重组Anti-CD134 / OX40L receptor抗体[EPR22229-5]

• Flow Cyt

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Flow Cytometry - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (AB229021)

Flow cytometric analysis of rat primary splenocytes treated with 2.5μg/ml concanavalin A for 72h labeling CD134 / OX40L with ab229021 at 1/500 dilution (Right) compared to cells stained with Alexa Fluor^® 647-conjugated CD3 and rabbit IgG (Left). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at 1/2000 dilution.

Data shown were gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (AB229021)

Flow cytometric analysis of mouse primary splenocytes treated with 2.5μg/ml concanavalin A for 72h labeling CD134 / OX40L with ab229021 at 1/500 dilution (Right) compared to cells stained with Alexa Fluor® 647-conjugated CD3 and rabbit IgG (Left). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at 1/2000 dilution.

Data shown were gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (AB229021)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD134 / OX40L using ab229021 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on T cells in rat spleen (PMID : 15259024; PMID : 10233901). Counterstained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins.

The section was incubated with ab229021 for 10 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (AB229021)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD134 / OX40L using ab229021 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on T cells in periarterial lymphatic sheath of mouse spleen (PMID : 15259024; PMID : 10233901). Counterstained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins.

The section was incubated with ab229021 for 10 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

• IP

Unknown

Immunoprecipitation - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (AB229021)

CD134 / OX40L was immunoprecipitated from 0.35 mg rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate using ab229021 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab229021 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at a 1/5000 dilution.

Lane 1 : Rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate 10 μg (input)
Lane 2 : ab229021 IP in Rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab229021 in Rat splenocytes treated with 2.5μg/ml concanavalin A for 72h lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 20 seconds.

All lanes:

Immunoprecipitation - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (ab229021)

Predicted band size: 29 kDa

false

• WB

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Western blot - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (AB229021)

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

CD134 / OX40L is a glycosylated protein and its expression is increased after concanavalin A treatment. PMID 23897980; PMID 17609274

All lanes:

Western blot - Anti-CD134 / OX40L receptor antibody [EPR22229-5] (ab229021) at 1/1000 dilution

Lane 1:

Untreated mouse splenocytes lysate at 20 µg

Lane 2:

Mouse splenocytes treated with 2.5ug/ml concanavalin A for 72h lysate at 20 µg

Lane 3:

Untreated rat splenocytes lysate at 20 µg

Lane 4:

Rat splenocytes treated with 2.5ug/ml concanavalin A for 72h lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 29 kDa

Observed band size: 50 kDa

false

Exposure time: 3min