ET1701-17

Sumo 2+3 Recombinant Rabbit Monoclonal Antibody [JJ087-04]

cat.: ET1701-17

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P
Clonality:	Monoclonal
Clone number:	JJ087-04

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 12 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Sumo 3 aa 11-60 / 103.
Positive control:	HeLa cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Human lung tissue lysate, Mouse kidney tissue lysate, HeLa, NIH/3T3, human kidney tissue, mouse kidney tissue, rat kidney tissue, mouse large intestine tissue, rat skin tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P	1:500-1:1,000 1:50-1:200 1:50-1:200 1:50-1:1,000
Uniprot #:	SwissProt: P55854 Human \| P61956 Human \| P61957 Mouse \| Q9Z172 Mouse \| P61959 Rat \| Q5XIF4 Rat
Alternative names:	HSMT3 MGC117191 OTTHUMP00000115275 OTTHUMP00000115276 OTTHUMP00000115277 Sentrin 2 Small ubiquitin like modifier 2 Small ubiquitin related modifier 2 small ubiquitin-like modifier 3 small ubiquitin-related modifier 3 SMT3 homolog 1 SMT3 homolog 2 SMT3 suppressor of mif two 3 homolog 1 SMT3 suppressor of mif two 3 homolog 2 (S. cerevisiae) SMT3 suppressor of mif two 3 homolog 2 SMT3 suppressor of mif two 3 homolog 3 (S. cerevisiae) SMT3 suppressor of mif two 3 homolog 3 SMT3A SMT3B SMT3H1 SMT3H2 Sumo2 Sumo3 Ubiquitin like protein SMT3A Ubiquitin like protein SMT3B

Images

Fig1: Western blot analysis of Sumo 2+3 on different lysates with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
Lane 3: PC-12 cell lysate (20 µg/Lane)
Lane 4: Human lung tissue lysate (40 µg/Lane)
Lane 5: Mouse kidney tissue lysate (40 µg/Lane)

Predicted band size: 12 kDa
Observed band size: 16 kDa

Exposure time: 2 minutes; ECL: K1802;

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 (ET1701-17) 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: Immunocytochemistry analysis of HeLa cells labeling Sumo 2+3 with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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-Sumo 2+3 antibody (ET1701-17) 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.

	Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling Sumo 2+3 with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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-Sumo 2+3 antibody (ET1701-17) 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: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/200 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 (ET1701-17) at 1/200 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 mouse kidney tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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 (ET1701-17) 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 rat kidney tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) 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 (ET1701-17) 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: Immunohistochemical analysis of paraffin-embedded mouse large intestine tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/400 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 (ET1701-17) at 1/400 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-Sumo 2+3 antibody (ET1701-17) at 1/400 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 (ET1701-17) at 1/400 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”.