TLR9 antibody (AA 868-1016)
(2 validations)-
- Target See all TLR9 Antibodies
- TLR9 (Toll-Like Receptor 9 (TLR9))
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Binding Specificity
- AA 868-1016
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Reactivity
- Human
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Host
- Mouse
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Clonality
- Monoclonal
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Conjugate
- This TLR9 antibody is un-conjugated
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Application
- Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Flow Cytometry (FACS), Immunocytochemistry (ICC)
- Purification
- purified
- Immunogen
- Purified recombinant fragment of human TLR9 (AA: 868-1016) expressed in E. coli.
- Clone
- 1B12H2
- Isotype
- IgG2a
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anti-Toll-Like Receptor 9 (TLR9) (AA 28-362) antibody
TLR9 Reactivity: Human WB, IHC, ICC, IP Host: Rabbit Polyclonal unconjugated
anti-Toll-Like Receptor 9 (TLR9) (AA 26-193) antibodyTLR9 Reactivity: Human WB, IHC, ELISA, FACS, ICC, Neut Host: Mouse Monoclonal 5B10E8 unconjugated
anti-Toll-Like Receptor 9 (TLR9) (AA 350-450) antibodyTLR9 Reactivity: Human WB, IHC Host: Rabbit Polyclonal unconjugated
anti-Toll-Like Receptor 9 (TLR9) (AA 268-300) antibodyTLR9 Reactivity: Human, Mouse, Rat, Monkey WB, IHC, ELISA, FACS, IHC (p), ICC, IP, IF Host: Mouse Monoclonal 26C593-2 unconjugated
anti-Toll-Like Receptor 9 (TLR9) (AA 951-1032) antibodyTLR9 Reactivity: Human, Mouse, Rat WB, ELISA, FACS, IHC (p), IHC (fro), IF (p), IF (cc) Host: Rabbit Polyclonal unconjugated
anti-Toll-Like Receptor 9 (TLR9) antibody (Biotin)TLR9 Reactivity: Human WB, FACS, IHC (p), IF Host: Mouse Monoclonal 5G5 Biotin
anti-Toll-Like Receptor 9 (TLR9) (N-Term) antibodyTLR9 Reactivity: Human WB, IHC (p) Host: Rabbit Polyclonal unconjugated
anti-Toll-Like Receptor 9 (TLR9) (AA 99-215) antibodyTLR9 Reactivity: Human WB, ELISA, IF Host: Mouse Monoclonal 1E8 unconjugated
anti-Toll-Like Receptor 9 (TLR9) antibodyTLR9 Reactivity: Human WB, FACS, IHC (p), IF Host: Mouse Monoclonal 5G5 unconjugated
anti-Toll-Like Receptor 9 (TLR9) antibody (FITC)TLR9 Reactivity: Human WB, FACS, IHC (p), IF Host: Mouse Monoclonal 5G5 FITC
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- Application Notes
- ELISA: 1:10000, WB: 1:500 - 1:2000, IHC: 1:200 - 1:1000, ICC: 1:200 - 1:1000, FCM: 1:200 - 1:400
- Restrictions
- For Research Use only
-
- by
- Klinik für Anästhesie, Intensivmedizin und Schmerztherapie, Universitätsklinikum der Ruhr Universität Bochum
- No.
- #103693
- Date
- 03/26/2019
- Antigen
- TLR9
- Lot Number
- 160127
- Method validated
- Immunofluorescence
- Positive Control
U937 monocytic cells
- Negative Control
No primary antibody control
- Notes
Passed. ABIN5542492 localization on ER is visible. Negative control without primary antibody shows no signal.
- Primary Antibody
- ABIN5542492
- Secondary Antibody
- AF488-conjugated AffiniPure alpaca anti-Mouse IgG antibody (Jackson ImmunoResearch, 615-545-214)
- Full Protocol
- Grow U937 cells in RPMI (Gibco, 21875-034) supplemented with 10% FCS (Gibco, A3840001) and Pen/Strep(Gibco, 15140-122), at 37°C and 5% CO2 in 500µL in 12-well plates.
- Use cytospin to spin cells onto slide.
- Fix cells on coverslide in 4% PFA for 20min at RT.
- Wash cells 3x for 5min with PBS.
- Permeabilize cells in PBS containing 0.1% Triton for 5min at RT.
- Block non-specific binding with Duolink PLA Blocking Solution (Sigma) for 30 min at RT.
- Incubate cells with primary mouse anti-TLR9 antibody (antibodies-online, ABIN5542492, lot 160127) diluted 1:200 in staining solution ON at 4°C.
- Wash cells 3x for 5min with PBS.
- Incubate cells with secondary AF488-conjugated AffiniPure alpaca anti-Mouse IgG antibody (Jackson ImmunoResearch, 615-545-214) diluted 1:400 in staining solution for 1h at RT.
- Wash cells 2x for 5min with PBS.
- Mount coverslips on glass slides in ProLong Gold antifade reagent (Invitrogen, P36935,1890418) containing DAPI.
- Image acquisition with an Olympus widefield microscope.
- Experimental Notes
TLR9 staining using ABIN5542492 shows typical localization peri-nuclear localization.
Validation #103693 (Immunofluorescence)!['Independent Validation' Badge]( "Successfully validated")
!['Independent Validation' Badge]( "Successfully validated")
Validation Images![U937 monocytic cells stained with with ABIN5542492 (top row). TLR9 staining (AF488, green) shows typical peri-nuclear localization (ER). Counterstain with DAPI (blue). Negative control without primary antibody (bottom row) shows no significant staining. Counterstain with DAPI (blue).]()
U937 monocytic cells stained with with ABIN5542492 (top row). TLR9 staining (AF488, green) shows typical peri-nuclear localization (ER). Counterstain with DAPI (blue). Negative control without primary antibody (bottom row) shows no significant staining. Counterstain with DAPI (blue).
![Negative controls of U937 cells without either ABIN5542492 or ABIN361758 show no signal. In U937 cells incubated simulatnously with ABIN5542492 and ABIN361758 the PLA signal (green) is clearly visible. Counterstain with DAPI (blue).]()
Negative controls of U937 cells without either ABIN5542492 or ABIN361758 show no signal. In U937 cells incubated simulatnously with ABIN5542492 and ABIN361758 the PLA signal (green) is clearly visible. Counterstain with DAPI (blue).
Full Methods -
- by
- Klinik für Anästhesie, Intensivmedizin und Schmerztherapie, Universitätsklinikum der Ruhr Universität Bochum
- No.
- #103821
- Date
- 04/29/2019
- Antigen
- TLR9
- Lot Number
- 160127
- Method validated
- Proximity Ligation Assay
- Positive Control
U937 monocytic cells stimulated with ODN1826
- Negative Control
Anti TLR9 antibody (ABIN5542492) alone
Anti pY antibody (ABIN361758) alone
- Notes
Passed. PLA signal visible in stimulated cells. Negative control with only one primary antibody shows no signal.
- Primary Antibody
- ABIN5542492
- Secondary Antibody
- ABIN361758
- Full Protocol
- Grow U937 cells in RPMI (Gibco, 21875-034) supplemented with 10% FCS (Gibco, A3840001) and Pen/Strep(Gibco, 15140-122), at 37°C and 5% CO2 in 500µL in 12-well plates.
- Incubate the cells with 1µM ODN1826 for 4h.
- Use cytospin to spin cells onto slide.
- Fix cells on coverslide in 4% PFA for 20min at RT.
- Wash cells 3x for 5min with PBS.
- Permeabilize cells in PBS containing 0.1% Triton for 5min at RT.
- Block non-specific binding with Duolink PLA Blocking Solution (Sigma-Aldrich) for 30min at RT.
- Incubate cells with primary mouse anti-TLR9 antibody (antibodies-online, ABIN5542492, lot 160127) diluted 1:200 and primary rabbit anti-pY antibody (antibodies-online, ABIN361758, lot 1508290) diluted 1:100 ON at 4°C.
- Wash cells 3x for 5min with PBS.
- Incubate cells with Duolink In Situ PLA Probe anti-Rabbit MINUS (Sigma-Aldrich, DUO92005, lot SLBZ4516) and Duolink In Situ PLA Probe anti-Mouse mouse PLUS (Sigma-Aldrich, DUO92001, lot SLBZ8369) proximity probes according to manufacturers recommendations.
- Wash cells 2x for 5min with PBS.
- Perform Ligation step according to the Duolink PLA kit (Sigma).
- Perform Amplification step according to the Duolink In Situ PlA Detection Kit Green (Sigma-Aldrich, DUO92014).
- Mount coverslips on glass slides in ProLong Gold antifade reagent (Invitrogen, P36935, lot 1890418) containing DAPI.
- Image acquisition with an Olympus widefield microscope.
- Experimental Notes
We used double the amount the Phi29 polymerase for the amplifaction step than recommended.
Validation #103821 (Proximity Ligation Assay)!['Independent Validation' Badge]( "Successfully validated")
!['Independent Validation' Badge]( "Successfully validated")
Validation Images![U937 monocytic cells stained with with ABIN5542492 (top row). TLR9 staining (AF488, green) shows typical peri-nuclear localization (ER). Counterstain with DAPI (blue). Negative control without primary antibody (bottom row) shows no significant staining. Counterstain with DAPI (blue).]()
U937 monocytic cells stained with with ABIN5542492 (top row). TLR9 staining (AF488, green) shows typical peri-nuclear localization (ER). Counterstain with DAPI (blue). Negative control without primary antibody (bottom row) shows no significant staining. Counterstain with DAPI (blue).
![Negative controls of U937 cells without either ABIN5542492 or ABIN361758 show no signal. In U937 cells incubated simulatnously with ABIN5542492 and ABIN361758 the PLA signal (green) is clearly visible. Counterstain with DAPI (blue).]()
Negative controls of U937 cells without either ABIN5542492 or ABIN361758 show no signal. In U937 cells incubated simulatnously with ABIN5542492 and ABIN361758 the PLA signal (green) is clearly visible. Counterstain with DAPI (blue).
Full Methods -
- Format
- Liquid
- Buffer
- Purified antibody in PBS with 0.05 % sodium azide
- Preservative
- Sodium azide
- Precaution of Use
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Storage
- 4 °C/-20 °C
- Storage Comment
- 4°C, -20°C for long term storage
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- Target
- TLR9 (Toll-Like Receptor 9 (TLR9))
- Alternative Name
- TLR9 (TLR9 Products)
- Synonyms
- CD289 antibody, toll like receptor 9 antibody, toll-like receptor 9 antibody, TLR9 antibody, Tlr9 antibody
- Background
-
Description: The protein encoded by this gene is a member of the Toll-like receptor (TLR) family which plays a fundamental role in pathogen recognition and activation of innate immunity. TLRs are highly conserved from Drosophila to humans and share structural and functional similarities. They recognize pathogen-associated molecular patterns (PAMPs) that are expressed on infectious agents, and mediate the production of cytokines necessary for the development of effective immunity. The various TLRs exhibit different patterns of expression. This gene is preferentially expressed in immune cell rich tissues, such as spleen, lymph node, bone marrow and peripheral blood leukocytes. Studies in mice and human indicate that this receptor mediates cellular response to unmethylated CpG dinucleotides in bacterial DNA to mount an innate immune response.,
Aliases: CD289
- Molecular Weight
- 115.8 kDa
- Gene ID
- 54106
- HGNC
- 54106
- Pathways
- TLR Signaling, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Toll-Like Receptors Cascades
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