Anti-CD1a antibody [EPR26526-62]

• Flow Cyt

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Flow Cytometry - Anti-CD1a antibody [EPR26526-62] (AB313875)

Flow cytometric analysis of HeLa (human cervical adenocarcinoma epithelial cell, Left) / MOLT-4 (human lymphoblastic leukemia T lymphoblast, Right) cells labelling CD1a with ab313875 at 1/50 dilution (1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : HeLa. Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD1a antibody [EPR26526-62] (AB313875)

Immunohistochemical analysis of paraffin-embedded human thymus tissue labeling CD1a with ab313875 at 1/10000 (0.052 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on cortical thymocytes of human thymus (PMID : 28400115; PMID : 21565561). (A) Low‑powered (magnification, x100) and (B) high‑powered (magnification, x400) microscopic images. The section was incubated with ab313875 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). 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-CD1a antibody [EPR26526-62] (AB313875)

Immunohistochemical analysis of paraffin-embedded human thymoma tissue labeling CD1a with ab313875 at 1/10000 (0.052 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human thymoma (PMID : 35547871). The section was incubated with ab313875 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). 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-CD1a antibody [EPR26526-62] (AB313875)

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling CD1a with ab313875 at 1/10000 (0.052 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on epidermal Langerhans cells of human skin (PMID : 20514853). The section was incubated with ab313875 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). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

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Multiplex immunohistochemistry - Anti-CD1a antibody [EPR26526-62] (AB313875)

Fluorescence multiplex immunohistochemical analysis of the human skin (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-CD1A (green; Opal™520) and anti-CD207 (red; Opal™570) on human skin. Panel B : anti-CD1A stained on Langerhans cells subsets. Panel C : anti-CD207 stained on Langerhans cells subsets. The section was incubated in two rounds of staining : in the order of ab283686 and ab313875 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt

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Flow Cytometry - Anti-CD1a antibody [EPR26526-62] (AB313875)

Flow cytometric analysis of CD1A KO Jurkat (CD1A knockout human T cell leukemia T lymphocyte from peripheral blood, Left) / Parental Jurkat (Right) cells labelling CD1a with ab313875 at 1/50 dilution (1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD1a antibody [EPR26526-62] (AB313875)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling CD1a with ab313875 at 1/10000 (0.052 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human cerebrum (PMID : 27666391). The section was incubated with ab313875 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). 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-CD1a antibody [EPR26526-62] (AB313875)

CD1a was immunoprecipitated from 0.35 mg Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate with ab313875 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313875 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate Lane 2 : ab313875 IP in Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313875 in Jurkat whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-CD1a antibody [EPR26526-62] (ab313875) at 1/30 dilution

All lanes:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 15s

• WB

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Western blot - Anti-CD1a antibody [EPR26526-62] (AB313875)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : HeLa. In Western blot, ab313875 was shown to bind specifically to CD1A. Target of interest was observed at 45-50 kDa in wild-type Jurkat cell lysates (lane 1) with no signal observed at this size in CD1A knockout cell line (lane 2) (lane 2, knockout cell line ab274926). Samples are non-boiled as boiling may cause protein aggregation. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD1a antibody [EPR26526-62] (ab313875) at 1/1000 dilution

Lane 1:

Wild-type Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 60 µg

Lane 2:

CD1A knockout Jurkat whole cell lysate at 60 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 60 µg

Lane 4:

MOLT-4 (human lymphoblastic leukemia t lymphoblast) whole cell lysate at 60 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 45-50 kDa

false

Exposure time: 8s