TLR7 Antibody - BSA Free

Novus Biologicals | Catalog # NBP2-24906

Novus Biologicals

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Bioactivity, Dot Blot, Proximity Ligation Assay

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Block/Neutralize, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Bioassay, Cytometric Bead Assay Standard, Flow Cytometry Control, Proximity Ligation Assay, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all TLR7 Primary Antibodies »

Product Specifications

Immunogen

Partial synthetic peptide made to an internal portion of the human TLR7 protein (between amino acids 700-750) [UniProt 9NYK1]

Reactivity Notes

Use in Human reported in scientific literature (PMID:33806288).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Novus Biologicals Rabbit TLR7 Antibody - BSA Free (NBP2-24906) is a polyclonal antibody validated for use in IHC, WB, Flow, ICC/IF, Simple Western and IP. Anti-TLR7 Antibody: Cited in 81 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for TLR7 Antibody - BSA Free

Immunohistochemistry: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry: TLR7 Antibody - BSA Free [NBP2-24906]

TLR7-Antibody-Immunohistochemistry-NBP2-24906-img0022.jpg
Flow Cytometry: TLR7 Antibody - BSA Free [NBP2-24906]

Flow Cytometry: TLR7 Antibody - BSA Free [NBP2-24906]

Flow Cytometry: TLR7 Antibody [NBP2-24906] - An intracellular stain was performed on THP-1 cells with NBP2-24906 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).
Immunocytochemistry/ Immunofluorescence: TLR7 Antibody - BSA Free [NBP2-24906]

Immunocytochemistry/ Immunofluorescence: TLR7 Antibody - BSA Free [NBP2-24906]

Immunocytochemistry/Immunofluorescence: TLR7 Antibody [NBP2-24906] - Immunolocalization of TLR7 (A) and glial fibrillary acidic protein (GFAP) (B) in murine myenteric plexus. Most of the GFAP positive cells were also positive for TLR7 in the merged image (C, yellow staining). (Courtesy of Barajon I, et al., Journal of Histochemistry and Cytochemistry, 57(11): 1013-;1023, 2009.)
Immunohistochemistry: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry: TLR7 Antibody - BSA Free [NBP2-24906]

TLR7-Antibody-Immunohistochemistry-NBP2-24906-img0023.jpg
Immunohistochemistry-Frozen: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry-Frozen: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry-Frozen: TLR7 Antibody [NBP2-24906] - analysis of frozen mouse ear skin tissue using anti-TLR7 antibody. Image from verified customer review.
Immunohistochemistry-Frozen: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry-Frozen: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry-Frozen: TLR7 Antibody [NBP2-24906] - analysis of TLR7 in acetone-fixed, frozen mouse ear skin section using anti-TLR7 antibody. Image from verified customer review.
Western Blot: TLR7 AntibodyBSA Free [NBP2-24906]

Western Blot: TLR7 AntibodyBSA Free [NBP2-24906]

Western Blot: TLR7 Antibody [NBP2-24906] - Analysis using the Azide Free version of NBP2-24906. Detection of TLR7 in RAW cell lysate using this antibody.
Immunocytochemistry/ Immunofluorescence: TLR7 Antibody - BSA Free [NBP2-24906]

Immunocytochemistry/ Immunofluorescence: TLR7 Antibody - BSA Free [NBP2-24906]

Immunocytochemistry/Immunofluorescence: TLR7 Antibody [NBP2-24906] - Analysis using the Azide Free version of NBP2-24906. Staining of Raw 246.7 cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Image objective 40x. An antibody dilution of 1:10 was used.
Immunohistochemistry-Paraffin: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry-Paraffin: TLR7 Antibody - BSA Free [NBP2-24906]

Immunohistochemistry-Paraffin: TLR7 Antibody - BSA Free [NBP2-24906] - Analysis of a FFPE tissue section of human skin using 1:200 dilution of TLR7 antibody (NBP2-24906). The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin.
TLR7 Antibody - BSA Free

Immunohistochemistry-Paraffin: TLR7 Antibody - BSA Free [NBP2-24906] -

Immunohistochemistry-Paraffin: TLR7 Antibody - BSA Free [NBP2-24906] - Analysis of human colon tissue using NBP2-24905 (top) and an isotype control (bottom) at 5 ug/ml.
TLR7 Antibody - BSA Free

Immunohistochemistry: TLR7 Antibody - BSA Free [NBP2-24906] -

Immunohistochemistry: TLR7 Antibody - BSA Free [NBP2-24906] - Expression of TLR7 in lung tissues of non-smokers, smokers, & COPD patients. Total protein & mRNA were obtained from lung tissues of non-smokers (n = 15), smokers (n = 12), & COPD patients (n = 15). TLR7 protein & mRNA expression were determined by western blot (A) & real time PCR (B) in lung parenchyma. (A) Representative images of western blot for TLR7 & corresponding densitometry expressed as ratio to beta -actin. (B) TLR7 mRNA expression given as the ratio to GAPDH. (C, D, E) Lung sections were immunostained for TLR7 & quantified by means of immunohistochemical score of TLR7 in alveolar macrophages (C) & bronchial epithelial cells (D). (E) Representative immunohistochemistry images are shown. The control IgG isotype showed negative staining. Data are presented as individual values & mean ± standard deviation. Exact P values were obtained using Kruskal-Wallis & Dunn’s post-hoc tests. Image collected & cropped by CiteAb from the following publication (https://respiratory-research.biomedcentral.com/articles/10.1186/s12931-…), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for TLR7 Antibody - BSA Free

Application
Recommended Usage

Bioactivity

reported in scientific literature (PMID 28894449)

Dot Blot

reported in scientific literature (PMID 27248820)

Flow Cytometry

2-5 ug/ 1x10^6 cells

Immunocytochemistry/ Immunofluorescence

1:10-1:2000

Immunohistochemistry

1:200. Use reported in scientific literature (PMID 28219705)

Immunohistochemistry-Frozen

1:10-1:2000. Use reported by customer review

Immunohistochemistry-Paraffin

1:200. Use reported in scientific literature (PMID 25264223)

Immunoprecipitation

reported in scientific literature (PMID 17452530)

Proximity Ligation Assay

reported in scientific literature (PMID 30579042)

Simple Western

1:20

Western Blot

1-3 ug/ml

Reviewed Applications

Read 4 reviews rated 5 using NBP2-24906 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

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: TLR7

Toll-like receptor 7 (TLR7) is a type I transmembrane protein expressed on the surface of endosomes and has a role in pathogen-associated molecular patterns (PAMPs) recognition and host defense (1-3). TLR7 is primarily expressed in the brain, placenta, spleen, stomach, and lungs (4). TLR7 recognizes microbial single stranded RNA (ssRNA), specifically guanosine and its derivatives (1-3). Human TLR7 cDNA encodes a 1049 amino acid (aa) protein with a theoretical molecular weight (MW) of 120.9 kDa (4). The TLR7 protein consists of a signal sequence, an 813 aa extracellular domain containing leucine-rich repeats (LRRs) which form a horseshoe-like shape, a 21 aa transmembrane domain, and a 189 aa cytoplasmic domain with cytosolic Toll-interleukin-1 receptor homology (TIR) domains (1,2,4). TLR7 and its fellow subfamily members, TLR8 and TLR9, possess a characteristic Z-loop between two LRRs with proteolytic Z-loop processing required for TLR activation (2). Z-loop cleavage in TLR7 allows for guanosine and uridine-rich ssRNA binding to the 1st and 2nd ligand binding site, respectively (2). The TIR domain associates with the adaptor protein myeloid differentiation primary response protein (MyD88) to initiate downstream signaling (1-3,5,6). Following activation by PAMPs, TLR7 dimerizes and bound MyD88 interacts with interleukin-1 receptor-associated kinase-4 (IRAK-4) (1,5). Together the complex recruits IRAK-1 and IRAK-2, which become phosphorylated, and interact with tumor necrosis factor receptor-associated factor 6 (TRAF6) (1,5). TRAF6 induces the activation of mitogen-activated protein kinase (MAPK), nuclear factor-kappaB (NF-kappaB), and interferon-regulatory factor 7 (IRF7), leading to interferon production and pro-inflammatory cytokine secretion associated with immune response (1,5).

While TLRs play an important role in innate immune response, dysfunction in the TLR-MyD88 signaling cascade has also been reported in various autoimmune disorders (5,6). Elevated expression of TLR7 is associated with increased risk of system lupus erythematosus (SLE), an autoimmune disease involving B cell hyperactivity (6,7). Studies involving mouse models has also found that increased TLR7 expression predisposes mice to a lupus-like disease (7). Therapeutics targeting TLR7 have been developed to either enhance or inhibit its activity depending on the circumstance. For example, TLR7 agonists such as imiquimod, resiquimod, and 852A are used to increase TLR7 activity for treatment of cancers and to fight viral infections (7,8). On the other hand, TLR7 antagonists inhibit its activation and have been developed to combat chronic immune stimulation as seen in inflammatory and autoimmune diseases (8).

References

1. Petes C, Odoardi N, Gee K. The Toll for Trafficking: Toll-Like Receptor 7 Delivery to the Endosome. Front Immunol. 2017;8:1075. https://doi.org/10.3389/fimmu.2017.01075

2. Maeda K, Akira S. TLR7 Structure: Cut in Z-Loop. Immunity. 2016;45(4):705-707. https://doi.org/10.1016/j.immuni.2016.10.003

3. Krieg AM, Vollmer J. Toll-like receptors 7, 8, and 9: linking innate immunity to autoimmunity. Immunol Rev. 2007;220:251-269. https://doi.org/10.1111/j.1600-065X.2007.00572.x

4. Uniprot (Q9NYK1)

5. Zheng C, Chen J, Chu F, Zhu J, Jin T. Inflammatory Role of TLR-MyD88 Signaling in Multiple Sclerosis. Front Mol Neurosci. 2020;12:314. https://doi.org/10.3389/fnmol.2019.00314

6. Chi H, Li C, Zhao FS, et al. Anti-tumor Activity of Toll-Like Receptor 7 Agonists. Front Pharmacol. 2017;8:304. https://doi.org/10.3389/fphar.2017.00304

7. Fillatreau S, Manfroi B, Dorner T. Toll-like receptor signalling in B cells during systemic lupus erythematosus. Nat Rev Rheumatol. 2021;17(2):98-108. https://doi.org/10.1038/s41584-020-00544-4

8. Patinote C, Karroum NB, Moarbess G, et al. Agonist and antagonist ligands of toll-like receptors 7 and 8: Ingenious tools for therapeutic purposes. Eur J Med Chem. 2020;193:112238. https://doi.org/10.1016/j.ejmech.2020.112238

Long Name

Toll-like Receptor 7

Alternate Names

toll-like receptor 7

Gene Symbol

TLR7

Additional TLR7 Products

Product Documents for TLR7 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for TLR7 Antibody - 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 TLR7 Antibody - BSA Free

Customer Reviews for TLR7 Antibody - BSA Free (4)

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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • TLR7 Antibody
    Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Fixe mouse brain tissue
    Species: Mouse
    Verified Customer | Posted 08/02/2019
    Immunostaing of 4% PFA fixed mouse brain tissue, stained with Rabbi anti-TLR7 (dilution 1:500; Alexa 488 green).
    TLR7 Antibody - BSA Free NBP2-24906
  • TLR7 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Dorsal root ganglion neurons
    Species: Rat
    Verified Customer | Posted 12/02/2017
    TLR7 Antibody - BSA Free NBP2-24906
  • Name: Anonymous
    Application: Immunohistochemistry-Frozen
    Sample Tested: Mouse Ear Skin (frozen section)
    Species: Mouse
    Verified Customer | Posted 08/29/2015
    Mouse Ear Skin (frozen section)
    TLR7 Antibody - BSA Free NBP2-24906
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Mouse Ear Skin (frozen section)
    Species: Mouse
    Verified Customer | Posted 08/28/2015
    Mouse Ear Skin
    TLR7 Antibody - BSA Free NBP2-24906

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Protocols

View specific protocols for TLR7 Antibody - BSA Free (NBP2-24906):

Protocol for Flow Cytometry Intracellular Staining
Sample Preparation.
1. Grow cells to 60-85% confluency. Flow cytometry requires between 2 x 105 and 1 x 106 cells for optimal performance.
2. If cells are adherent, harvest gently by washing once with staining buffer and then scraping. Avoid using trypsin as this can disrupt certain epitopes of interest. If enzymatic harvest is required, use Accutase, Collagenase, or TrypLE Express for a less damaging option.
3. Reserve 100 uL for counting, then transfer cell volume into a 50 mL conical tube and centrifuge for 8 minutes at 400 RCF.
a. Count cells using a hemocytometer and a 1:1 trypan blue exclusion stain to determine cell viability before starting the flow protocol. If cells appear blue, do not proceed.
4. Re-suspend cells to a concentration of 1 x 106 cells/mL in staining buffer (NBP2-26247).
5. Aliquot out 100 uL samples in accordance with your experimental samples.

Tip: When cell surface and intracellular staining are required in the same sample, it is advisable that the cell surface staining be performed first since the fixation and permeabilization steps might reduce the availability of surface antigens.

Intracellular Staining.
Tip: When performing intracellular staining, it is important to use appropriate fixation and permeabilization reagents based upon the target and its subcellular location. Generally, our Intracellular Flow Assay Kit (NBP2-29450) is a good place to start as it contains an optimized combination of reagents for intracellular staining as well as an inhibitor of intracellular protein transport (necessary if staining secreted proteins). Certain targets may require more gentle or transient permeabilization protocols such as the commonly employed methanol or saponin-based methods.
Protocol for Cytoplasmic Targets:
1. Fix the cells by adding 100 uL fixation solution (such as 4% PFA) to each sample for 10-15 minutes.
2. Permeabilize cells by adding 100 uL of a permeabilization buffer to every 1 x 106 cells present in the sample. Mix well and incubate at room temperature for 15 minutes.
a. For cytoplasmic targets, use a gentle permeabilization solution such as 1X PBS + 0.5% Saponin or 1X PBS + 0.5% Tween-20.
b. To maintain the permeabilized state throughout your experiment, use staining buffer + 0.1% of the permeabilization reagent (i.e. 0.1% Tween-20 or 0.1% Saponin).
3. Following the 15 minute incubation, add 2 mL of the staining buffer + 0.1% permeabilizer to each sample.
4. Centrifuge for 1 minute at 400 RCF.
5. Discard supernatant and re-suspend in 100 uL of staining buffer + 0.1% permeabilizer.
6. Add appropriate amount of each antibody (eg. 1 test or 1 ug per sample, as experimentally determined).
7. Mix well and incubate at room temperature for 30 minutes- 1 hour. Gently mix samples every 10-15 minutes.
8. Following the primary/conjugate incubation, add 1-2 mL/sample of staining buffer +0.1% permeabilizer and centrifuge for 1 minute at 400 RCF.
9. Wash twice by re-suspending cells in staining buffer (2 mL for tubes or 200 uL for wells) and centrifuging at 400 RCF for 5 minutes. Discard supernatant.
10. Add appropriate amount of secondary antibody (as experimentally determined) to each sample.
11. Incubate at room temperature in dark for 20 minutes.
12. Add 1-2 mL of staining buffer and centrifuge at 400 RCF for 1 minute and discard supernatant.
13. Wash twice by re-suspending cells in staining buffer (2 mL for tubes or 200 uL for wells) and centrifuging at 400 RCF for 5 minutes. Discard supernatant.
14. Resuspend in an appropriate volume of staining buffer (usually 500 uL per sample) and proceed with analysis on your flow cytometer.

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 4% paraformaldehyde to the dish and fix at room temperature for 10 minutes.
2. Remove the paraformaldehyde and wash the cells in PBS.
3. Permeabilize the cells with 0.1% Triton X100 or other suitable detergent for 2 min.
4. Remove the permeabilization buffer and wash three times for 5 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 5 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 5 minutes each.
10. Counter stain DNA with DAPI if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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FAQs for TLR7 Antibody - BSA Free

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    • Q: Regarding antibody NBP2-24906 is the western blot of RAW264.7 cells detected with colorimetric staining? or is it luminescence (ECL)?
      A: Image 5 of 13 is ECL
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