Anti-Tcl1 antibody [EPR3949]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Tcl1 antibody [EPR3949] (AB108978)

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 >5000 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.

• WB

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Western blot - Anti-Tcl1 antibody [EPR3949] (AB108978)

All lanes:

Western blot - Anti-Tcl1 antibody [EPR3949] (ab108978) at 1/1000 dilution

Lane 1:

Daudi cell lysate at 10 µg

Lane 2:

Ramos cell lysates at 10 µg

Secondary

All lanes:

Standard HRP labelled goat anti-goat. at 1/2000 dilution

Predicted band size: 13 kDa

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• WB

Lab

Western blot - Anti-Tcl1 antibody [EPR3949] (AB108978)

Blocking buffer : 5% NFDM/TBST

Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Tcl1 antibody [EPR3949] (ab108978) at 1/1000 dilution

Lane 1:

Ramos (human Burkitt's lymphoma) whole cell lysate at 20 µg

Lane 2:

Daudi (human Burkitt's lymphoma) whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 103 kDa,13 kDa,43 kDa

Observed band size: 13 kDa,27 kDa,43 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tcl1 antibody [EPR3949] (AB108978)

ab108978 showing negative staining in Normal kidney tissue.

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Tcl1 antibody [EPR3949] (AB108978)

ab108978 staining Tcl1 in the human cell line Ramos (human Burkitt's lymphoma) by intracellular 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)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tcl1 antibody [EPR3949] (AB108978)

ab108978 showing negative staining in Skeletal muscle tissue. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tcl1 antibody [EPR3949] (AB108978)

ab108978 showing negative staining in Normal stomach tissue.

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tcl1 antibody [EPR3949] (AB108978)

ab108978 showing positive staining in Normal spleen tissue.

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Tcl1 antibody [EPR3949] (AB108978)

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)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tcl1 antibody [EPR3949] (AB108978)

ab108978 showing negative staining in Normal brain tissue.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tcl1 antibody [EPR3949] (AB108978)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tcl1 antibody [EPR3949] (AB108978)

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

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