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Antibodies

Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) Recombinant Rabbit monoclonal Antibody

Product Sizes
100μl
£425.00
HA722499-100UL
About this Product
SKU:
HA722499
Application:
Western Blot, Flow Cytometry
Buffer:
PBS (pH7.4); 0.1% BSA; 40% Glycerol. Preservative: 0.05% Sodium Azide.
translate.label.attr.clone:
PSH06-00
Clonality:
Monoclonal
Conjugate:
Non-conjugated
Host:
Rabbit
Isotype:
IgG
Physical State:
Liquid
Purity:
Protein A affinity purified.
Shipping Conditions:
Blue Ice
Storage Conditions:
Shipped at 4[o]C. Store at +4[o]C short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20[o]C long term.
Supplier:
Huabio
Type:
Antibodies: Monoclonal Antibody
Western blot analysis of Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) on different lysates with Rabbit anti-Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) antibody (HA722499) at 1/1;000 dilution. Lane 1: Jurkat cell lysate Lane 2: Jurkat treated with 20μM LY294002 for 2 hours cell lysate Lane 3: Jurkat treated with 100nM Calyculin A for 40 minutes cell lysate Lane 4: NIH/3T3 starved for 24 hours cell lysate Lane 5: NIH/3T3 starved for 24 hours then add 100nM Calyculin A for 30 minutes cell lysate Lysates/proteins at 20 µg/Lane. Observed band size: 55-150 kDa Exposure time: 20 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722499) at 1/1;000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50;000 dilution was used for 1 hour at room temperature.
Flow cytometric analysis of Jurkat cells treated with 20μM LY294002 for 2 hours (left; negative) and Jurkat cells treated with 100nM Calyculin A for 40 minutes (right; positive) labeling Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26). Cells were fixed and permeabilized. Then stained with the primary antibody (HA722499; 1/1;000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour; the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1;000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Immunocytochemistry analysis of Jurkat cells treated with 20μM LY294002 for 2 hours (upper; negative) and Jurkat cells treated with 100nM Calyculin A for 40 minutes (lower; positive) labeling Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) with Rabbit anti-Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) antibody (HA722499) at 1/1;000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature; permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature; then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-FOXO1A (T24)+ FOXO3A (T32)+ FOXO4 (T28)+ FOXO6 (T26) antibody (HA722499) at 1/1;000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488; HA1121) was used as the secondary antibody at 1/1;000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2; red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594; HA1126) was used as the secondary antibody at 1/1;000 dilution.