Recombinant Anti-Tcl1 antibody [EPR3949] (ab108978)

All lanes : Anti-Tcl1 antibody [EPR3949] (ab108978) at 1/1000 dilution

Lane 1 : Ramos (human Burkitt's lymphoma) whole cell lysate
Lane 2 : Daudi (human Burkitt's lymphoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 13 kDa
Additional bands at: 13 kDa. We are unsure as to the identity of these extra bands.

Blocking buffer: 5% NFDM/TBST

Diluting buffer: 5% NFDM/TBST

ab108978 staining Tcl1 in human tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at 1/500.

Negative control 1: PBS in place of primary antibody.

ab108978 staining Tcl1 in Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab108978 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

All lanes : Anti-Tcl1 antibody [EPR3949] (ab108978) at 1/1000 dilution

Lane 1 : Daudi cell lysate
Lane 2 : Ramos cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Standard HRP labelled goat anti-goat. at 1/2000 dilution

Predicted band size: 13 kDa

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue using ab108978.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ab108978 showing negative staining in Skeletal muscle tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ab108978 showing positive staining in Normal spleen tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ab108978 showing negative staining in Normal brain tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ab108978 showing negative staining in Normal kidney tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ab108978 showing negative staining in Normal stomach tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ab108978 staining Tcl1 in the human cell line Ramos (human Burkitt's lymphoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/400. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

Overlay histogram showing Ramos cells stained with ab108978 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab108978, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Ramos cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.