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PKC beta 2 Recombinant Rabbit Monoclonal Antibody [ST48-06]

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Specification

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Catalog# ET1609-44

PKC beta 2 Recombinant Rabbit Monoclonal Antibody [ST48-06]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

PKC beta 2 Recombinant Rabbit Monoclonal Antibody [ST48-06]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human PKC beta 2 aa 615-660 / 671.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

Molecular Weight

Predicted band size: 77 kDa

Positive Control

K-562 cell lysate, Raji cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, human spleen tissue, mouse spleen tissue, rat spleen tissue, SH-SY5Y, Hela.

Conjugation

unconjugated

Clone Number

ST48-06

RRID

AB_3069854

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

  • 1:500

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

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. PKCs were originally identified as serine/threonine protein kinases whose activity was dependent on calcium and phospholipids. Diacylglycerols (DAG) and tumor promoting phorbol esters bind to and activate PKC. PKCs can be subdivided into at least two major classes including conventional PKC isoforms (α, βI, βII and γ) and novel PKC isoforms. Patterns of expression for each PKC isoform differ among tissues and PKC family members exhibit clear differences in their cofactor dependencies. For instance, the kinase activities of nPKC δ and é are independent of Ca2+. On the other hand, nPKC δ and ε, as well as all of the cPKC members, possess phorbol ester-binding activities and kinase activities.

Background References

1. Lin G et al. Complex regulation of PKC 2 and PDK-1/AKT by ROCK2 in diabetic heart. PLoS One 9:e86520 (2014).

2. Qu C et al. Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-kB activation in diffuse large B-cell lymphoma. Blood 121:4718-28 (2013).

Sequence Similarity

Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.

Post-translational Modification

Phosphorylation on Thr-500 within the activation loop renders it competent to autophosphorylate. Subsequent autophosphorylation of Thr-642 maintains catalytic competence, and autophosphorylation on Ser-661 appears to release the kinase into the cytosol. Autophosphorylation on other sites i.e. in the N-terminal and hinge regions have no effect on enzyme activity. Phosphorylation at Tyr-662 by SYK induces binding with GRB2 and contributes to the activation of MAPK/ERK signaling cascade (By similarity).

Subcellular Location

Nucleus, Cytoplasm, Membrane.

UNIPROT

SwissProt: P05771 Human

SwissProt: P68404 Mouse

SwissProt: P68403 Rat

Synonyms

KPCB_HUMAN antibody

PKC Beta antibody

PKC-B antibody

PKC-beta antibody

PKCB antibody

Prkcb antibody

PRKCB II antibody

PRKCB2 antibody

Protein kinase C beta antibody

Protein kinase C beta type antibody

Images

  • Western blot analysis of PKC beta 2 on different lysates with Rabbit anti-PKC beta 2 antibody (<a href="/products/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</a>) at 1/500 dilution.<br /><br />Lane 1: K-562 cell lysate (20 µg/Lane)<br />Lane 2: Raji cell lysate (20 µg/Lane)<br />Lane 3: Mouse spleen tissue lysate (40 µg/Lane)<br />Lane 4: Rat spleen tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 77 kDa<br />Observed band size: 77 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<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/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</a>) at 1/500 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 PKC beta 2 on different lysates with Rabbit anti-PKC beta 2 antibody (ET1609-44) at 1/500 dilution.

    Lane 1: K-562 cell lysate (20 µg/Lane)
    Lane 2: Raji cell lysate (20 µg/Lane)
    Lane 3: Mouse spleen tissue lysate (40 µg/Lane)
    Lane 4: Rat spleen tissue lysate (40 µg/Lane)

    Predicted band size: 77 kDa
    Observed band size: 77 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 (ET1609-44) at 1/500 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 spleen tissue with Rabbit anti-PKC beta 2 antibody (<a href="/products/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</a>) at 1/50 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 spleen tissue with Rabbit anti-PKC beta 2 antibody (ET1609-44) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1609-44) at 1/50 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 spleen tissue with Rabbit anti-PKC beta 2 antibody (<a href="/products/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</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 spleen tissue with Rabbit anti-PKC beta 2 antibody (ET1609-44) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1609-44) 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 spleen tissue with Rabbit anti-PKC beta 2 antibody (<a href="/products/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</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/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</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 spleen tissue with Rabbit anti-PKC beta 2 antibody (ET1609-44) 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 (ET1609-44) 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.

  • ICC staining of PKC beta 2 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of PKC beta 2 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1609-44, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Flow cytometric analysis of PKC beta 2 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1609-44" style="font-weight: bold;text-decoration: underline;">ET1609-44</a>, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated 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; red).

    Flow cytometric analysis of PKC beta 2 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-44, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated 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; red).

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Citation