Q: I am a bit unsure about your antibody NBP2-24729 specific against TLR9. I used it for probing mouse B cell lysate on Proteisimple WES. On their website the recommend your antibody and claim that it detects a specific signal at 90 kDa. But on your website simple western detects a band at 200 kDa?

A: This came during the race to 1000 and we had many people generating and reviewing data. In retrospect, this was one that probably should not have made it out. As I reviewed the run data, it looks as though the MW ladder did not assign correctly. When this was corrected, the band at 200 actually shifts higher, up to 280 kDa. This band now overlaps with the 280 kDa fluorescent standard which is prone to have non-specific interactions with some antibodies. Those two things make this non-supportive in Simple Western. We ran Raji lysate on the same run. This antibody detects a very broad band with a peak at ~150-165 kDa, closer to expected MW. However, the baseline for this samples was also very high. If we were presented with this data set today, it would not be considered supportive. The other information on the Protein Simple web site likely came from a customer.

TLR9 Antibody (26C593.2) - BSA Free

Novus Biologicals | Catalog # NBP2-24729

(4)

Citations (166)

Product Documents

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Product Specifications
Scientific Data
Applications
Flow Cytometry
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat, Canine, Equine, Monkey, Primate

Cited:

Human, Mouse, Rat, Porcine, Canine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Block/Neutralize, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Dot Blot, Functional, In vitro assay, Knockdown Validated, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Block/Neutralize, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Cytometric Bead Assay Standard, Flow Cytometry Control, In vitro assay, Functional Assay, IF/IHC, Knockdown Validated

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ kappa Clone # 26C593.2

Format

BSA Free

View all TLR9 Primary Antibodies »

Product Specifications

Immunogen

This antibody was developed against KLH-conjugated synthetic peptide corresponding to amino acids 268-300 of human TLR9 isoform A (Genbank accession no. AAF78037).

Reactivity Notes

Rhesus Monkey.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁ kappa

Description

Novus Biologicals Mouse TLR9 Antibody (26C593.2) - BSA Free (NBP2-24729) is a monoclonal antibody validated for use in IHC, WB, ELISA, Flow, ICC/IF, Simple Western and IP. Anti-TLR9 Antibody: Cited in 152 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for TLR9 Antibody (26C593.2) - BSA Free

Simple Western: TLR9 Antibody (26C593.2)BSA Free [NBP2-24729]

Simple Western: TLR9 Antibody (26C593.2) [NBP2-24729] - Lane view shows a specific band for TLR9 in 0.5 mg/ml of Ramos lysate. This experiment was performed under reducing conditions using the 66-440 kDa separation system. Image using the Azide Free format of this antibody.

Flow Cytometry: TLR9 Antibody (26C593.2) - BSA Free [NBP2-24729]

TLR9-Antibody-26C593-2-Flow-Cytometry-NBP2-24729-img0025.jpg

Western Blot: TLR9 Antibody (26C593.2) - BSA Free [NBP2-24729]

TLR9-Antibody-26C593-2-Knockdown-Validated-NBP2-24729-img0031.jpg

Flow Cytometry: TLR9 Antibody (26C593.2) - BSA Free [NBP2-24729]

TLR9-Antibody-26C593-2-Flow-Cytometry-NBP2-24729-img0027.jpg

Immunocytochemistry/ Immunofluorescence: TLR9 Antibody (26C593.2) - BSA Free [NBP2-24729]

TLR9-Antibody-26C593-2-Immunocytochemistry-Immunofluorescence-NBP2-24729-img0032.jpg

Immunohistochemistry-Paraffin: TLR9 Antibody (26C593.2) - BSA Free [NBP2-24729]

Immunohistochemistry-Paraffin: TLR9 Antibody (26C593.2) [NBP2-24729] - Monkey retina tissue. Image from verified customer review.

Flow Cytometry: TLR9 Antibody (26C593.2) - BSA Free [NBP2-24729]

TLR9-Antibody-26C593-2-Flow-Cytometry-NBP2-24729-img0028.jpg

Western Blot: TLR9 Antibody (26C593.2)BSA Free [NBP2-24729]

Western Blot: TLR9 Antibody (26C593.2) [NBP2-24729] - Analysis of TLR9 in A) human PBMC, B) human intestine, C) mouse intestine, and D) rat intestine tissue lysates using this antibody at a dilution of 3 ug/ml.

Flow Cytometry: TLR9 Antibody (26C593.2) - BSA Free [NBP2-24729]

TLR9-Antibody-26C593-2-Flow-Cytometry-NBP2-24729-img0030.jpg

Detection of TLR9 (26C593.2) in THP-1 Acute Monocytic Leukemia Human Cell Line by Flow Cytometry.

THP-1 human acute monocytic leukemia cell line was stained with Mouse anti-TLR9 (26C593.2) Protein-G purified Monoclonal Antibody conjugated to Janelia Fluor® 549 (Catalog # NBP2-24729JF549, blue histogram) or matched control antibody (orange histogram).

View 7 more images

Applications for TLR9 Antibody (26C593.2) - BSA Free

Application

Recommended Usage

Block/Neutralize

reported in scientific literature (PMID 25338738)

Dot Blot

reported in scientific literature (PMID 27248820)

ELISA

1:100 - 1:2000. Use reported in multiple pieces of scientific literature

Flow (Intracellular)

1:10 - 1:1000. Use reported in scientific literature (PMID 24986635)

Functional

reported in scientific literature (PMID 25411258)

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

reported in scientific literature (PMID 27744078)

Immunohistochemistry-Paraffin

5 ug/ml

Immunoprecipitation

1:10 - 1:500. Use reported in scientific literature (PMID 25871979)

In vitro assay

reported in scientific literature (PMID 25411258)

Knockdown Validated

reported in scientific literature (PMID 31655343)

Simple Western

30 ug/ml

Western Blot

2-5 ug/ml

Application Notes

Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min. In human PBMC, a ~120 kDa band is observed. A smaller isoform, TLR9 isoform B (Genbank accession no. AAF72190) containing 975 amino acids may also be observed in some cases. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in Mouse Brain, Ramos lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:50, 30 ug/mL, apparent MW was 90 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 4 reviews rated 3.8 using NBP2-24729 in the following applications:

Immunohistochemistry-Paraffin (1 Review)
Western Blot (3 Reviews)

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Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 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: TLR9

Toll-like receptors (TLRs) are evolutionarily conserved pattern-recognition molecules resembling the toll proteins that mediate antimicrobial responses in Drosophila. These proteins recognize different microbial products during infection and serve as an important link between the innate and adaptive immune responses. TLR9 forms a subfamily along with TLR7 and TLR8 that recognize viral RNA and CpG DNA sequences and are localized in intracellular acidic compartments such as the phagolysosome. Unlike other TLRs which act through adaptor molecules such as TOLLIP, TIRAP, TRIF, and MyD88 to activate various kinases and transcription factors to respond to potential infection, TLR9 is strictly dependent on MyD88.

Long Name

Toll-like Receptor 9

Alternate Names

CD289

Gene Symbol

TLR9

UniProt

Q9NR96

Additional TLR9 Products

Product Documents for TLR9 Antibody (26C593.2) - BSA Free

Download Product Datasheets

Download SDS

Product Specific Notices for TLR9 Antibody (26C593.2) - 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.

Related Research Areas

B Cell Activation

IL-1 Family

NFkB Pathway

NOD-like Receptors and the Inflammasome

Pattern Recognition Receptors

Plasmacytoid Dendritic Cells

Toll-Like Receptors

Citations for TLR9 Antibody (26C593.2) - BSA Free

Customer Reviews for TLR9 Antibody (26C593.2) - BSA Free (4)

3.8 out of 5

4 Customer Ratings

5 Stars

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TLR9 Antibody (26C593.2)

Name: Monica Sauter

Application: Immunohistochemistry-Paraffin

Sample Tested: paraffin embedded macaque retina tissue

Species: macaca mulatta

Verified Customer | Posted 04/12/2018

this antibody recognized TLR9 in paraffin embedded macaque retina tissue

Heat mediated antigen retrieval (pH 6 citrate buffer) 1:25 dilution of primary antibody ON at 4C 1:400 dilution of goat anti-mouse Alexa Fluor 488
TLR9 Antibody (26C593.2)

Name: Anonymous

Application: Western Blot

Sample Tested: human lymphocytes, whole cell lysate

Species: Human

Verified Customer | Posted 10/20/2017

TLR9 in Lympocyte whole cell lysate

3ug/mL in Milk-TBS/T 30ug of protein, 4-12% Bis-Tris gel 10 minute exposure, several bands. 250, 120, 75, 55kDA
Name: Anonymous

Application: Western Blot

Sample Tested: Whole cell lysate

Species: Human

Verified Customer | Posted 01/14/2015
Name: Mariann Kremlitzka

Application: Western Blot

Sample Tested: Primary blood-derived B cells

Species: Human

Verified Customer | Posted 11/25/2014

Expression of surface TLR9 on human B cells

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Showing 1 - 4 的 4 reviews Showing All

Protocols

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

FAQs for TLR9 Antibody (26C593.2) - BSA Free

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Q: I am a bit unsure about your antibody NBP2-24729 specific against TLR9. I used it for probing mouse B cell lysate on Proteisimple WES. On their website the recommend your antibody and claim that it detects a specific signal at 90 kDa. But on your website simple western detects a band at 200 kDa?

A: This came during the race to 1000 and we had many people generating and reviewing data. In retrospect, this was one that probably should not have made it out. As I reviewed the run data, it looks as though the MW ladder did not assign correctly. When this was corrected, the band at 200 actually shifts higher, up to 280 kDa. This band now overlaps with the 280 kDa fluorescent standard which is prone to have non-specific interactions with some antibodies. Those two things make this non-supportive in Simple Western. We ran Raji lysate on the same run. This antibody detects a very broad band with a peak at ~150-165 kDa, closer to expected MW. However, the baseline for this samples was also very high. If we were presented with this data set today, it would not be considered supportive. The other information on the Protein Simple web site likely came from a customer.