ET1706-34

Drosha Recombinant Rabbit Monoclonal Antibody [JU33-01]

cat.: ET1706-34

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JU33-01

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:	159 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Drosha aa 30-79 / 1,374.
Positive control:	Hela cell lysate, SiHa cell lysate, K562 cell lysates, A549, Hela, LOVO, human kidney tissue, human placenta tissue, Jurkat.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:500-1:1,000 1:50-1:200 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: Q9NRR4 Human
Alternative names:	DROSHA Drosha double stranded RNA specific endoribonuclease Drosha ribonuclease type III Etohi2 HSA242976 Nuclear RNase III Drosha p241 Protein Drosha Putative protein p241 which interacts with transcription factor Sp1 Putative ribonuclease III RANSE3L Ribonuclease 3 Ribonuclease III Ribonuclease III nuclear Ribonuclease type III nuclear RibonucleaseIII RN 3 RN3 RNase 3 RNase III RNase3 RNASE3L RNaseIII RNASEN RNC_HUMAN

Images

Fig1: Western blot analysis of Drosha on different lysates with Rabbit anti-Drosha antibody (ET1706-34) at 1/500 dilution.

Lane 1: Hela cell lysate
Lane 2: SiHa cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 159 kDa
Observed band size: 159 kDa

Exposure time: 2 minutes;

6% 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 (ET1706-34) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 200,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of Drosha on K562 cell lysates with Rabbit anti-Drosha antibody (ET1706-34) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 159 kDa
Observed band size: 159 kDa

Exposure time: 2 minutes;

6% 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 (ET1706-34) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of A549 cells labeling Drosha with Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, 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 Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
	Fig4: Immunocytochemistry analysis of Hela cells labeling Drosha with Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, 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 Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
	Fig5: Immunocytochemistry analysis of LOVO cells labeling Drosha with Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, 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 Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

	Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1706-34) at 1/50 dilution for 0.5 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 human placenta tissue with Rabbit anti-Drosha antibody (ET1706-34) at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1706-34) at 1/50 dilution for 0.5 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: Flow cytometric analysis of Drosha was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1706-34, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.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”.