RAC1/2/3 Rabbit Polyclonal Antibody
Catalog# HA500424
RAC1/2/3 Rabbit Polyclonal Antibody
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WB
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IHC-P
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IF-Cell
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
RAC1/2/3 Rabbit Polyclonal Antibody
Antibody Type
Rabbit Polyclonal Antibody
Immunogen
Recombinant protein within Human RAC3 aa 23-192 / 192.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Cell, FC
Target Molecular Weight
Predicted band size: 21 kDa
Positive Control
Rat brain tissue lysate, Human brain tissue lysate, Mouse brain tissue lysate, Mouse cerebellum tissue lysate, HeLa, PC-12, rat skin tissue.
Conjugation
unconjugated
RRID
Product Features
Form
Liquid
Concentration
1.958 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Immunogen affinity purified.
Application Dilution
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WB
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1:5000
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IHC-P
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1:600
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IF-Cell
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1:100
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FC
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1:1,000
Target
Function
Rac1 function: Plasma membrane-associated small GTPase which cycles between active GTP-bound and inactive GDP-bound states. In its active state, binds to a variety of effector proteins to regulate cellular responses such as secretory processes, phagocytosis of apoptotic cells, epithelial cell polarization and growth-factor induced formation of membrane ruffles. Rac2 fucntion: Plasma membrane-associated small GTPase which cycles between an active GTP-bound and inactive GDP-bound state. In active state binds to a variety of effector proteins to regulate cellular responses, such as secretory processes, phagocytose of apoptotic cells and epithelial cell polarization. Augments the production of reactive oxygen species (ROS) by NADPH oxidase. Enzyme regulation: Regulated by guanine nucleotide exchange factors (GEFs) which promote the exchange of bound GDP for free GTP, GTPase activating proteins (GAPs) which increase the GTP hydrolysis activity, and GDP dissociation inhibitors which inhibit the dissociation of the nucleotide from the GTPase. Rac3 function: Plasma membrane-associated small GTPase which cycles between an active GTP-bound and inactive GDP-bound state. In active state binds to a variety of effector proteins to regulate cellular responses, such as cell spreading and the formation of actin-based protusions including lamellipodia and membrane ruffles. Promotes cell adhesion and spreading on fibrinogen in a CIB1 and alpha-IIb/beta3 integrin-mediated manner.
Background References
1. Cheng C et al. RAC3 Promotes Proliferation, Migration and Invasion via PYCR1/JAK/STAT Signaling in Bladder Cancer. Front Mol Biosci. 2020 Aug
2. Chen M et al. Rac3 Expression and its Clinicopathological Significance in Patients With Bladder Cancer. Pathol Oncol Res. 2021
Subcellular Location
Cytoplasm, Cytoplasm, Cell membrane.
UNIPROT
Synonyms
Cell migration-inducing gene 5 protein antibody
EN-7 antibody
GX antibody
HSPC022 antibody
MIG5 antibody
p21-Rac1 antibody
p21-Rac2 antibody
p21-Rac3 antibody
RAC1 antibody
RAC2 antibody
ExpandImages
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Western blot analysis of RAC1/2/3 on mouse brain tissue lysate with Rabbit anti-RAC1/2/3 antibody (HA500424) at 1/5,000 dilution.
Lysates/proteins at 20 µg/Lane.
Exposure time: 59 seconds ; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA500424, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 21 kDa
Observed band size: 21 kDa -
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. -
Immunocytochemistry analysis of PC-12 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. -
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). -
Flow cytometric analysis of PC-12 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). -
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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