HA721992

LEF1 Recombinant Rabbit Monoclonal Antibody [JE52-28]

cat.: HA721992

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	JE52-28

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: 44 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human LEF1 aa 121-170 / 399.
Positive control:	Jurkat cell lysate, mouse thymus tissue lysate, rat thymus tissue lysate, Jurkat, human lymph node tissue, human thymus tissue, human tonsil tissue, rat thymus tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P	1:2,000 1:500 1:500-1:2,000
Uniprot #:	SwissProt: Q9UJU2 Human \| P27782 Mouse \| Q9QXN1 Rat
Alternative names:	DKFZp586H0919 FLJ46390 LEF 1 LEF-1 Lef1 LEF1_HUMAN Lymphoid enhancer binding factor 1 Lymphoid enhancer-binding factor 1 T cell specific transcription factor 1 alpha T cell-specific transcription factor 1-alpha TCF 1 alpha TCF1 alpha TCF1-alpha TCF10 TCF1alpha TCF7L3 Transcription factor T cell specific 1 alpha

Images

Fig1: Western blot analysis of LEF1 on different lysates with Rabbit anti-LEF1 antibody (HA721992) at 1/2,000 dilution.

Lane 1: Jurkat cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (negative) (20 µg/Lane)
Lane 3: Mouse thymus tissue lysate (40 µg/Lane)
Lane 4: Rat thymus tissue lysate (40 µg/Lane)

Predicted band size: 44 kDa
Observed band size: 35-55 kDa

Exposure time: 3 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 (HA721992) at 1/2,000 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.

Fig2: Western blot analysis of LEF1 on different lysates with Rabbit anti-LEF1 antibody (HA721992) at 1/2,000 dilution.

Lane 1: HAP1-parental cell lysate (10 µg/Lane)
Lane 2: HAP1-LEF1 KD cell lysate (10 µg/Lane)

Predicted band size: 44 kDa
Observed band size: 35-55 kDa

Exposure time: 3 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 (HA721992) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of Jurkat (positive) and HeLa (negative) labeling LEF1 with Rabbit anti-LEF1 antibody (HA721992) at 1/500 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-LEF1 antibody (HA721992) at 1/500 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human lymph node tissue with Rabbit anti-LEF1 antibody (HA721992) 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 (HA721992) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human thymus tissue with Rabbit anti-LEF1 antibody (HA721992) 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 (HA721992) 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.

Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-LEF1 antibody (HA721992) at 1/500 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 (HA721992) at 1/500 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.

Fig7: Immunohistochemical analysis of paraffin-embedded rat thymus tissue with Rabbit anti-LEF1 antibody (HA721992) 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 (HA721992) 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.