Anti-BLBP antibody [EPR24033-13]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling BLBP with ab279649 at 1/500 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum (PMID : 16623952). The section was incubated with ab279649 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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling BLBP with ab279649 at 1/500 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab279649 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Negative control : no staining on human kidney (PMID : 9375786).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunohistochemical analysis of paraffin-embedded human astrocytoma labeling BLBP with ab279649 at 1/500 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human astrocytoma (PMID : 16623952). The section was incubated with ab279649 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) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cells labelling BLBP with ab279649 at 1/250 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in mouse primary gliocyte. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection is observed. Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody was used to counterstain tubulin at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab279649 at a 1/250 dilution followed by ab150120 at a 1/1000 dilution.

Negative control 2 : Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody at a 1/100 dilution followed by ab150077 at a 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling BLBP with ab279649 at 1/2000 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum. The section was incubated with ab279649 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) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum tissue labeling BLBP with ab279649 at 1/50 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebellum, which partially co-stained with GFAP (Red) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1 : 1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized mouse primary neural/glia cells labeling BLBP with ab279649 at 1/500 dilution (Right panel) compared with Rabbit monoclonal IgG Isotype Control (ab172730) (Left panel). Goat anti rabbit IgG (Alexa Fluor^® 647, ab150079) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized rat primary neural/glia cells labeling BLBP with ab279649 at 1/500 dilution (Right panel) compared with Rabbit monoclonal IgG Isotype Control (ab172730) (Left panel). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077 at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling BLBP with ab279649 at 1/2000 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab279649 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) was used.

Heat mediated antigen retrieval 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-BLBP antibody [EPR24033-13] (AB279649)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling BLBP with ab279649 at 1/2000 dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab279649 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Negative control : no staining on mouse kidney.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-BLBP antibody [EPR24033-13] (AB279649)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat primary neural/glia cells labelling BLBP with ab279649 at 1/250 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in rat primary gliocyte. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection is observed. Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody was used to counterstain tubulin at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab279649 at a 1/250 dilution followed by ab150120 at a 1/1000 dilution.

Negative control 2 : Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody at a 1/100 dilution followed by ab150077 at a 1/1000 dilution.

• IP

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Immunoprecipitation - Anti-BLBP antibody [EPR24033-13] (AB279649)

BLBP was immunoprecipitated from 0.35 mg mouse cerebellum tissue lysate 10 μg with ab279649 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279649 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse cerebellum tissue lysate 10 μg

Lane 2 : ab279649 IP in mouse cerebellum tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab279649 in mouse cerebellum tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-BLBP antibody [EPR24033-13] (ab279649)

Predicted band size: 14 kDa

Observed band size: 15 kDa

false

• WB

Lab

Western blot - Anti-BLBP antibody [EPR24033-13] (AB279649)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : spleen, kidney (PMID : 16618771)

Exposure time : 10 seconds

All lanes:

Western blot - Anti-BLBP antibody [EPR24033-13] (ab279649) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human spleen tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

• WB

Lab

Western blot - Anti-BLBP antibody [EPR24033-13] (AB279649)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure times :

Lane 1 : 3 seconds

Lane 2 : 26 seconds

All lanes:

Western blot - Anti-BLBP antibody [EPR24033-13] (ab279649) at 1/1000 dilution

Lane 1:

Rat brain cortex tissue lysate at 20 µg

Lane 2:

Rat cerebellum tissue 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 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

• WB

Lab

Western blot - Anti-BLBP antibody [EPR24033-13] (AB279649)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : spleen, kidney (PMID : 16618771)

Exposure time : 6 seconds

All lanes:

Western blot - Anti-BLBP antibody [EPR24033-13] (ab279649) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse cerebellum tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue 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 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false