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p70 S6 Kinase Recombinant Rabbit Monoclonal Antibody [JE03-20]

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Specification

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Catalog# HA722520

p70 S6 Kinase Recombinant Rabbit Monoclonal Antibody [JE03-20]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

p70 S6 Kinase Recombinant Rabbit Monoclonal Antibody [JE03-20]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human p70 S6 Kinase aa 1-50 / 525.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 59 kDa

Positive Control

MCF7 cell lysate, A549 cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, human brain tissue, mouse brain tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

JE03-20

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. 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

  • IHC-P

  • 1:200

  • IP

  • 1-2μg/sample

Target

Function

Serine/threonine-protein kinase that acts downstream of mTOR signaling in response to growth factors and nutrients to promote cell proliferation, cell growth and cell cycle progression. Upon mitogenic stimulation, phosphorylation by the mechanistic target of rapamycin complex 1 (mTORC1) leads to dissociation from the EIF3 complex and activation. The active form then phosphorylates and activates several substrates in the pre-initiation complex, including the EIF2B complex and the cap-binding complex component EIF4B. Also controls translation initiation by phosphorylating a negative regulator of EIF4A, PDCD4, targeting it for ubiquitination and subsequent proteolysis. Promotes initiation of the pioneer round of protein synthesis by phosphorylating POLDIP3/SKAR. In response to IGF1, activates translation elongation by phosphorylating EEF2 kinase (EEF2K), which leads to its inhibition and thus activation of EEF2. Also plays a role in feedback regulation of mTORC2 by mTORC1 by phosphorylating RICTOR, resulting in the inhibition of mTORC2 and AKT1 signaling.

Background References

1. Wang X., Li W., Williams M., Terada N., Alessi D.R., Proud C.G. Regulation of elongation factor 2 kinase by p90(RSK1) and p70 S6 kinase. EMBO J. 20:4370-4379 (2001)

2. Fleckenstein D.S., Dirks W.G., Drexler H.G., Quentmeier H. Tumor necrosis factor receptor-associated factor (TRAF) 4 is a new binding partner for the p70S6 serine/threonine kinase. Leuk. Res. 27:687-694 (2003)

Subcellular Location

Cytoplasm. Mitochondrion. Mitochondrion outer membrane.

UNIPROT

SwissProt: P23443 Human

SwissProt: Q8BSK8 Mouse

SwissProt: P67999 Rat

Synonyms

70 kDa ribosomal protein S6 kinase 1 antibody

KS6B1_HUMAN antibody

p70 alpha antibody

P70 beta 1 antibody

p70 ribosomal S6 kinase alpha antibody

p70 ribosomal S6 kinase beta 1 antibody

p70 S6 kinase alpha antibody

P70 S6 Kinase antibody

p70 S6 kinase, alpha 1 antibody

p70 S6 kinase, alpha 2 antibody

Expand

Images

  • Western blot analysis of p70 S6 Kinase on different lysates with Rabbit anti-p70 S6 Kinase antibody (<a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</a>) at 1/1,000 dilution.<br /><br />Lane 1: MCF7 cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: Mouse brain tissue lysate<br />Lane 7: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 59 kDa<br />Observed band size: 59 kDa<br /><br />Exposure time: 10 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/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</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 p70 S6 Kinase on different lysates with Rabbit anti-p70 S6 Kinase antibody (HA722520) at 1/1,000 dilution.

    Lane 1: MCF7 cell lysate
    Lane 2: A549 cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: C6 cell lysate
    Lane 6: Mouse brain tissue lysate
    Lane 7: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 59 kDa
    Observed band size: 59 kDa

    Exposure time: 10 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 (HA722520) 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.

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-p70 S6 Kinase antibody (<a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</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/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</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 brain tissue with Rabbit anti-p70 S6 Kinase antibody (HA722520) 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 (HA722520) 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 brain tissue with Rabbit anti-p70 S6 Kinase antibody (<a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</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/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</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 mouse brain tissue with Rabbit anti-p70 S6 Kinase antibody (HA722520) 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 (HA722520) 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 brain tissue with Rabbit anti-p70 S6 Kinase antibody (<a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</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/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</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 brain tissue with Rabbit anti-p70 S6 Kinase antibody (HA722520) 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 (HA722520) 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.

  • p70 S6 Kinase was immunoprecipitated from 0.2 mg C6 cell lysate with <a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (<a href="/products/NBI01H" style="font-weight: bold;text-decoration: underline;">NBI01H</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: C6 cell lysate (input)<br />Lane 2: <a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</a> IP in C6 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722520" style="font-weight: bold;text-decoration: underline;">HA722520</a> in C6 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 20 seconds; ECL: K1801

    p70 S6 Kinase was immunoprecipitated from 0.2 mg C6 cell lysate with HA722520 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722520 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: C6 cell lysate (input)
    Lane 2: HA722520 IP in C6 cell lysate
    Lane 3: Rabbit IgG instead of HA722520 in C6 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 20 seconds; ECL: K1801

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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