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RACK1 Recombinant Rabbit Monoclonal Antibody [JG40-26]

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Specification

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Catalog# ET7109-04

RACK1 Recombinant Rabbit Monoclonal Antibody [JG40-26]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Predicted reactivity

  • Zebrafish

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

RACK1 Recombinant Rabbit Monoclonal Antibody [JG40-26]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human RACK1 aa 190-317 / 317.

Species Reactivity

Human, Mouse, Rat (Predicted: Zebrafish)

Predicted species support after-sales service

Validated Applications

WB, IHC-P, FC

Molecular Weight

Predicted band size: 35 kDa

Positive Control

Daudi cell lysate, C2C12 cell lysate, C6 cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, SH-SY5Y cell lysate, human liver tissue, mouse liver tissue, rat liver tissue, SH-SY5Y.

Conjugation

unconjugated

Clone Number

JG40-26

RRID

AB_3070894

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:50-1:100

Target

Function

Members of the protein kinase C (PKC) family play a key regulatory role in a variety of cellular functions, including cell growth and differentiation, gene expression, hormone secretion and membrane function. Receptor for activated C kinases, termed RACKs, are intracellular receptors for activated PKC that may be involved in the activation-induced translocation of PKC. RACK1 (receptor for activated C kinase 1) is a 317 amino acid G protein b subunit-like protein that functions as a RACK and inhibits the activity of Src tyrosine kinases. In response to PKC activation, the intracellular localization of RACK1 and PKC bII changes, and RACK1 and PKC bII co-localize to the same sites. RACK1 is therefore thought to be a shuttling protein for PKC bII. A deficit in RACK1 may be associated with impaired PKC activation in the aging brain. The RACK1 gene is conserved in chimpanzee, dog, cow, mouse, rat, chicken, zebrafish, fruit fly, mosquito, C.elegans, S.pombe, S.cerevisiae, K.lactis, E.gossypii, M.grisea, N.crassa, A.thaliana, rice and P.falciparum, and maps to human chromosome 5q35.3.

Background References

1. Chang B Y et al. RACK1, a receptor for activated C kinase and a homolog of the beta subunit of G proteins, inhibits activity of src tyrosine kinases and growth of NIH 3T3 cells. Mol Cell Biol 18:3245-3256 (1998).

2. Hermanto U et al. RACK1, an insulin-like growth factor I (IGF-I) receptor-interacting protein, modulates IGF-I-dependent integrin signaling and promotes cell spreading and contact with extracellular matrix. Mol Cell Biol 22:2345-2365 (2002).

Sequence Similarity

Belongs to the WD repeat G protein beta family. Ribosomal protein RACK1 subfamily.

Tissue Specificity

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.

Post-translational Modification

Phosphorylated on Tyr-228 and/or Tyr-246 by SRC. This is required for binding to SRC.; (Microbial infection) Phosphorylated by vaccinia virus B1 kinase on serine and threonine residues; this phosphorylation remodels the ribosome properties, favoring the viral mRNA translation.

Subcellular Location

Cytoplasm. Cell membrane. Nucleus.

UNIPROT

SwissProt: P63244 Human

SwissProt: P63245 Rat

Synonyms

Cell proliferation-inducing gene 21 protein antibody

GBLP_HUMAN antibody

Gnb2-rs1 antibody

Gnb2l1 antibody

Guanine nucleotide binding protein (G protein) beta polypeptide 2 like 1 antibody

Guanine nucleotide binding protein beta polypeptide 2 like 1 antibody

Guanine nucleotide binding protein beta subunit 2 like 1 antibody

Guanine nucleotide binding protein beta subunit like protein 12.3 antibody

Guanine nucleotide binding protein subunit beta 2 like 1 antibody

Guanine nucleotide binding protein subunit beta like protein 12.3 antibody

Expand

Images

  • Western blot analysis of RACK1 on different lysates with Rabbit anti-RACK1 antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/1,000 dilution.<br /><br />Lane 1: Daudi cell lysate (20 µg/Lane)<br />Lane 2: C2C12 cell lysate (20 µg/Lane)<br />Lane 3: C6 cell lysate (20 µg/Lane)<br />Lane 4: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 5: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 35 kDa<br />Observed band size: 32 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of RACK1 on different lysates with Rabbit anti-RACK1 antibody (ET7109-04) at 1/1,000 dilution.

    Lane 1: Daudi cell lysate (20 µg/Lane)
    Lane 2: C2C12 cell lysate (20 µg/Lane)
    Lane 3: C6 cell lysate (20 µg/Lane)
    Lane 4: Mouse liver tissue lysate (40 µg/Lane)
    Lane 5: Rat liver tissue lysate (40 µg/Lane)

    Predicted band size: 35 kDa
    Observed band size: 32 kDa

    Exposure time: 25 seconds; ECL: K1801;

    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 (ET7109-04) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of RACK1 on different lysates with Rabbit anti-RACK1 antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/1,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate<br />Lane 2: Daudi cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 35 kDa<br />Observed band size: 35 kDa<br /><br />Exposure time: 43 seconds;<br /><br />12% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of RACK1 on different lysates with Rabbit anti-RACK1 antibody (ET7109-04) at 1/1,000 dilution.

    Lane 1: SH-SY5Y cell lysate
    Lane 2: Daudi cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 35 kDa
    Observed band size: 35 kDa

    Exposure time: 43 seconds;

    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 (ET7109-04) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-RACK1 antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/200 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.

    Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-RACK1 antibody (ET7109-04) at 1/200 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 (ET7109-04) at 1/200 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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-RACK1 antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-RACK1 antibody (ET7109-04) 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 (ET7109-04) 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.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-RACK1 antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/200 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>) at 1/200 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.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-RACK1 antibody (ET7109-04) at 1/200 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 (ET7109-04) at 1/200 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.

  • Flow cytometric analysis of RACK1 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7109-04" style="font-weight: bold;text-decoration: underline;">ET7109-04</a>, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-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; yellow).

    Flow cytometric analysis of RACK1 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7109-04, 1/50) (purple). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-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; yellow).

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Citation