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Anti-RACK1 Rabbit Monoclonal Antibody

Figure 1. Western blot analysis of RACK1 using anti-RACK1 antibody (M02145-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human MCF-7 whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat C6 whole cell lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse HEPA1-6 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-RACK1 antigen affinity purified monoclonal antibody (Catalog # M02145-1) at 1:500 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:1000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for RACK1 at approximately 35 kDa. The expected band size for RACK1 is at 35 kDa.
Figure 2. IF analysis of RACK1 using anti-RACK1 antibody (M02145-1) and anti-Beta Tubulin antibody (M01857-3).
RACK1 was detected in immunocytochemical section of A549 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated at 1:50 with rabbit anti-RACK1 Antibody (M02145-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF, ICC, FC
Primary Accession	P63244
Host	Rabbit
Isotype	IgG
Reactivity	Rat, Human, Mouse
Clonality	Monoclonal
Format	Liquid
Description	Anti-RACK1 Rabbit Monoclonal Antibody . Tested in WB, IHC, ICC/IF, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.

Additional Information
Gene ID	10399
Other Names	Small ribosomal subunit protein RACK1, Cell proliferation-inducing gene 21 protein, Guanine nucleotide-binding protein subunit beta-2-like 1, Guanine nucleotide-binding protein subunit beta-like protein 12.3, Human lung cancer oncogene 7 protein, HLC-7, Receptor for activated C kinase, Receptor of activated protein C kinase 1, Small ribosomal subunit protein RACK1, N-terminally processed, Guanine nucleotide-binding protein subunit beta-2-like 1, N-terminally processed, Receptor of activated protein C kinase 1, N-terminally processed, RACK1 (HGNC:4399), GNB2L1
Calculated MW	35077 Da
Application Details	WB 1:500-1:2000 IHC 1:50-1:200 ICC/IF 1:50-1:200 FC 1:50
Source	Eukaryota
Contents	Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clone Names	Clone: 20R31
Immunogen	A synthesized peptide derived from human RACK1
Purification	Affinity-chromatography
Storage	Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	RACK1 (HGNC:4399)
Synonyms	GNB2L1
Function	Scaffolding protein involved in the recruitment, assembly and/or regulation of a variety of signaling molecules. Interacts with a wide variety of proteins and plays a role in many cellular processes. Component of the 40S ribosomal subunit involved in translational repression (PubMed:23636399). Involved in the initiation of the ribosome quality control (RQC), a pathway that takes place when a ribosome has stalled during translation, by promoting ubiquitination of a subset of 40S ribosomal subunits (PubMed:28132843). Binds to and stabilizes activated protein kinase C (PKC), increasing PKC-mediated phosphorylation. May recruit activated PKC to the ribosome, leading to phosphorylation of EIF6. Inhibits the activity of SRC kinases including SRC, LCK and YES1. Inhibits cell growth by prolonging the G0/G1 phase of the cell cycle. Enhances phosphorylation of BMAL1 by PRKCA and inhibits transcriptional activity of the BMAL1-CLOCK heterodimer. Facilitates ligand-independent nuclear translocation of AR following PKC activation, represses AR transactivation activity and is required for phosphorylation of AR by SRC. Modulates IGF1R-dependent integrin signaling and promotes cell spreading and contact with the extracellular matrix. Involved in PKC-dependent translocation of ADAM12 to the cell membrane. Promotes the ubiquitination and proteasome- mediated degradation of proteins such as CLEC1B and HIF1A. Required for VANGL2 membrane localization, inhibits Wnt signaling, and regulates cellular polarization and oriented cell division during gastrulation. Required for PTK2/FAK1 phosphorylation and dephosphorylation. Regulates internalization of the muscarinic receptor CHRM2. Promotes apoptosis by increasing oligomerization of BAX and disrupting the interaction of BAX with the anti-apoptotic factor BCL2L. Inhibits TRPM6 channel activity. Regulates cell surface expression of some GPCRs such as TBXA2R. Plays a role in regulation of FLT1-mediated cell migration. Involved in the transport of ABCB4 from the Golgi to the apical bile canalicular membrane (PubMed:19674157). Promotes migration of breast carcinoma cells by binding to and activating RHOA (PubMed:20499158). Acts as an adapter for the dephosphorylation and inactivation of AKT1 by promoting recruitment of PP2A phosphatase to AKT1 (By similarity).
Cellular Location	Cell membrane; Peripheral membrane protein. Cytoplasm. Cytoplasm, perinuclear region. Nucleus. Perikaryon {ECO:0000250\|UniProtKB:P68040}. Cell projection, dendrite {ECO:0000250\|UniProtKB:P68040}. Cell projection, phagocytic cup. Note=Recruited to the plasma membrane through interaction with KRT1 which binds to membrane-bound ITGB1 (PubMed:17956333). Also associated with the membrane in oncogene- transformed cells (PubMed:11884618). PKC activation induces translocation from the perinuclear region to the cell periphery (PubMed:11279199). In the brain, detected mainly in cell bodies and dendrites with little expression in axonal fibers or nuclei (By similarity). Localized to phagocytic cups following infection by Y.pestis (PubMed:21347310). {ECO:0000250\|UniProtKB:P68040, ECO:0000269\|PubMed:11279199, ECO:0000269\|PubMed:11884618, ECO:0000269\|PubMed:17956333, ECO:0000269\|PubMed:21347310}
Tissue Location	In the liver, expressed at higher levels in activated hepatic stellate cells than in hepatocytes or Kupffer cells Up-regulated in hepatocellular carcinomas and in the adjacent non-tumor liver tissue.