ET7110-88

BAT3 Recombinant Rabbit Monoclonal Antibody [JE54-90]

cat.: ET7110-88

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JE54-90

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 119 kDa.
Isotype:	IgG
Immunogen:	Synthetic peptide within Human BAT3 aa 1,083-1,132 / 1,132.
Positive control:	Rat testis tissue lysate, 293 cell lysate, HepG2 cell lysate, rat brain tissue lysate, mouse testis tissue lysate, mouse hippocampus tissue lysate, THP-1 cell lysate, A549, HepG2, rat testis tissue, human lung tissue, human lung carcinoma tissue,SW620.
Subcellular location:	Extracellular exosome, cytosol, nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:500-1:2,000 1:50-1:100 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P46379 Human \| Q9Z1R2 Mouse \| Q6MG49 Rat
Alternative names:	2410045D21Rik AA408914 BAG 6 BAG family molecular chaperone regulator 6 BAG-6 BAG6 BAG6_HUMAN BAT 3 BAT3 BCL2-associated athanogene 6 D17H6S52E D6S52E G3 HLA B associated transcript 3 HLA-B associated transcript 3 HLA-B associated transcript-3 HLA-B-associated transcript 3 large proline rich protein BAG6 Large proline rich protein BAT3 Large proline-rich protein BAG6 large proline-rich protein BAT3 Protein G3 Protein Scythe Scythe Scythe, homolog of Xenopus

Images

Fig1: Western blot analysis of BAT3 on different lysates with Rabbit anti-BAT3 antibody (ET7110-88) at 1/500 dilution.

Lane 1: Rat testis tissue lysate
Lane 2: 293 cell lysate
Lane 3: HepG2 cell lysate
Lane 4: Rat brain tissue lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 119 kDa
Observed band size: 150 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 (ET7110-88) 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 1:5,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of BAT3 on different lysates with Rabbit anti-BAT3 antibody (ET7110-88) at 1/500 dilution.

Lane 1: Mouse testis tissue lysate
Lane 2: Mouse hippocampus tissue lysate
Lane 3: THP-1 cell lysate(10 µg/Lane.)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 119 kDa
Observed band size: 150 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 (ET7110-88) 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 1:5,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of A549 cells labeling BAT3 with Rabbit anti-BAT3 antibody (ET7110-88) 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-BAT3 antibody (ET7110-88) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 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 HepG2 cells labeling BAT3 with Rabbit anti-BAT3 antibody (ET7110-88) 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-BAT3 antibody (ET7110-88) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-BAT3 antibody (ET7110-88) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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 (ET7110-88) at 1/200 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-BAT3 antibody (ET7110-88) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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 (ET7110-88) at 1/200 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 lung carcinoma tissue with Rabbit anti-BAT3 antibody (ET7110-88) at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 6.0) 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 (ET7110-88) 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 BAT3 was done on SW620 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-88, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated 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”.