HA601130

UBA3 Recombinant Mouse Monoclonal Antibody

cat.: HA601130

Product Type:	Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell
Clonality:	Monoclonal

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 52 kDa
Isotype:	IgG1
Immunogen:	Recombinant protein within Human UBA3 aa 220-424 / 463.
Positive control:	HepG2 cell lysate, Hela cell lysate, 293T cell lysate, K-562 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, human lung tissue lysate, human breast tissue, human esophagus tissue, human lung carcinoma tissue, human lung tissue, human skin tissue, SiHa.
Subcellular location:	Cytosol, nucleus, cytoplasm.
Recommended Dilutions: WB IHC-P IF-Cell	1:1,000 1:1,000 1:200
Uniprot #:	SwissProt: Q8TBC4 Human \| Q8C878 Mouse \| Q99MI7 Rat
Alternative names:	DKFZp566J164 EC 6.3.2. hUba3 MGC22384 NEDD8 activating enzyme E1 catalytic subunit NEDD8 activating enzyme E1C Nedd8 activating enzyme hUba3 NEDD8-activating enzyme E1 catalytic subunit NEDD8-activating enzyme E1C uba3 UBA3 ubiquitin activating enzyme E1 homolog UBA3_HUMAN UBE1C Ubiquitin activating enzyme 3 Ubiquitin activating enzyme E1C Ubiquitin-activating enzyme 3 Ubiquitin-activating enzyme E1C Ubiquitin-like modifier-activating enzyme 3

Images

Fig1: Western blot analysis of UBA3 on different lysates with Mouse anti-UBA3 antibody (HA601130) at 1/500 dilution.

Lane 1: HepG2 cell lysate
Lane 2: Hela cell lysate
Lane 3: 293T cell lysate
Lane 4: K-562 cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: PC-12 cell lysate
Lane 8: Mouse brain tissue lysate
Lane 9: Human lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 52 kDa
Observed band size: 52 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 (HA601130) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

Fig2: Immunohistochemical analysis of paraffin-embedded human breast tissue with Mouse anti-UBA3 antibody (HA601130) at 1/1,000 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 (HA601130) at 1/1,000 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Mouse anti-UBA3 antibody (HA601130) at 1/1,000 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 (HA601130) at 1/1,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Mouse anti-UBA3 antibody (HA601130) at 1/1,000 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 (HA601130) at 1/1,000 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 lung tissue with Mouse anti-UBA3 antibody (HA601130) at 1/1,000 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 (HA601130) at 1/1,000 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 skin tissue with Mouse anti-UBA3 antibody (HA601130) at 1/1,000 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 (HA601130) at 1/1,000 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.

Fig7: Immunocytochemistry analysis of SiHa cells labeling UBA3 with Mouse anti-UBA3 antibody (HA601130) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Mouse anti-UBA3 antibody (HA601130) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

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