重组Anti-CD1d抗体[EPR22526-9]

• Flow Cyt

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Flow Cytometry - Anti-CD1d antibody [EPR22526-9] (AB256344)

Flow cytometric analysis of K562 (human chronic myelogenous leukemia lymphoblast, Left) / Jurkat (human T cell leukemia T lymphocyte, Right) cells labeling CD1d with ab256344 at 1/600 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

Negative control : K562 (PMID : 21037579).

• Flow Cyt

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Flow Cytometry - Anti-CD1d antibody [EPR22526-9] (AB256344)

Flow cytometric analysis of human peripheral blood mononuclear cell (PBMC) cells labeling CD1d with ab256344 at 1/600 dilution (0.1μg) (Right) compared with a Rabbit monoclonal IgG (ab172730) (Left) isotype control. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150097) at 1/5000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG (Left) or ab256344 (Right). Then stained with anti-CD19 conjugated to Alexa Fluor^® 647. Gated on viable cells.

• IP

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Immunoprecipitation - Anti-CD1d antibody [EPR22526-9] (AB256344)

CD1d was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate with ab256344 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256344 at 1/1000 dilution (0.6 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

Lane 1 : Jurkat whole cell lysate (10μg)
Lane 2 : ab256344 IP in Jurkat whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab256344 in Jurkat whole cell lysate.

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

Exposure time : 3 mins.

All lanes:

Immunoprecipitation - Anti-CD1d antibody [EPR22526-9] (ab256344)

Predicted band size: 38 kDa

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• WB

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Western blot - Anti-CD1d antibody [EPR22526-9] (AB256344)

The molecular weight observed is consistent with what has been described in the literature (PMID : 10809957).

Negative control : SH-SY5Y (PMID : 26749374), K562 (PMID : 21037579).

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CD1d antibody [EPR22526-9] (ab256344) at 1/1000 dilution

Lane 1:

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

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell 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: 38 kDa

Observed band size: 48 kDa

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Exposure time: 37s