TLR4 Antibody (76B357.1) - Azide and BSA Free

Novus Biologicals | Catalog # NBP2-27149

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Mammal

Cited:

Human, Mouse, Rat

Predicted:

Baboon (100%), Chimpanzee (100%), Chinese Hamster (100%). Backed by our 100% Guarantee.

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Block/Neutralize, Flow Cytometry, Flow (Cell Surface), Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Knockdown Validated, CyTOF-ready

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Block/Neutralize, Flow Cytometry, Flow (Cell Surface), Immunocytochemistry/ Immunofluorescence, Flow Cytometry Control

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2b Kappa Clone # 76B357.1

Format

Azide and BSA Free
View all TLR4 Primary Antibodies »

Product Specifications

Immunogen

This TLR4 Antibody (76B357.1) was developed against a portion of amino acids 100-200 of human TLR4 (NP_612564).

Reactivity Notes

Ground squirrel reactivity reported by a verified customer review. Mammal reactivity reported in scientific literature (PMID: 25130694).

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2b Kappa

Theoretical MW

95.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for TLR4 Antibody (76B357.1) - Azide and BSA Free

Western Blot: TLR4 Antibody (76B357.1)Azide Free [NBP2-27149]

Western Blot: TLR4 Antibody (76B357.1)Azide Free [NBP2-27149]

Western Blot: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Analysis of TLR4 using TLR4 antibody at 2 ug/mL on (A) human intestine and 6 ug/mL on (B) mouse intestine and C) rat intestine lysate.
Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Tissue section of normal human skin stained with antibody at 5 ug/mL. Membrane-cytoplasmic immunopositivity of TLR4 was primarily observed in the pigmented basel cells and the adjacent keratinocytes in the epidermal layer. Image using the standard format of this product.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - 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 anti-mouse IgG (H+L) cross adsorbed secondary antibody, (DyLight 488 conjugated). Isotype control samples incubated with mouse IgG2b isotype control antibody were processed in parallal under the same assay conditions. Image using the standard format of this product.
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - FFPE human testis tissue section stained with TLR4 antibody at 5 ug/mL.
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Human skin stained with TLR4 antibody at 5 ug/mL, peroxidase-conjugate and DAB chromogen.
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Formalin-fixed paraffin-embedded tissue sections of human tonsil were probed for TLR4 mRNA (ACD RNAScope Probe, catalog # 311281; Fast Red chromogen, ACD catalog # 322750). Adjacent tissue section was processed for immunohistochemistry using Mouse Monoclonal (Novus Biologicals catalog # NB100-56566) at 5ug/mL with 1 hour incubation at room temperature followed by incubation with anti-mouse IgG VisUCyte HRP Polymer Antibody (Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. Image using the standard format of this product.
Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Immunofluorescent staining of TMAs. Representative tissue cores from normal (I), adenomatous polyps (II), and CRC (III and IV) are shown. Image collected and cropped by CiteAb from the following publication (https://www.jeccr.com/content/33/1/45), licensed under a CC-BY license. Image using the standard format of this product.
Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. Double-stained immunofluorescence for TLR4 (green) and vimentin (red) in normal (I), adenoma (II), and colon adenocarcinoma (III) (10x). In the stromal compartment of CRCs, immunofluorescent staining for TLR4 localized to the pericryptal myofibroblasts in a subset of samples. Image collected and cropped by CiteAb from the following publication (https://www.jeccr.com/content/33/1/45), licensed under a CC-BY license. Image using the standard format of this product.
Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Pericryptal Myofibroblasts are Responsible for Increased TLR4 Expression in a Subset of CRCs. IHC staining of colon adenocarcinoma for TLR4, vimentin, and alpha-SMA (40x). Staining co-localizes to the pericryptal space, confirming the signal arises from pericryptal myofibroblasts. Image collected and cropped by CiteAb from the following publication (https://www.jeccr.com/content/33/1/45), licensed under a CC-BY license. Image using the standard format of this product.
Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - This image shows TLR4 (green) and nucleus (blue) of area postrema of the adult male mouse brain, 20X magnification. Primary antibody diluted 1:500. IHC image submitted by a verified customer review.
Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Analysis of Rat's salivary gland tissue section at 1:100 dilution. The antibody generated a membrane-cytoplasmic staining in the tissue with stronger signal in ductal epithelial cells. Image using the standard format of this product.
Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Immunohistochemistry-Paraffin: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Analysis of TLR4 in paraffin-embedded formalin-fixed human colon tissue using an isotype control (top) and NB100-56566 (bottom) at 5 ug/mL. Image using the standard format of this product.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Analysis of cell surface TLR4 in mouse splenocytes using 0.5 ug/10^6 cells of TLR4 antibody. Shaded histogram represents cells without TLR4 antibody; green represents isotype control (BD Pharmingen) red represents TLR4 antibody.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - 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 test, with CD14 PE this antibody and 1 ug of either isotype control (Left, ) or TLR4-Alexa Fluor 647 (right). PPI negative, CD14+ cells were gated for analysis.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Analysis using the FITC conjugate of NBP2-27149. TLR4 expression on monocytes from human peripheral blood: PBMC were stained, in a 2 color flow test, with CD14 PE and 1 ug of either isotype control (Left, ) or TLR4-FITC (Right, this antibody. PPI negative, CD14+ cells were gated for analysis.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - Analysis using the FITC conjugate of NBP2-27149. Staining of cell surface TLR4 in human monocyte-derived dendritic cells using this antibody at 1:200 (2.5 ug). Black and green open histograms represent unstained cells and negative control (anti-human CD14).
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - 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 isotype control MAB004 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Allophycocyanin.
Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149]

Flow Cytometry: TLR4 Antibody (76B357.1) - Azide Free [NBP2-27149] - An intracellular stain was performed on Jurkat cells with TLR4 antibody (76B357.1) NBP2-27149AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Applications for TLR4 Antibody (76B357.1) - Azide and BSA Free

Application
Recommended Usage

Block/Neutralize

reported in scientific literature (PMID 26063617)

Chromatin Immunoprecipitation (ChIP)

1:10-1:500

Flow (Cell Surface)

reported in multiple pieces of scientific literature

Flow (Intracellular)

reported in scientific literature (PMID 18577706)

Flow Cytometry

1ul/1 million cells

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry-Frozen

1:500

Immunohistochemistry-Paraffin

5ug/ml

Western Blot

1-3 ug/ml

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: TLR4

TLR4 (Toll-like receptor 4) is a type-1 transmembrane glycoprotein that is a pattern recognition receptor (PRR) belonging to the TLR family (1-3). TLR4 is expressed in many tissues and is most abundantly expressed in the placenta, spleen, and peripheral blood leukocytes (1). Human TLR4 is synthesized as a 839 amino acid (aa) protein containing a signal sequence (1-23 aa), an extracellular domain (ECD) (24-631 aa), a transmembrane domain (632-652 aa), and Toll/interleukin-1 receptor (TIR) cytoplasmic domain (652-839 aa) with a theoretical molecular weight of 95 kDa (3, 4). The ECD contains 21 leucine-rich repeats (LRRs) and has a horseshoe-shaped structure (3, 4). TLR4 requires binding with the co-receptor myeloid differentiation protein 2 (MD2) largely via hydrophilic interactions for proper ligand sensing and signaling (2-4). In general, the TLR family plays a role in activation of innate immunity and responds to a variety of pathogen-associated molecular patterns (PAMPs) (5). TLR4 is specifically responsive to lipopolysaccharide (LPS), which is found on the outer-membrane of most ram-negative bacteria (3-5). Activation of TLR4 requires binding of a ligand, such as LPS to MD2, followed by MD2-LPS complex binding to TLR4, resulting in a partial complex (TLR4-MD2/LPS) (3, 5). To become fully active, two partial complexes must dimerize thereby allowing the TIR domains of TLR4 to bind other adapter molecular and initiate signaling, triggering an inflammatory response and cytokine production (3, 5).

TLR4 signaling occurs through two distinct pathways: The MyD88 (myeloid differentiation primary response gene 88)-dependent pathway and the MyD88-independent (TRIF-dependent, TIR domain-containing adaptor inducing IFN-beta) pathway (3, 5-7). The MyD88-dependent pathway occurs mainly at the plasma membrane and involves the binding of MyD88-adaptor-like (MAL) protein followed by a signaling cascade that results in the activation of transcription factors including nuclear factor-kappaB (NF-kappaB) that promote the secretion of inflammatory molecules and increased phagocytosis (5-7). Conversely, the MyD88-independent pathway occurs after TLR4-MD2 complex internalization in the endosomal compartment. This pathway involves the binding of adapter proteins TRIF and TRIF-related adaptor molecule (TRAM), a signaling activation cascade resulting in IFN regulatory factor 3 (IRF3) translocation into the nucleus, and secretion of interferon-beta (INF-beta) genes and increased phagocytosis (5-7).

Given its expression on immune-related cells and its role in inflammation, TLR4 activation can contribute to various diseases (6-8). For instance, several studies have found that TLR4 activation is associated with neurodegeneration and several central nervous system (CNS) pathologies, including Alzheimer's disease, Parkinson's disease, and Huntington's disease (6, 7). Furthermore, TLR4 mutations have been shown to lead to higher rates of infections and increased susceptibility to sepsis (7-8). One potential therapeutic approach aimed at targeting TLR4 and neuroinflammation is polyphenolic compounds which include flavonoids and phenolic acids and alcohols (8).

Alternative names for TLR4 includes 76B357.1, ARMD10, CD284 antigen, CD284, EC 3.2.2.6, homolog of Drosophila toll, hToll, toll like receptor 4 protein, TOLL, toll-like receptor 4.

References

1. Vaure, C., & Liu, Y. (2014). A comparative review of toll-like receptor 4 expression and functionality in different animal species. Frontiers in immunology. https://doi.org/10.3389/fimmu.2014.00316

2. Park, B. S., & Lee, J. O. (2013). Recognition of lipopolysaccharide pattern by TLR4 complexes. Experimental & molecular medicine. https://doi.org/10.1038/emm.2013.97

3. Krishnan, J., Anwar, M.A., & Choi, S. (2016) TLR4 (Toll-Like Receptor 4). In: Choi S. (eds) Encyclopedia of Signaling Molecules. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-6438-9_592-1

4. Botos, I., Segal, D. M., & Davies, D. R. (2011). The structural biology of Toll-like receptors. Structure. https://doi.org/10.1016/j.str.2011.02.004

5. Lu, Y. C., Yeh, W. C., & Ohashi, P. S. (2008). LPS/TLR4 signal transduction pathway. Cytokine. https://doi.org/10.1016/j.cyto.2008.01.006

6. Leitner, G. R., Wenzel, T. J., Marshall, N., Gates, E. J., & Klegeris, A. (2019). Targeting toll-like receptor 4 to modulate neuroinflammation in central nervous system disorders. Expert opinion on therapeutic targets. https://doi.org/10.1080/14728222.2019.1676416

7. Molteni, M., Gemma, S., & Rossetti, C. (2016). The Role of Toll-Like Receptor 4 in Infectious and Noninfectious Inflammation. Mediators of inflammation. https://doi.org/10.1155/2016/6978936

8. Rahimifard, M., Maqbool, F., Moeini-Nodeh, S., Niaz, K., Abdollahi, M., Braidy, N., Nabavi, S. M., & Nabavi, S. F. (2017). Targeting the TLR4 signaling pathway by polyphenols: A novel therapeutic strategy for neuroinflammation. Ageing research reviews. https://doi.org/10.1016/j.arr.2017.02.004

Long Name

Toll-like Receptor 4

Alternate Names

CD284

Gene Symbol

TLR4

Additional TLR4 Products

Product Documents for TLR4 Antibody (76B357.1) - Azide and BSA Free

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Product Specific Notices for TLR4 Antibody (76B357.1) - Azide and BSA Free

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.

Citations for TLR4 Antibody (76B357.1) - Azide and BSA Free

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for TLR4 Antibody (76B357.1) - Azide and BSA Free

Showing  1 - 2 of 2 FAQs Showing All

  • Q: How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?

    A: Use isotype-specific secondary antibodies if the primary antibodies are of different isotypes. You can also make direct conjugates of the primary antibodies by use of antibody labeling kits, dyes, or custom conjugations (please contact Technical Support for custom orders).

  • Q: I would like to use this antibody but it has not been validated in my species of interest. Is there any way I can find out if it will work?

    A: We offer risk-free testing of all of our primary antibodies. Please check out our Innovator's Reward Program and test this TLR4 antibody in any unvalidated species or application, without the financial risk of failure.

  • Q: How do I choose secondary antibodies to label the same cells when I have two primary antibodies from the same host?

    A: Use isotype-specific secondary antibodies if the primary antibodies are of different isotypes. You can also make direct conjugates of the primary antibodies by use of antibody labeling kits, dyes, or custom conjugations (please contact Technical Support for custom orders).

  • Q: I would like to use this antibody but it has not been validated in my species of interest. Is there any way I can find out if it will work?

    A: We offer risk-free testing of all of our primary antibodies. Please check out our Innovator's Reward Program and test this TLR4 antibody in any unvalidated species or application, without the financial risk of failure.

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