2016
View

Summary

ApplicationWestern Blot
Sample TestedPig whole ovarian lysate
SpeciesOther
LotCTJZUO

Blocking

Blocking Details5% BSA in.2% PBST for 3 hours at room temperature, rocking

Primary Anitbody

Dilution Ratio1:500 in 5% BSA.2% PBST, overnight, at 4 degrees Celsius, rocking

Secondary Antibody

Secondary DescriptionHorse anti-mouse IgG, HRP
Secondary Manufacturer Cat#Cell Signaling: 7076
Secondary Concentration1:1000 in 5% BSA.2% PBST

Details

Detection NotesECL substrate, Alpha Innotech MultiImage Light Cabinet, 30 sec exposure. Controls negative for signal (1° only, 2° only, IgG).
Western Blot TLR4 NB100-56566
Enlarge
reviewed by:
Liyong Zhang
WB Mouse 08/07/2014
View

Summary

ApplicationWestern Blot
SpeciesMouse
FileView PDF
Western Blot TLR4 NB100-56566
Enlarge
reviewed by:
Courtney Kurtz
WB Other 08/11/2011
View

Summary

ApplicationWestern Blot
Sample Testedsmall intestinal lysate, Sample Amount: 50 ug
SpeciesOther
CommentsWorks very well with chemiluminescent detection, but not quite strong enough for a good signal with colorimetric detection. Overall good! Appears slightly smaller than normal, although this can happen in my particular species. Only band observed on blot.

Blocking

Blocking DetailsBlocking Buffer: 4% milk, Blocking Time: 1 hour, Blocking Temp: Room temperature

Primary Anitbody

Dilution RatioDilution Ratio: 1:250, Incubation Dilution Buffer: 4% milk, Incubation Time: overnight, Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab: goat anti-mouse poly-HRP, Secondary Ab Dilution Ratio: 1:1500

Details

Detection NotesDetection Method: Chemiluminescence, Exposure Time: 30 seconds, Specific Bands: ~70 kD? kDa

Comments

CommentsWorks very well with chemiluminescent detection, but not quite strong enough for a good signal with colorimetric detection. Overall good! Appears slightly smaller than normal, although this can happen in my particular species. Only band observed on blot.

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
ICC/IF Video Protocol

FAQs for TLR4 Antibody (NB100-56566). (Showing 1 - 4 of 4 FAQs).

  1. In my experiment, I need to check the TLR4 expression on Swine embryonic fibroblast cells. Is it applicable to use the human TLR4 antibody? Have you obtained any references on swine cells which using the same TLR4 antibody.
    • This antibody has not been used on swine however I have carried out a blast and the % homology is 73% between the human protein and the pig protein. If you would be interested in testing this novel species, please take a look at our Innovators Reward Program.
  2. Do you know the amino acid sequence against which this antibody is directed to? And also, what isoform of the protein would it be recognizing, or from what specific TLR4 transcript?
    • This antibody binds within amino acids 100-200 of TLR4. This antibody will recognize isoforms 1 and 2 of TLR4 as this region is present in both of these isoforms, but not isoform 3, according to UniProt O00206.
  3. Have you tested whether NB100-56566/ NB100-56728 can detect TLR4/TLR2 from NBP2-26268/NBP2-26266, respectively? If you tested, could you please inform us about the western blot procedure?
    • These cell lines are designed for flow cytometry and TLR functional assays. They are not designed for western blot analysis of TLRs. The antibodies that you mentioned do recognize the TLRs in these cells. As shown on the NBP2-26266 data sheet, the TLR2 antibody can be used in flow cytometry to recognize the TLR2 in the NBP2-26266 cell line. As shown on the NBP2-26268 data sheet, the antibody can be used in flow cytometry to recognize the TLR4 in the NBP2-26268 cell line. Please note, in general it is not uncommon that antibodies do not recognize certain recombinant constructs in one or more techniques. Our cell lines were selected for functional activity rather than for recognition by particular TLR antibodies. The construct insert was confirmed by using antibody to HA tag as shown on the data sheets.
  4. Would you be able to share the sequence homology/alignment of the immunogen sequence of NB100-56566 to the Zebrafish protein (S5Q1H4 ( uniprot site))?
    • I ran the sequence you provided against the immunogen, and unfortunately, there is only a 28% homology.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NBP2-27149AF350
Alexa Fluor 405 NBP2-27149AF405
Alexa Fluor 488 NBP2-27149AF488
Alexa Fluor 532 NBP2-27149AF532
Alexa Fluor 594 NBP2-27149AF594
Alexa Fluor 647 NBP2-27149AF647
Alexa Fluor 700 NBP2-27149AF700
Alexa Fluor 750 NBP2-27149AF750
Allophycocyanin NBP2-27149APC
Allophycocyanin/Cy7 NB100-56566APCCY7
Biotin NBP2-27149B
DyLight 350 NBP2-27149UV
DyLight 405 NBP2-27149V
DyLight 488 NBP2-27149G
DyLight 550 NBP2-27149R
DyLight 594 NB100-56566DL594
DyLight 650 NBP2-27149C
DyLight 680 NBP2-27149FR
DyLight 755 NBP2-27149IR
FITC NBP2-27149F
HRP NBP2-27149H
Janelia Fluor 549 NB100-56566JF549
Janelia Fluor 646 NB100-56566JF646
PE NBP2-27149PE
PE/Atto594 NB100-56566PEATT594
PE/Cy5.5 NB100-56566PECY55
PE/Cy7 NB100-56566PECY7
PerCP NBP2-27149PCP

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Blogs on TLR4. Showing 1-10 of 11 blog posts - Show all blog posts.

PAMPs and DAMPs: What is the same and what is different about these molecules?
By Victoria OsinskiWhat are PAMPs and DAMPsInflammation results from stimuli signaling damage or infection. The inflammatory response can be beneficial or harmful depending on the type and duration of stimuli. The s...  Read full blog post.

How to identify B cell subsets using flow cytometry
By Victoria OsinskiUsing flow cytometry to identify B cell subsetsIdentifying cellular subsets by flow cytometry requires careful and thorough planning in order to ensure the correct subset of cells are identified a...  Read full blog post.

Lipopolysaccharide from gut microbiome localizes in human atherosclerotic plaques and promotes TLR4-mediated oxidative stress
By Jamshed Arslan, Pharm. D., PhD. Atherosclerosis is a chronic inflammatory condition in which plaques of fats and other substances slowly buildup on the inner walls of arteries to restrict blood flow. In atheroscle...  Read full blog post.

Toll-like receptors in the intestinal epithelial cells
By Jamshed Arslan, Pharm. D., PhD. Toll-like receptors (TLRs) are microbe-sensing proteins that act as first responders to danger signals. TLRs help the intestinal epithelial cells (IECs) recognize commensal bacteria...  Read full blog post.

The role of STING/TMEM173 in gamma and encephalitis Herpes Simplex Virus (HSV)
Stimulator of interferon genes (STING), also known as TMEM173, promotes the production of the interferon’s IFN-alpha and IFN-beta.  STING possesses three functional domains: a cytoplasmic C-terminal tail, a central globular domain, and four N-...  Read full blog post.

TRIF/TICAM1 and mitochondrial dynamics in the innate immune response
TRIF, also known as toll like receptor adaptor molecule 1 or TICAM1, is known for its role in invading foreign pathogens as part of our innate immune response. TRIF/TICAM1 is a TIR-domain adaptor protein (toll/interleukin-1 receptor) that interacts...  Read full blog post.

The role of TLR4 in breast cancer
Toll like receptors (TLRs) are highly conserved proteins that are first known for their role in pathogen recognition and immune response activation.  In order to elicit the necessary immune response in reaction to a foreign pathogen, TLRs trigger cy...  Read full blog post.

cIAP2 - balancing cell death and cell survival
The inhibitor of apoptosis proteins (IAPs) are important regulators of cell death and inflammation. The cellular inhibitor of apoptosis protein 2 (cIAP2) contains three Baculovirus IAP repeat (BIR) domains, a Ubiquitin associated (UBA) domain, and...  Read full blog post.

TLR4 - A Guardian of Innate Immunity
Toll-like receptor 4 (TLR4) belongs to the family of Toll-like receptors (TLR), and plays a main role in pathogen recognition and innate immunity system activation. The TLR family members are highly conserved proteins that all contain a high degree of...  Read full blog post.

IRAK4: The "master IRAK" critical for initiating immune responses
IRAK4, also known as Interleukin-1 receptor-associated kinase 4, is a serine/threonine-protein kinase that plays a critical role in initiating innate and adaptive immune responses against foreign pathogens. It activates NF-kappaB in both Toll-like rec...  Read full blog post.

Showing 1-10 of 11 blog posts - Show all blog posts.
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Recent Reviews

4.2
5
1
4
4
3
0
2
0
1
0

Anonymous
05/25/2017
Application:
Species: Mouse

Hayk Simonyan
05/16/2017
Application: IHC
Species: Mouse

Aileen Keating
05/12/2016
Application: WB
Species: Other

Bioinformatics

Gene Symbol TLR4
Entrez
Uniprot