Anti-CD3 zeta antibody [BL-336-1B2]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

Immunohistochemical analysis of Formalin-Fixed Paraffin-Embedded human appendix tissue labeling human CD247/CD3Z with ab243874, followed by a HRP-conjugated goat anti-rabbit IgG as secondary. Substrate : DAB. Membrane staining on human appendix is observed. Counter stained with hematoxylin. Epitope retrieval with citrate buffer pH 6.0 for 20 minutes is recommended.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue, labeling CD247/CD3Z with ab243874. HRP-conjugated goat anti-rabbit IgG used as the secondary antibody. DAPI counterstain.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human lung cancer tissue labeling CD247/CD3Z with ab243874. HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody, with DAB as the substrate.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

Immunohistochemical analysis of Formalin-Fixed Paraffin-Embedded human tonsil tissue labeling human CD247/CD3Z with ab243874, followed by a HRP-conjugated goat anti-rabbit IgG as secondary. Substrate : DAB. Membrane staining on human tonsil is observed. Counter stained with hematoxylin. Epitope retrieval with citrate buffer pH 6.0 for 20 minutes is recommended.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD247/CD3Z with ab243874. HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody, with DAB as substrate.

• IP

Supplier Data

Immunoprecipitation - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

CD247/CD3Z was immunoprecipitated from 1mg of Jurkat whole cell lysate with ab243874 at 20 μl/mg lysate. Western blot was performed from the immunoprecipitate using ab264158 at a 1/1000 dilution.

Lane 1 : ab243874 IP in Jurkat whole cell lysate.
Lane 2 : Control IgG.

Detection : Chemiluminescence.

Lysate prepared using NETN lysis buffer.

All lanes:

Immunoprecipitation - Anti-CD3 zeta antibody [BL-336-1B2] (ab243874)

Predicted band size: 19 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

False colour image of Western blot : Anti-CD3 zeta antibody [BL-336-1B2] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab243874 was shown to bind specifically to CD3 zeta. A band was observed at 16 kDa in wild-type Jurkat cell lysates with no signal observed at this size in CD247 knockout cell line ab273856 (knockout cell lysate ab273810). To generate this image, wild-type and CD247 knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-CD3 zeta antibody [BL-336-1B2] (ab243874) at 1/1000 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

CD247 knockout Jurkat cell lysate at 20 µg

Lane 2:

Western blot - Human CD247 knockout Jurkat cell line (<a href='/ko/products/cell-lines/human-cd247-knockout-jurkat-cell-line-ab273856'>ab273856</a>)

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 19 kDa

Observed band size: 16 kDa

false

• WB

Supplier Data

Western blot - Anti-CD3 zeta antibody [BL-336-1B2] (AB243874)

Lysates prepared using NETN lysis buffer.

All lanes:

Western blot - Anti-CD3 zeta antibody [BL-336-1B2] (ab243874) at 1/1000 dilution

Lane 1:

K562 whole cell lysate at 50 µg

Lane 2:

Jurkat whole cell lysate at 50 µg

Lane 3:

HeLa whole cell lysate at 50 µg

Lane 4:

SR whole cell lysate at 50 µg

Lane 5:

HEK293T whole cell lysate at 50 µg

Lane 6:

HepG2 whole cell lysate at 50 µg

Lane 7:

MOLT4 whole cell lysate at 50 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG

Predicted band size: 19 kDa

false

Exposure time: 10s