HA721573

Rab3A Recombinant Rabbit Monoclonal Antibody [PSH01-08]

cat.: HA721573

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	PSH01-08

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: 25 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human Rab3A aa 121-170 / 220.
Positive control:	Human brain tissue lysate, mouse brain tissue lysate, rat brain tissue lysate, human cerebellum tissue, mouse cerebellum tissue, rat cerebellum tissue, SH-SY5Y, N2A, PC-12.
Subcellular location:	Cytoplasm, cytosol, Lysosome, Cytoplasmic vesicle, secretory vesicle, Cell projection, axon, Cell membrane, Presynapse, Postsynapse.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:5,000 1:1,000 1:100 1:500-1:1,000
Uniprot #:	SwissProt: P20336 Human \| P63011 Mouse \| P63012 Rat
Alternative names:	Rab 3A RAB 3A member RAS oncogene family Rab3a RAB3A member RAS oncogene family RAB3A_HUMAN RAS associated protein RAB 3A RAS associated protein RAB3A Ras related protein Rab 3A Ras related protein Rab3A Ras-related protein Rab-3A

Images

Fig1: Western blot analysis of Rab3A on different lysates with Rabbit anti-Rab3A antibody (HA721573) at 1/5,000 dilution.

Lane 1: Human brain tissue lysate
Lane 2: Human liver tissue lysate (negative)
Lane 3: Mouse brain tissue lysate
Lane 4: Mouse liver tissue lysate (negative)
Lane 5: Rat brain tissue lysate
Lane 6: Rat liver tissue lysate (negative)

Lysates/proteins at 40 µg/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 27 seconds;

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 (HA721573) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig2: Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-Rab3A antibody (HA721573) at 1/1,000 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 (HA721573) 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 mouse cerebellum tissue with Rabbit anti-Rab3A antibody (HA721573) at 1/1,000 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 (HA721573) 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 rat cerebellum tissue with Rabbit anti-Rab3A antibody (HA721573) at 1/1,000 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 (HA721573) 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: Immunocytochemistry analysis of SH-SY5Y cells labeling Rab3A with Rabbit anti-Rab3A antibody (HA721573) 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-Rab3A antibody (HA721573) 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.

	Fig6: Immunocytochemistry analysis of N2A cells labeling Rab3A with Rabbit anti-Rab3A antibody (HA721573) 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-Rab3A antibody (HA721573) 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 Rab3A with Rabbit anti-Rab3A antibody (HA721573) 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-Rab3A antibody (HA721573) 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 N2A cells labeling Rab3A. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721573, 1ug/ml) (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).

Fig9: Flow cytometric analysis of PC-12 cells labeling Rab3A.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721573, 1ug/ml) (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).

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