Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)

Rabbit Recombinant Monoclonal A85B antibody. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra), IP, I-ELISA and reacts with Transfected cell lysate - Human, Transfected cell line, Recombinant fragment samples.

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Images

Western blot - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (AB312328), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	ICC/IF	Flow Cyt (Intra)	IP	I-ELISA
Mycobacterium tuberculosis	Predicted	Predicted	Predicted	Predicted	Predicted	Predicted
Recombinant fragment	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended	Tested
Transfected cell line	Not recommended	Tested	Tested	Tested	Not recommended	Not recommended
Transfected cell lysate - Human	Tested	Not recommended	Not recommended	Not recommended	Tested	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell lysate - Human	Dilution info 1/1000	Notes (0.501ug/ml)

Predicted
Predicted

Species	Dilution info	Notes
Species Mycobacterium tuberculosis	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell line, Recombinant fragment	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell line	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Predicted
Predicted

Species	Dilution info	Notes
Species Mycobacterium tuberculosis	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell lysate - Human, Recombinant fragment	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell line	Dilution info 1/100	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Mycobacterium tuberculosis	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell lysate - Human, Recombinant fragment	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell line	Dilution info 1/500	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Mycobacterium tuberculosis	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell lysate - Human, Recombinant fragment	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Transfected cell lysate - Human	Dilution info 1/30	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Mycobacterium tuberculosis	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell line, Recombinant fragment	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Recombinant fragment	Dilution info 1000 ng/mL	Notes Use at 125 ng/ml

Predicted
Predicted

Species	Dilution info	Notes
Species Mycobacterium tuberculosis	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Transfected cell lysate - Human, Transfected cell line	Dilution info -	Notes -

Associated Products

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1 product for Alternative Version

Target data

See full target information fbpB

Function

The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible for the high affinity of mycobacteria for fibronectin, a large adhesive glycoprotein, which facilitates the attachment of M.tuberculosis to murine alveolar macrophages (AMs). They also help to maintain the integrity of the cell wall by catalyzing the transfer of mycolic acids to cell wall arabinogalactan and through the synthesis of alpha,alpha-trehalose dimycolate (TDM, cord factor). They catalyze the transfer of a mycoloyl residue from one molecule of alpha,alpha-trehalose monomycolate (TMM) to another TMM, leading to the formation of TDM.

Alternative names

Rv1886c, MTCY180.32, fbpB, Diacylglycerol acyltransferase/mycolyltransferase Ag85B, DGAT, 30 kDa extracellular protein, Acyl-CoA:diacylglycerol acyltransferase, Antigen 85 complex B, Extracellular alpha-antigen, Fibronectin-binding protein B, 85B, Ag85B, Fbps B

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR28401-54
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The Mycobacterium tuberculosis Ag85B protein also known as Antigen 85B or FBPA is a component of the Ag85 complex. This protein has a molecular mass of approximately 31 kDa. The Ag85B protein plays an important mechanical role by catalyzing the formation of the cord factor a major glycolipid in the cell wall of Mycobacterium tuberculosis. It is expressed in the mycobacterium's cell wall contributing to the virulence of this pathogen by promoting the invasion and survival within the host.

Biological function summary

The Ag85B protein serves in the assembly and maintenance of the mycobacterial cell wall by synthesizing mycolate. It is a part of the Ag85 complex which includes Ag85A Ag85B and Ag85C ensuring accurate synthesis and export of essential components of the cell wall. Through its actions Ag85B is involved in interacting with the host immune system and is a target for vaccine development due to its immunogenic properties. Its presence is detected during histological examinations of infected tissues where it shows the interaction with host cells.

Pathways

The Ag85B protein functions within the mycolic acid biosynthesis pathway essential for maintaining the structure of the mycobacterial cell envelope. This protein interacts with Ag85A and Ag85C to orchestrate complex lipid synthesis required for Mycobacterium tuberculosis' survival. Ag85B's role in lipid metabolism makes it important in the pathogen's adaptation to the host environment by altering the cell wall to evade immune detection.

Associated diseases and disorders

Ag85B is highly relevant to respiratory diseases like tuberculosis. This protein plays a role in the pathogen's ability to resist destruction by the host's immune system contributing to the chronic nature of tuberculosis. Additionally Ag85B has been investigated in the context of its potential utility in vaccines and diagnostic tests for Mycobacterium tuberculosis infections. Its interactions with other proteins in the Ag85 complex make it a focus of research for developing therapeutics to treat and manage tuberculosis.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Western blot - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This antibody does not cross-react with human fbpA and fbpC.
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328) at 1/1000 dilution
Lane 1: 293T cells transfected with an empty vector containi a His-tag whole cell lysate at 20 µg
Lane 2: 293T cells transfected with a human fbpB expression vector containi a His-tag whole cell lysate at 20 µg
Lane 3: 293T cells transfected with a human fbpA expression vector containi a His-tag whole cell lysate at 20 µg
Lane 4: 293T cells transfected with a human fbpC expression vector containi a His-tag whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 37 kDa
Exposure time: 180s
Flow Cytometry (Intracellular) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells (human embryonic kidney epithelial cell) transfected with a fbpC expression vector containing a myc tag cells labelling Mycobacterium tuberculosis Ag85B with ab312328 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Indirect ELISA - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Indirect ELISA analysis of abab312328 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1:2500 dilution.
Antigen: Mycobacterium tuberculosis Ag85B.
Antigen concentration: 1000 ng/ml
Flow Cytometry (Intracellular) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T cells (human embryonic kidney epithelial cell) transfected with a fbpA expression vector containing a myc tag cells labelling Mycobacterium tuberculosis Ag85B with ab312328 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells (human embryonic kidney epithelial cell) transfected with a fbpB expression vector containing a myc tag (Middle) / 293T cells transfected with an empty expression vector containing a myc tag (Right) cells labelling Mycobacterium tuberculosis Ag85B with ab312328 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Mycobacterium tuberculosis Ag85B with ab312328 at 1/100 (10.01 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing positive staining in 293T cells transfected with a fbpB expression vector containing a myc tag, and no staining in 293T cells transfected with a fbpA or fbpC expression vector containing a myc tag. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Alexa Fluor® 647 Anti-Myc tag antibody [9E10] ab223895 Anti-Myc tag Mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Mycobacterium tuberculosis Ag85B with ab312328 at 1/100 (5.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on mouse liver. The section was incubated with ab312328 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Mycobacterium tuberculosis Ag85B with ab312328 at 1/100 (5.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on rat liver. The section was incubated with ab312328 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Mycobacterium tuberculosis Ag85B with ab312328 at 1/100 (5.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining on human colon. The section was incubated with ab312328 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling Mycobacterium tuberculosis Ag85B with ab312328 at 1/100 (5.01 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T cells transfected with a fbpB expression vector containing a his tag. No staining on (B) HEK-293T cells transfected with a fbpA expression vector containing a his tag, (C) HEK-293T cells transfected with a fbpC expression vector containing a his tag and (D) HEK-293T cells transfected with empty vector containing a his tag. The section was incubated with ab312328 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunoprecipitation - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328)
Mycobacterium tuberculosis Ag85B was immunoprecipitated from 0.35 mg HEK-293T cells (human embryonic kidney epithelial cell) transfected with Human fbpB expression vector containing a his tag whole cell lysate with ab312328 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab312328 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HEK-293T cells (human embryonic kidney epithelial cell) transfected with Human fbpB expression vector containing a his tag whole cell lysate
Lane 2: HEK-293T cells (human embryonic kidney epithelial cell) transfected with Human fbpB expression vector containing a his tag whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab312328 in HEK-293T cells transfected with Human fbpB expression vector containing a his tag whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds
All lanes: Immunoprecipitation - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (ab312328) at 1/30 dilution
All lanes: HEK-293T cells (human embryonic kidney epithelial cell) transfected with Human fbpB expression vector containing a his tag whole cell lysate at 10 µg
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s