HA720004

Ube4A Recombinant Rabbit Monoclonal Antibody [JE56-68]

cat.: HA720004

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JE56-68

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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: 123 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human Ube4A aa 1-100.
Positive control:	293T cell lysate, HepG2 cell lysate, PC-12 cell lysate, Rat brain tissue lysate, Mouse brain tissue lysate, Mouse kidney tissue lysate, HepG2, PC-12, human small intestine tissue, 293T.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:5,000 1:50-1:100 1:50-1:200 1:1,000
Uniprot #:	SwissProt: Q14139 Human \| E9Q735 Mouse \| Q6P7A2 Rat
Alternative names:	E4 KIAA0126 MGC133315 UBE4A UBE4A_HUMAN Ubiquitin conjugation factor E4 A ubiquitination factor E4A (UFD2 homolog yeast) Ubiquitination factor E4A UBOX2 UFD2 UFD2 homolog UFD2b

Images

Fig1: Western blot analysis of Ube4A on different lysates with Rabbit anti-Ube4A antibody (HA720004) at 1/5,000 dilution.

Lane 1: 293T cell lysate (10 µg/Lane)
Lane 2: HepG2 cell lysate (10 µg/Lane)
Lane 3: PC-12 cell lysate (10 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 123 kDa
Observed band size: 123 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 (HA720004) at 1/5,000 dilution was used in primary antibody dilution (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.

Fig2: Western blot analysis of Ube4A on different lysates with Rabbit anti-Ube4A antibody (HA720004) at 1/5,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Mouse kidney tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 123 kDa
Observed band size: 123 kDa

Exposure time: 59 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 (HA720004) at 1/5,000 dilution was used in primary antibody dilution (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 HepG2 cells labeling Ube4A with Rabbit anti-Ube4A antibody (HA720004) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Ube4A antibody (HA720004) at 1/100 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 (HA601187, 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: Immunocytochemistry analysis of PC-12 cells labeling Ube4A with Rabbit anti-Ube4A antibody (HA720004) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Ube4A antibody (HA720004) at 1/100 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 (HA601187, 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-Ube4A antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA720004, 1/50) for 30 minutes 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: Flow cytometric analysis of 293T cells labeling Ube4A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA720004, 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).

Fig7: Flow cytometric analysis of PC-12 cells labeling Ube4A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA720004, 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).

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