Aly Recombinant Rabbit Monoclonal Antibody [JE36-45]
cat.: HA721961
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE36-45
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 27 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Aly aa 208-257 / 257.
Positive control: HepG2 cell lysate, K-562 cell lysate, 293T cell lysate, A549 cell lysate, SK-OV-3 cell lysate, HL-60 cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, C6 cell lysate, mouse testis tissue lysate, mouse spleen tissue lysate, rat testis tissue lysate, HepG2, human brain tissue, mouse brain tissue, rat brain tissue.
Subcellular location: Nucleus, Nucleus speckle, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:1,000
1:250
1:2,000
1:1,000
Uniprot #: SwissProt: Q86V81 Human | O08583 Mouse
Entrez Gene: 690585 Rat
Alternative names: Ally of AML-1 and LEF-1 Ally of AML1 and LEF1 ALY ALY/REF Aly/REF export factor BEF bZIP enhancing factor bZIP-enhancing factor BEF REF THO complex 4 THO complex subunit 4 Tho4 thoc4 THOC4_HUMAN Transcriptional coactivator Aly/REF Transcriptional coactivator
Images
HA721961_1.jpg Fig1: Western blot analysis of Aly on different lysates with Rabbit anti-Aly antibody (HA721961) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: K-562 cell lysate (20 µg/Lane)
Lane 3: 293T cell lysate (20 µg/Lane)
Lane 4: A549 cell lysate (20 µg/Lane)
Lane 5: SK-OV-3 cell lysate (20 µg/Lane)
Lane 6: HL-60 cell lysate (20 µg/Lane)
Lane 7: NIH/3T3 cell lysate (20 µg/Lane)
Lane 8: Neuro-2a cell lysate (20 µg/Lane)
Lane 9: C6 cell lysate (20 µg/Lane)
Lane 10: Mouse testis tissue lysate (40 µg/Lane)
Lane 11: Mouse spleen tissue lysate (40 µg/Lane)
Lane 12: Rat testis tissue lysate (40 µg/Lane)

Predicted band size: 27 kDa
Observed band size: 27 kDa

Exposure time: 2 minutes;

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 (HA721961) 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.
HA721961_2.jpg Fig2: Immunocytochemistry analysis of HepG2 cells labeling Aly with Rabbit anti-Aly antibody (HA721961) at 1/250 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-Aly antibody (HA721961) at 1/250 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.
HA721961_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-Aly antibody (HA721961) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721961) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721961_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Aly antibody (HA721961) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721961) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721961_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Aly antibody (HA721961) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721961) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721961_6.jpg Fig6: Flow cytometric analysis of HepG2 cells labeling Aly.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721961, 1μg/mL) (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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.