Anti-CD40 Ligand antibody [EPR26530-17]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (AB303610)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type Jurkat tissue labeling CD40 Ligand with ab303610 at 1/500 (1.062 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Positive staining on wild-type Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cell pellet (A) and Jurkat treated with 50ng/ml 48h PMA for 48h, then treated with 300ng/ml BFA and 3¦МM monensin for 24h cell pellet (B). No staining on wild-type Raji (human Burkitt's lymphoma B lymphocyte) cell pellet. The section was incubated with ab303610 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (AB303610)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD40 Ligand with ab303610 at 1/500 (1.062 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection) was used. Positive staining on human tonsil (PMID : 7699321). The section was incubated with ab303610 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-CD40 Ligand antibody [EPR26530-17] (AB303610)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling CD40 Ligand with ab303610 at 1/500 (1.062 ug/ml) followed by a ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Negative control : No staining on human skeletal muscle (PMID : 1281209). The section was incubated with ab303610 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

• IP

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Immunoprecipitation - Anti-CD40 Ligand antibody [EPR26530-17] (AB303610)

CD40 Ligand was immunoprecipitated from 0.35 mg D1.1 (human T lymphoblast) whole cell lysate 10 ug with ab303610 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab303610 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : D1.1 (human T lymphoblast) whole cell lysate 10 ug
Lane 2 : ab303610 IP in D1.1 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab303610 in D1.1 whole cell lysate

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

Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610) at 1/1000 dilution

Lane 1:

D1.1 (human T lymphoblast) whole cell lysate 10 μg

Lane 2:

D1.1 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 17 kDa,25 kDa,30 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (AB303610)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : human skeletal muscle(PMID : 1281209) The antibody recognizes both membrane bound(25-30KDa) and soluble(17KDa) form. Exposure time : 180 seconds

All lanes:

Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610) at 1/1000 dilution

Lane 1:

Human lymphma tissue lysate 50 μg

Lane 2:

Human spleen tissue lysate 50 μg

Lane 3:

Human tonsil tissue lysate 50 μg

Lane 4:

Human skeletal muscle tissue lysate 50 μg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Observed band size: 17 kDa,25 kDa,30 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (AB303610)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : Raji(PMID : 12805559) The antibody recognizes both membrane bound(25-30KDa) and soluble(17KDa) form. Exposure time : 3.25 seconds

All lanes:

Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610) at 1/1000 dilution

Lane 1:

Untreated Jurkat (human T cell leukemia T lymphocyte from peripheral blood), whole cell lysate 50 μg

Lane 2:

Jurkat treated with /ml 48h PMA for 48h, then treated with 300ng/ml BFA and 3uM monensin for 24h, whole cell lysate 50 μg

Lane 3:

D1.1 (human T lymphoblast), whole cell lysate 50 μg

Lane 4:

Raji (human Burkitts lymphoma B lymphocyte), whole cell lysate 50 μg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Observed band size: 17 kDa,25 kDa,30 kDa

false

Exposure time: 3.25s