HA500424

RAC1/2/3 Rabbit Polyclonal Antibody

cat.: HA500424

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Polyclonal

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:	1.958ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 21 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human RAC3 aa 23-192 / 192.
Positive control:	Rat brain tissue lysate, Human brain tissue lysate, Mouse brain tissue lysate, Mouse cerebellum tissue lysate, HeLa, NIH/3T3, rat skin tissue.
Subcellular location:	Cytoplasm, Cytoplasm, Cell membrane.
Recommended Dilutions: WB IHC-P IF-Cell FC	1:500 1:600 1:100 1:1,000
Uniprot #:	SwissProt: P15153 Human \| P60763 Human \| P63000 Human \| P60764 Mouse \| P63001 Mouse \| Q05144 Mouse \| Q6RUV5 Rat \| Q5U1Y2 Rat \| M0R5T4 Rat
Alternative names:	Cell migration-inducing gene 5 protein EN-7 GX HSPC022 MIG5 p21-Rac1 p21-Rac2 p21-Rac3 RAC1 RAC2 RAC3 Ras-like protein TC25 ras-related C3 botulinum toxin substrate 1 (rho family, small GTP binding protein Rac1) Ras-related C3 botulinum toxin substrate 1 ras-related C3 botulinum toxin substrate 2 (rho family, small GTP binding protein Rac2) Ras-related C3 botulinum toxin substrate 2 ras-related C3 botulinum toxin substrate 3 (rho family, small GTP binding protein Rac3) Ras-related C3 botulinum toxin substrate 3 Small G protein TC25

Images

Fig1: Western blot analysis of RAC1/2/3 on different lysates with Rabbit anti-RAC1/2/3 antibody (HA500424) at 1/500 dilution.

Lane 1: Rat brain tissue lysate
Lane 2: Human brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 21 kDa
Observed band size: 21 kDa

Exposure time: 5 minutes;

15% 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 (HA500424) 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 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of RAC1/2/3 on different lysates with Rabbit anti-RAC1/2/3 antibody (HA500424) at 1/500 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Mouse cerebellum tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 21 kDa
Observed band size: 21 kDa

Exposure time: 1 minute;

12% 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 (HA500424) 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 1:300,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling RAC1/2/3 with Rabbit anti-RAC1/2/3 antibody (HA500424) 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-RAC1/2/3 antibody (HA500424) 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: Immunocytochemistry analysis of NIH/3T3 cells labeling RAC1/2/3 with Rabbit anti-RAC1/2/3 antibody (HA500424) 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-RAC1/2/3 antibody (HA500424) 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.
	Fig5: Flow cytometric analysis of HeLa cells labeling RAC1/2/3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA500424, 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).

Fig6: Flow cytometric analysis of NIH/3T3 cells labeling RAC1/2/3.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500424, 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).

Fig7: Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-RAC1/2/3 antibody (HA500424) at 1/600 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 (HA500424) at 1/600 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”.