Skip to content

Plectin Recombinant Rabbit Monoclonal Antibody [SY29-04]

Usd: 385

Specification

Bulk Order Quote | Customization Request

Catalog# ET1607-80

Plectin Recombinant Rabbit Monoclonal Antibody [SY29-04]

Application

  • WB

  • IHC-P

  • IP

  • IF-Tissue

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Plectin Recombinant Rabbit Monoclonal Antibody [SY29-04]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Plectin aa 2-101 / 4,684.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 532 kDa

Positive Control

Human skeletal muscle tissue, mouse skeletal muscle tissue, rat skeletal muscle tissue, HeLa.

Conjugation

unconjugated

Clone Number

SY29-04

RRID

AB_3069783

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

  • 1:200-1:1,000

  • IP

  • 1-2μg/sample

  • IF-Tissue

  • 1:100-1:200

  • IF-Cell

  • 1:100

Target

Function

Plectin is a giant protein found in nearly all mammalian cells which acts as a link between the three main components of the cytoskeleton: actin microfilaments, microtubules and intermediate filaments. Plectin links the cytoskeleton to junctions found in the plasma membrane that structurally connect different cells. By holding these different networks together, plectin plays an important role in maintaining the mechanical integrity and viscoelastic properties of tissues. Plectin binds both Lamin B and vimentin, and this binding is regulated by a variety of protein kinases. Phosphorylation by PKA or PKC results in decreased binding to Lamin B, and phosphorylation by PKA enhances the plectin-vimentin interactions.

Background References

1. Kulkarni RM et al. Ron receptor signaling is protective against DSS-induced colitis in mice. Am J Physiol Gastrointest Liver Physiol 306:G1065-74 (2014).

2. Eisenberg JL et al. Plectin-containing, centrally localized focal adhesions exert traction forces in primary lung epithelial cells. J Cell Sci 126:3746-55 (2013).

Sequence Similarity

Belongs to the plakin or cytolinker family.

Tissue Specificity

Widely expressed with highest levels in muscle, heart, placenta and spinal cord.

Post-translational Modification

Phosphorylated by CDK1; regulates dissociation from intermediate filaments during mitosis.

Subcellular Location

Cell junction, Cytoplasm, Cytoskeleton.

UNIPROT

SwissProt: Q15149 Human

SwissProt: Q9QXS1 Mouse

SwissProt: P30427 Rat

Synonyms

EBS1 antibody

EBSO antibody

HD1 antibody

Hemidesmosomal protein 1 antibody

PCN antibody

pleC antibody

PLEC_HUMAN antibody

PLEC1 antibody

PLEC1b antibody

Plectin 1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Plectin on different lysates with Rabbit anti-Plectin antibody (<a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Plectin KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 532 kDa<br />Observed band size: 532 kDa<br /><br />Exposure time: 30 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/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a>) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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.

    ☑ Knockdown (KD)

    Western blot analysis of Plectin on different lysates with Rabbit anti-Plectin antibody (ET1607-80) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Plectin KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 532 kDa
    Observed band size: 532 kDa

    Exposure time: 30 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 (ET1607-80) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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 skeletal muscle tissue with Rabbit anti-Plectin antibody (<a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</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/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</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 human skeletal muscle tissue with Rabbit anti-Plectin antibody (ET1607-80) 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 (ET1607-80) 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 skeletal muscle tissue with Rabbit anti-Plectin antibody (<a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</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/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</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 skeletal muscle tissue with Rabbit anti-Plectin antibody (ET1607-80) 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 (ET1607-80) 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 skeletal muscle tissue with Rabbit anti-Plectin antibody (<a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</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/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</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 rat skeletal muscle tissue with Rabbit anti-Plectin antibody (ET1607-80) 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 (ET1607-80) 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.

  • Immunocytochemistry analysis of HeLa cells labeling Plectin with Rabbit anti-Plectin antibody (<a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Plectin antibody (<a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Plectin with Rabbit anti-Plectin antibody (ET1607-80) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Plectin antibody (ET1607-80) 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 (M1305-2, 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.

  • Plectin was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a> at 1/2,000 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: HeLa cell lysate (input)<br />Lane 2: <a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1607-80" style="font-weight: bold;text-decoration: underline;">ET1607-80</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 3 minutes; ECL: K1801

    Plectin was immunoprecipitated from 0.2 mg HeLa cell lysate with ET1607-80 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1607-80 at 1/2,000 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: HeLa cell lysate (input)
    Lane 2: ET1607-80 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of ET1607-80 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 3 minutes; ECL: K1801

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

Citation