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SEC62 Recombinant Rabbit Monoclonal Antibody [JE62-80]

Usd: 385

Specification

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Catalog# HA721534

SEC62 Recombinant Rabbit Monoclonal Antibody [JE62-80]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

SEC62 Recombinant Rabbit Monoclonal Antibody [JE62-80]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human SEC62 aa256-350.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC

Molecular Weight

Predicted band size: 46 kDa

Positive Control

HeLa cell lysate, A549 cell lysate, HepG2 cell lysate, K-562 cell lysate, A431 cell lysate, HEK-293 cell lysate, PANC-1 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, human kidney tissue, mouse kidney tissue, rat kidney tissue, HeLa.

Conjugation

unconjugated

Clone Number

JE62-80

RRID

AB_3072650

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

  • 1:1,000

  • IHC-P

  • 1:1,000-1:3,000

  • FC

  • 1:1,000

Target

Function

The Sec61 complex is the central component of the protein translocation apparatus of the endoplasmic reticulum (ER) membrane. The protein encoded by this gene and SEC63 protein are found to be associated with ribosome-free SEC61 complex. It is speculated that Sec61-Sec62-Sec63 may perform post-translational protein translocation into the ER. The Sec61-Sec62-Sec63 complex might also perform the backward transport of ER proteins that are subject to the ubiquitin-proteasome-dependent degradation pathway. The encoded protein is an integral membrane protein located in the rough ER. Mediates post-translational transport of precursor polypeptides across endoplasmic reticulum (ER). Proposed to act as a targeting receptor for small presecretory proteins containing short and apolar signal peptides. Targets and properly positions newly synthesized presecretory proteins into the SEC61 channel-forming translocon complex, triggering channel opening for polypeptide translocation to the ER lumen.

Background References

1. Lang S., Benedix J., Fedeles S.V., Schorr S., Schirra C., Schaeuble N., Jalal C., Greiner M., Hassdenteufel S. Different effects of Sec61alpha, Sec62 and Sec63 depletion on transport of polypeptides into the endoplasmic reticulum of mammalian cells. J. Cell Sci. 125:1958-1969 (2012).

2. Hassdenteufel S., Johnson N., Paton A.W., Paton J.C., High S., Zimmermann R. Chaperone-Mediated Sec61 Channel Gating during ER Import of Small Precursor Proteins Overcomes Sec61 Inhibitor-Reinforced Energy Barrier. Cell Rep. 23:1373-1386 (2018).

Subcellular Location

Endoplasmic reticulum membrane.

UNIPROT

SwissProt: Q99442 Human

SwissProt: Q8BU14 Mouse

Entrez Gene: 294912 Rat

Synonyms

Dtrp1 antibody

FLJ32803 antibody

hTP-1 antibody

HTP1 antibody

Membrane protein SEC62, S.cerevisiae, homolog of antibody

OTTHUMP00000213390 antibody

sec62 antibody

SEC62 homolog (S. cerevisiae) antibody

SEC62_HUMAN antibody

TLOC1 antibody

Expand

Images

  • Western blot analysis of SEC62 on different lysates with Rabbit anti-SEC62 antibody (<a href="/products/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: HepG2 cell lysate<br />Lane 4: K-562 cell lysate<br />Lane 5: A431 cell lysate<br />Lane 6: HEK-293 cell lysate<br />Lane 7: PANC-1 cell lysate<br />Lane 8: NIH/3T3 cell lysate<br />Lane 9: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 46 kDa<br />Observed band size: 50 kDa<br /><br />Exposure time: 4 minutes;<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/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</a>) at 1/1,000 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 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SEC62 on different lysates with Rabbit anti-SEC62 antibody (HA721534) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: A549 cell lysate
    Lane 3: HepG2 cell lysate
    Lane 4: K-562 cell lysate
    Lane 5: A431 cell lysate
    Lane 6: HEK-293 cell lysate
    Lane 7: PANC-1 cell lysate
    Lane 8: NIH/3T3 cell lysate
    Lane 9: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

    Exposure time: 4 minutes;

    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 (HA721534) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-SEC62 antibody (<a href="/products/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</a>) at 1/3,000 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/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</a>) at 1/3,000 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 kidney tissue with Rabbit anti-SEC62 antibody (HA721534) at 1/3,000 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 (HA721534) at 1/3,000 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 kidney tissue with Rabbit anti-SEC62 antibody (<a href="/products/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</a>) at 1/3,000 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/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</a>) at 1/3,000 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 kidney tissue with Rabbit anti-SEC62 antibody (HA721534) at 1/3,000 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 (HA721534) at 1/3,000 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 kidney tissue with Rabbit anti-SEC62 antibody (<a href="/products/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</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/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</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 kidney tissue with Rabbit anti-SEC62 antibody (HA721534) 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 (HA721534) 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.

  • Flow cytometric analysis of HeLa cells labeling SEC62.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721534" style="font-weight: bold;text-decoration: underline;">HA721534</a>, 1ug/ml) (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 HeLa cells labeling SEC62.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721534, 1ug/ml) (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).

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