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DCTN1 / p150-glued Recombinant Rabbit Monoclonal Antibody [PSH13-45] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751464

DCTN1 / p150-glued Recombinant Rabbit Monoclonal Antibody [PSH13-45] - BSA and Azide free

Application

  • WB

  • IHC-Fr

  • IHC-P

  • IF-Cell

  • FC

  • IF-Tissue

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

DCTN1 / p150-glued Recombinant Rabbit Monoclonal Antibody [PSH13-45] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human DCTN1 aa 1,051-1,278.

Species Reactivity

Human, Mouse, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 142 kDa

Positive Control

HeLa cell lysate, HEK-293 cell lysate, SH-SY5Y cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, human brain tissue, mouse brain tissue, HEK-293, Neuro-2a, PC-12.

Conjugation

unconjugated

Clone Number

PSH13-45

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:15,000-1:50,000

  • IHC-Fr

  • 1:500

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

  • IF-Tissue

  • 1:200

  • IP

  • 1-2μg/sample

Target

Function

Dynactin subunit 1 is a protein that in humans is encoded by the DCTN1 gene. This gene encodes the largest subunit of dynactin, a macromolecular complex consisting of 23 subunits (11 individual proteins ranging in size from 22 to 150 kD). Dynactin binds to cytoplasmic dynein, dynein cargo adaptors, and microtubules. It is involved in a diverse array of cellular functions, including ER-to-Golgi transport, the centripetal movement of lysosomes and endosomes, spindle formation, chromosome movement, nuclear positioning, and axonogenesis. This subunit is commonly referred to p150-glued. It is present in two copies per dynactin complex and forms an ≈75 nm long flexible arm that extends from the main body of dynactin. The p150-glued arm contains binding sites for microtubules, the microtubule plus tip binding protein EB1, and the N-terminus of the dynein intermediate chain. Alternative splicing of this gene results in at least 2 functionally distinct isoforms: a ubiquitously expressed one and a brain-specific one. Based on its cytogenetic location, this gene is considered as a candidate gene for limb-girdle muscular dystrophy.

Background References

1. Vidrine DW et al. DCTN1-ALK gene fusion in inflammatory myofibroblastic tumor (IMT) of the CNS. Childs Nerv Syst. 2021 Jul

2. Deshimaru M et al. DCTN1 Binds to TDP-43 and Regulates TDP-43 Aggregation. Int J Mol Sci. 2021 Apr

Subcellular Location

Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, centriole, spindle, Nucleus envelope, cell cortex.

UNIPROT

SwissProt: Q14203 Human

SwissProt: O08788 Mouse

SwissProt: P28023 Rat

Synonyms

150 kDa dynein associated polypeptide antibody

150 kDa dynein-associated polypeptide antibody

DAP 150 antibody

DAP-150 antibody

DAP150 antibody

DCTN 1 antibody

DCTN1 antibody

DCTN1_HUMAN antibody

DP 150 antibody

DP-150 antibody

Expand

Images

  • Western blot analysis of DCTN1 / p150-glued on different lysates with Rabbit anti-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>) at 1/15,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HEK-293 cell lysate<br />Lane 3: SH-SY5Y cell lysate<br />Lane 4: Neuro-2a cell lysate<br />Lane 5: NIH/3T3 cell lysate<br />Lane 6: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 142 kDa<br />Observed band size: 150 kDa<br /><br />Exposure time: 8 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/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>) at 1/15,000 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 DCTN1 / p150-glued on different lysates with Rabbit anti-DCTN1 / p150-glued antibody (HA751464) at 1/15,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HEK-293 cell lysate
    Lane 3: SH-SY5Y cell lysate
    Lane 4: Neuro-2a cell lysate
    Lane 5: NIH/3T3 cell lysate
    Lane 6: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 142 kDa
    Observed band size: 150 kDa

    Exposure time: 8 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 (HA751464) at 1/15,000 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.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: brain<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: brain

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Not required

  • Application: IHC-Fr<br /><br />Species: Rat<br /><br />Site: brain<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Rat

    Site: brain

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Not required

  • Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</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/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</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 brain tissue with Rabbit anti-DCTN1 / p150-glued antibody (HA751464) 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 (HA751464) 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 brain tissue with Rabbit anti-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</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/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</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 brain tissue with Rabbit anti-DCTN1 / p150-glued antibody (HA751464) 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 (HA751464) 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 HEK-293 cells labeling DCTN1 / p150-glued with Rabbit anti-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 HEK-293 cells labeling DCTN1 / p150-glued with Rabbit anti-DCTN1 / p150-glued antibody (HA751464) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-DCTN1 / p150-glued antibody (HA751464) 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 (HA601187, 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.

  • Immunocytochemistry analysis of Neuro-2a cells labeling DCTN1 / p150-glued with Rabbit anti-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 Neuro-2a cells labeling DCTN1 / p150-glued with Rabbit anti-DCTN1 / p150-glued antibody (HA751464) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-DCTN1 / p150-glued antibody (HA751464) 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 (HA601187, 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.

  • Immunocytochemistry analysis of PC-12 cells labeling DCTN1 / p150-glued with Rabbit anti-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-DCTN1 / p150-glued antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</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/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</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 PC-12 cells labeling DCTN1 / p150-glued with Rabbit anti-DCTN1 / p150-glued antibody (HA751464) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-DCTN1 / p150-glued antibody (HA751464) 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 (HA601187, 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.

  • Flow cytometric analysis of HEK-293 cells labeling DCTN1 / p150-glued.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HEK-293 cells labeling DCTN1 / p150-glued.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751464, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Flow cytometric analysis of Neuro-2a cells labeling DCTN1 / p150-glued.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Neuro-2a cells labeling DCTN1 / p150-glued.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751464, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Flow cytometric analysis of PC-12 cells labeling DCTN1 / p150-glued.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of PC-12 cells labeling DCTN1 / p150-glued.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA751464, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • DCTN1 / p150-glued was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a> at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody 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/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751464" style="font-weight: bold;text-decoration: underline;">HA751464</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 50 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    DCTN1 / p150-glued was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751464 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751464 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: HA751464 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA751464 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute 50 seconds; ECL: K1801

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