Anti-IKZF3 antibody [EPR9342(B)]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

Intracellular Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma cell line cells labeling IKZF3 with purified ab139408 at 1/90 (red).

Cells were fixed with 2% paraformaldehyde. An FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

Immunocytochemistry/Immunofluorescence analysis of Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling IKZF3 (green) with purified ab139408 at 1/100.

Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control : Secondary antibody ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labeling IKZF3 with unpurified ab139408 at 1/250 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labeling IKZF3 with purified ab139408 at 1/100.

Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue labeling IKZF3 with purified ab139408 at 1/100.

Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue labeling IKZF3 with purified ab139408 at 1/100.

Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Unknown

Western blot - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

All lanes:

Western blot - Anti-IKZF3 antibody [EPR9342(B)] (ab139408) at 1/1000 dilution

Lane 1:

Human thymus cell lysate at 10 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 10 µg

Lane 3:

Raji (Human Burkitt's lymphoma cell line) cell lysate at 10 µg

Lane 4:

Ramos (Human Burkitt's lymphoma cell line) cell lysate at 10 µg

Secondary

All lanes:

HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 58 kDa

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• WB

Lab

Western blot - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IKZF3 antibody [EPR9342(B)] (ab139408) at 1/20000 dilution

Lane 1:

Raji (Human Burkitt's lymphoma cell line) cell lysate at 20 µg

Lane 2:

Ramos (Human Burkitt's lymphoma cell line) cell lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg

Secondary

All lanes:

Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 58 kDa

Observed band size: 70 kDa

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• WB

CiteAb

Western blot - Anti-IKZF3 antibody [EPR9342(B)] (AB139408)

IKZF3 western blot using anti-IKZF3 antibody [EPR9342(B)] ab139408. Publication image and figure legend from Mori, T., Ito, T., et al., 2018, Sci Rep, PubMed 29358579.

ab139408 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab139408 please see the product overview.

Effects of deuterium-substituted (S)- and (R)-thalidomides on IKZF3 degradation and CRBN-IKZF3 binding. (a) Effects of deuterated (S)- and (R)-thalidomide enantiomers, (S)-D-Thal and (R)-D-Thal, on IKZF3 protein levels in MM1S cells after 12-h treatment. IB, immunoblot. (b) Co-immunoprecipitation of FLAG-HA-CRBN in the presence of each deuterated thalidomide enantiomer, (S)-D-Thal or (S)-D-Thal. IP, immunoprecipitation. Full-length blots in (a) and (b) are presented in Supplementary Fig. 9.

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