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KLC1 Recombinant Rabbit Monoclonal Antibody [JE54-47]

Usd: 385

Specification

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Catalog# ET7110-72

KLC1 Recombinant Rabbit Monoclonal Antibody [JE54-47]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

KLC1 Recombinant Rabbit Monoclonal Antibody [JE54-47]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human KLC1 aa 461-573 / 573.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

65 kDa

Positive Control

Human brain tissue lysate, rat brain tissue lysate, Hela cell lysate, PC-12 cell lysate, MCF-7, RWPE-1, rat testis tissue, human kidney tissue, mouse brain tissue, SH-SY5Y.

Conjugation

unconjugated

Clone Number

JE54-47

RRID

AB_3071035

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

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:100

  • IHC-P

  • 1:50-1:500

  • FC

  • 1:50-1:100

Target

Function

Conventional kinesin is a tetrameric molecule composed of two heavy chains and two light chains, and transports various cargos along microtubules toward their plus ends. The heavy chains provide the motor activity, while the light chains bind to various cargos. This gene encodes a member of the kinesin light chain family. It associates with kinesin heavy chain through an N-terminal domain, and six tetratricopeptide repeat (TPR) motifs are thought to be involved in binding of cargos such as vesicles, mitochondria, and the Golgi complex. Thus, kinesin light chains function as adapter molecules and not motors per se. Although previously named ""kinesin 2"", this gene is not a member of the kinesin-2 / kinesin heavy chain subfamily of kinesin motor proteins. Extensive alternative splicing produces isoforms with different C-termini that are proposed to bind to different cargos; however, the full-length nature and/or biological validity of most of these variants have not been determined.

Background References

1. Moamer A. et. al. A role for kinesin-1 subunits KIF5B/KLC1 in regulating epithelial mesenchymal plasticity in breast tumorigenesis. EBioMedicine. 2019 Jul

2. Nguyen TQ. et. al. Characterization of the binding mode of JNK-interacting protein 1 (JIP1) to kinesin-light chain 1 (KLC1). J Biol Chem. 2018 Sep

Sequence Similarity

Belongs to the kinesin light chain family.

Tissue Specificity

Found in a variety of tissues. Mostly abundant in brain and spine.

Subcellular Location

Cytoskeleton, growth cone, cytoplasmic vesicle.

UNIPROT

SwissProt: Q07866 Human

SwissProt: O88447 Mouse

SwissProt: P37285 Rat

Synonyms

Kinesin 2 60/70kDa antibody

Kinesin light chain 1 antibody

Kinesin2 antibody

KLC 1 antibody

KLC antibody

Klc1 antibody

KLC1_HUMAN antibody

KNS 2 antibody

KNS 2A antibody

KNS2 antibody

Expand

Images

  • Western blot analysis of KLC1 on different lysates with Rabbit anti-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/500 dilution.<br /><br />Lane 1: Human brain tissue lysate (20 µg/Lane)<br />Lane 2: Rat brain tissue lysate (20 µg/Lane)<br />Lane 3: Hela cell lysate (10 µg/Lane)<br />Lane 4: PC-12 cell lysate (10 µg/Lane)<br /><br />Predicted band size: 65 kDa<br />Observed band size: 65 kDa<br /><br />Exposure time: 2 minutes;<br />10% 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/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/500 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 1/5,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of KLC1 on different lysates with Rabbit anti-KLC1 antibody (ET7110-72) at 1/500 dilution.

    Lane 1: Human brain tissue lysate (20 µg/Lane)
    Lane 2: Rat brain tissue lysate (20 µg/Lane)
    Lane 3: Hela cell lysate (10 µg/Lane)
    Lane 4: PC-12 cell lysate (10 µg/Lane)

    Predicted band size: 65 kDa
    Observed band size: 65 kDa

    Exposure time: 2 minutes;
    10% 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 (ET7110-72) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/5,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of MCF-7 cells labeling KLC1 with Rabbit anti-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of MCF-7 cells labeling KLC1 with Rabbit anti-KLC1 antibody (ET7110-72) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KLC1 antibody (ET7110-72) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

  • Immunocytochemistry analysis of RWPE-1 cells labeling KLC1 with Rabbit anti-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of RWPE-1 cells labeling KLC1 with Rabbit anti-KLC1 antibody (ET7110-72) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-KLC1 antibody (ET7110-72) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/50 dilution for 0.5 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 testis tissue with Rabbit anti-KLC1 antibody (ET7110-72) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET7110-72) at 1/50 dilution for 0.5 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 kidney tissue with Rabbit anti-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/200 dilution for 0.5 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 kidney tissue with Rabbit anti-KLC1 antibody (ET7110-72) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET7110-72) at 1/200 dilution for 0.5 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-KLC1 antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/400 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>) at 1/400 dilution for 0.5 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-KLC1 antibody (ET7110-72) at 1/400 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET7110-72) at 1/400 dilution for 0.5 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 KLC1 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7110-72" style="font-weight: bold;text-decoration: underline;">ET7110-72</a>, 1/50) (red). 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; black).

    Flow cytometric analysis of KLC1 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-72, 1/50) (red). 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; black).

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