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Product Info Summary
| SKU: | PB9451 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, ICC, WB |
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Product info
Product Name
Anti-TLR5 Antibody Picoband®
SKU/Catalog Number
PB9451
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-TLR5 Antibody Picoband® catalog # PB9451. Tested in Flow Cytometry, IHC, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-TLR5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9451)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human TLR5, different from the related mouse sequence by five amino acids.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9451 is reactive to TLR5 in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
98 kDa
Calculated molecular weight
97834 MW
Background of TLR5
TLR5 encodes a member of the toll-like receptor (TLR) family, which plays a fundamental role in pathogen recognition and activation of innate immune responses. These receptors recognize distinct pathogen-associated molecular patterns that are expressed on infectious agents. The protein encoded by this gene recognizes bacterial flagellin, the principal component of bacterial flagella and a virulence factor. The activation of this receptor mobilizes the nuclear factor NF-kappaB, which in turn activates a host of inflammatory-related target genes. Mutations in this gene have been associated with both resistance and susceptibility to systemic lupus erythematosus, and susceptibility to Legionnaire disease.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
PB9451 is guaranteed for Flow Cytometry, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: JURKAT Whole Cell,
IHC: human appendicitis tissue
FCM: Human PBMC cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of TLR5 using anti-TLR5 antibody (PB9451).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: JURKAT Whole Cell Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-TLR5 antigen affinity purified polyclonal antibody (Catalog # PB9451) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for TLR5 at approximately 98KD. The expected band size for TLR5 is at 83KD.
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Figure 2. IHC analysis of TLR5 using anti-TLR5 antibody (PB9451).
TLR5 was detected in paraffin-embedded section of human appendicitis tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TLR5 Antibody (PB9451) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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Figure 3. Flow Cytometry analysis of H-PBMC cells using anti-TLR5 antibody (PB9451).
Overlay histogram showing H-PBMC cells stained with PB9451 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-TLR5 Antibody (PB9451,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For TLR5 (Source: Uniprot.org, NCBI)
Gene Name
TLR5
Full Name
Toll-like receptor 5
Weight
97834 MW
Superfamily
Toll-like receptor family
Alternative Names
Toll-like receptor 5;Toll/interleukin-1 receptor-like protein 3;TLR5;TIL3; TLR5 MELIOS, SLE1, SLEB1, TIL3 toll like receptor 5 toll-like receptor 5|systemic lupus erythematosus susceptibility 1|toll/interleukin-1 receptor-like protein 3
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on TLR5, check out the TLR5 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for TLR5: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-TLR5 Antibody Picoband® (PB9451)
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Customer Q&As
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6 Customer Q&As for Anti-TLR5 Antibody Picoband®
Question
We are currently using anti-TLR5 antibody PB9451 for human tissue, and we are content with the WB results. The species of reactivity given in the datasheet says human. Is it possible that the antibody can work on feline tissues as well?
J. Zhao
Verified customer
Asked: 2020-04-15
Answer
The anti-TLR5 antibody (PB9451) has not been tested for cross reactivity specifically with feline tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in feline you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-04-15
Question
My question regarding product PB9451, anti-TLR5 antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2020-02-21
Answer
We suggest not storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free PB9451 anti-TLR5 antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2020-02-21
Question
Will anti-TLR5 antibody PB9451 work for IHC with cns?
Verified Customer
Verified customer
Asked: 2019-08-12
Answer
According to the expression profile of cns, TLR5 is highly expressed in cns. So, it is likely that anti-TLR5 antibody PB9451 will work for IHC with cns.
Boster Scientific Support
Answered: 2019-08-12
Question
Here is the WB image, lot number and protocol we used for cns using anti-TLR5 antibody PB9451. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2018-05-03
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-05-03
Question
Would PB9451 anti-TLR5 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2017-10-27
Answer
It shows on the product datasheet, PB9451 anti-TLR5 antibody as been tested on IHC. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2017-10-27
Question
Is this PB9451 anti-TLR5 antibody reactive to the isotypes of TLR5?
B. Zhang
Verified customer
Asked: 2015-06-04
Answer
The immunogen of PB9451 anti-TLR5 antibody is A synthetic peptide corresponding to a sequence at the C-terminus of human TLR5 (801-828aa MKHQSIRGFVQKQQYLRWPEDFQDVGWF), different from the related mouse sequence by five amino acids. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2015-06-04
