Anti-CD1a antibody [EP3622]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD1a antibody [EP3622] (AB108309)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymoma tissue sections labelling CD1a with purified ab108309 at 1/1000 dilution (0.14 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Lab

Western blot - Anti-CD1a antibody [EP3622] (AB108309)

The molecular weight observed is consistent with what has been described in the literature (PMID : 18178838).

All lanes:

Western blot - Anti-CD1a antibody [EP3622] (ab108309) at 1/1000 dilution

All lanes:

Human thymus lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 37 kDa

Observed band size: 49 kDa

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• WB

Lab

Western blot - Anti-CD1a antibody [EP3622] (AB108309)

False colour image of Western blot : Anti-CD1a antibody [EP3622] 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, ab108309 was shown to bind specifically to CD1a. A band was observed at 45-50 kDa in wild-type Jurkat cell lysates with no signal observed at this size in CD1A knockout cell line ab274926 (knockout cell lysate ab274984). To generate this image, wild-type and CD1A 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-CD1a antibody [EP3622] (ab108309) at 1/1000 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

CD1A knockout Jurkat cell lysate at 20 µg

Lane 2:

Western blot - Human CD1A knockout Jurkat cell line (<a href='/en-us/products/cell-lines/human-cd1a-knockout-jurkat-cell-line-ab274926'>ab274926</a>)

Lane 3:

MOLT-4 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 37 kDa

Observed band size: 45-50 kDa

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