HA721815

HUWE1 Recombinant Rabbit Monoclonal Antibody [PSH02-38]

cat.: HA721815

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	PSH02-38

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: 482.7 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within rat HUWE1 aa 2,701-2,750 / 4,378 (A0A8I6A634).
Positive control:	PC-12 cell lysates, human brain tissue, human kidney tissue, mouse kidney tissue, rat kidney tissue, HeLa, PC-12.
Subcellular location:	Nucleus
Recommended Dilutions: WB IHC-P IF-Cell FC	1:1,000 1:1,000 1:100 1:1,000
Uniprot #:	SwissProt: A0A8I6A634 Rat \| A0A994J483 Human \| A0A6P7QKZ7 Mouse
Alternative names:	ARF binding protein 1 ARF BP1 ARF-binding protein 1 ARF-BP1 ARFBP1 BJ-HCC-24 tumor antigen E3 ubiquitin protein ligase HUWE1 E3 ubiquitin-protein ligase HUWE1 HECT HECT domain protein LASU1 HECT UBA and WWE domain containing protein 1 HectH9 Homologous to E6AP carboxyl terminus homologous protein 9 Huwe1 HUWE1_HUMAN Ib772 Large structure of UREB1 LASU1 Mcl 1 ubiquitin ligase E3 Mcl-1 ubiquitin ligase E3 Mule UBA and WWE domain-containing protein 1 Upstream regulatory element-binding protein 1 URE-B1 URE-binding protein 1 UREB1

Images

	Fig1: Western blot analysis of HUWE1 on PC-12 cell lysates with Rabbit anti-HUWE1 antibody (HA721815) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 482.7 kDa Observed band size: 482.7 kDa Exposure time: 5 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 (HA721815) 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.
	Fig2: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-HUWE1 antibody (HA721815) 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 (HA721815) 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 kidney tissue with Rabbit anti-HUWE1 antibody (HA721815) 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 (HA721815) 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 mouse kidney tissue with Rabbit anti-HUWE1 antibody (HA721815) 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 (HA721815) 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 rat kidney tissue with Rabbit anti-HUWE1 antibody (HA721815) 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 (HA721815) 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: Immunocytochemistry analysis of HeLa cells labeling HUWE1 with Rabbit anti-HUWE1 antibody (HA721815) at 1/100 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-HUWE1 antibody (HA721815) 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 (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.

Fig7: Immunocytochemistry analysis of PC-12 cells labeling HUWE1 with Rabbit anti-HUWE1 antibody (HA721815) at 1/100 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-HUWE1 antibody (HA721815) 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 (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.

Fig8: Flow cytometric analysis of HeLa cells labeling HUWE1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721815, 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”.