Anti-EBI3 antibody [EPR25341-155] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EBI3 antibody [EPR25341-155] - BSA and Azide free (AB307196)

This data was developed using ab307195, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling EBI3 with ab307195 at 1/2000 (0.26 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used. Positive staining on human placenta (PMID : 11696437). The section was incubated with ab307195 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 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-EBI3 antibody [EPR25341-155] - BSA and Azide free (AB307196)

This data was developed using ab307195, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling EBI3 with ab307195 at 1/2000 (0.26 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used. Negative control : No staining on human cardiac muscle. The section was incubated with ab307195 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EBI3 antibody [EPR25341-155] - BSA and Azide free (AB307196)

This data was developed using ab307195, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized HDLM-2 (human Hodgkin lymphoma cell) cells labelling EBI3 with ab307195 at 1/50 (10.38 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 cytoplasmic staining in HDLM-2 cell line. Negative control : Jurkat (PMID : 15793300). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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.

• WB

Supplier Data

Western blot - Anti-EBI3 antibody [EPR25341-155] - BSA and Azide free (AB307196)

This data was developed using 307195, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Negative control : human heart (PMID : 8551575). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 8551575). The identity of the higher MW band at approximately 50 kDa (in lane 1) is unknown. 26 seconds Exposure time :

All lanes:

Western blot - Anti-EBI3 antibody [EPR25341-155] (<a href='/en-us/products/primary-antibodies/ebi3-antibody-epr25341-155-ab307195'>ab307195</a>) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate 20 μg

Lane 2:

Human heart tissue lysate 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: 27 kDa,34 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-EBI3 antibody [EPR25341-155] - BSA and Azide free (AB307196)

This data was developed using 307195, the same antibody clone in a different buffer formulation.

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

Negative control : Jurkat (PMID : 11696437), MOLT-4 (PMID : 11696437).

EBI3 is a glycoprotein of approximately 34 kDa and detected as a 27 kDa band after treated with Protein Deglycosylation MIX II.

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-EBI3 antibody [EPR25341-155] (<a href='/en-us/products/primary-antibodies/ebi3-antibody-epr25341-155-ab307195'>ab307195</a>) at 1/1000 dilution

Lane 1:

Untreated HDLM-2 (human Hodgkin lymphoma) whole cell lysate 20 μg

Lane 2:

HDLM-2 whole cell lysate treated with Protein Deglycosylation Mix II 20 μg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 20 μg

Lane 4:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate 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: 27 kDa,34 kDa

true

Exposure time: 180s