Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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 (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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 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). 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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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 (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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 (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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.

• IP

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Immunoprecipitation - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation.

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) (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 (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] (<a href='/en-us/products/primary-antibodies/mycobacterium-tuberculosis-ag85b-antibody-epr28401-54-ab312328'>ab312328</a>) at 1/1000 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) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation. 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.

• I-ELISA

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Indirect ELISA - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab312328 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

• WB

Supplier Data

Western blot - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] - BSA and Azide free (AB312329)

This data was developed using ab312328, the same antibody clone in a different buffer formulation.

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.

All lanes:

Western blot - Anti-Mycobacterium tuberculosis Ag85B antibody [EPR28401-54] (<a href='/en-us/products/primary-antibodies/mycobacterium-tuberculosis-ag85b-antibody-epr28401-54-ab312328'>ab312328</a>) 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 37 kDa

true

Exposure time: 180s