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SRP54 Recombinant Rabbit Monoclonal Antibody [JE43-25]

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Specification

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Catalog# ET7109-32

SRP54 Recombinant Rabbit Monoclonal Antibody [JE43-25]

Application

  • WB

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

SRP54 Recombinant Rabbit Monoclonal Antibody [JE43-25]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human SRP54 aa 441-490 / 504.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC, IP

Molecular Weight

Predicted band size: 56 kDa

Positive Control

MCF-7 cell lysate, HepG2 cell lysate, rat brain tissue, human breast carcinoma tissue, human kidney tissue, mouse brain tissue, HepG2.

Conjugation

unconjugated

Clone Number

JE43-25

RRID

AB_3070920

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

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • 1:10-1:50

Target

Function

SRP54, a component of the signal recognition particle (SRP), recognizes the signal peptide of secretory proteins and interacts with the signal recognition particle receptor to target the ribosome and the associated nascent chain to the endoplasmic reticulum. By functional analysis showed that SRP54 binds to SRP RNA via the M-domain, but only in the presence of RNA-bound SRP19, and that it associates with the signal peptide of nascent polypeptide chains. RNA interaction requires the presence of a loop in the C-terminal M-domain. Signal peptide recognition most likely involves methionine-rich loops. During signal peptide recognition, SRP54 is positioned at the exit site close to ribosomal proteins L23a (602326) and L35. When SRP54 contacts the signal recognition particle receptor (182180), SRP54 is rearranged such that it is no longer close to L23a. This repositioning may allow the translocon to dock with the ribosome, leading to insertion of the signal peptide into the translocation channel.

Background References

1. Sakashita E et al. Human RNPS1 and its associated factors: a versatile alternative pre-mRNA splicing regulator in vivo. Mol Cell Biol 24:1174-1187(2004).

2. Kuglstatter A et al. Induced structural changes of 7SL RNA during the assembly of human signal recognition particle. Nat Struct Biol 9:740-744(2002).

Sequence Similarity

Belongs to the GTP-binding SRP family. SRP54 subfamily.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P61011 Human

SwissProt: P14576 Mouse

SwissProt: Q6AYB5 Rat

Synonyms

Signal recognition particle 54 kDa protein antibody

Signal recognition particle 54kDa antibody

SRP 54 antibody

SRP54 antibody

SRP54_HUMAN antibody

Images

  • Western blot analysis of SRP54 on different lysates with Rabbit anti-SRP54 antibody (<a href="/products/ET7109-32" style="font-weight: bold;text-decoration: underline;">ET7109-32</a>) at 1/500 dilution.<br /><br />Lane 1: MCF-7 cell lysate<br />Lane 2: HepG2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 56 kDa<br />Observed band size: 56 kDa<br /><br />Exposure time: 2 minutes;<br /><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/ET7109-32" style="font-weight: bold;text-decoration: underline;">ET7109-32</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 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SRP54 on different lysates with Rabbit anti-SRP54 antibody (ET7109-32) at 1/500 dilution.

    Lane 1: MCF-7 cell lysate
    Lane 2: HepG2 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 56 kDa
    Observed band size: 56 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 (ET7109-32) 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 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-32" style="font-weight: bold;text-decoration: underline;">ET7109-32</a>, 1/50) for 30 minutes 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 brain tissue using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-32, 1/50) for 30 minutes 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 breast carcinoma tissue using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-32" style="font-weight: bold;text-decoration: underline;">ET7109-32</a>, 1/50) for 30 minutes 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 breast carcinoma tissue using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-32, 1/50) for 30 minutes 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 using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-32" style="font-weight: bold;text-decoration: underline;">ET7109-32</a>, 1/50) for 30 minutes 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 using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-32, 1/50) for 30 minutes 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 using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-32" style="font-weight: bold;text-decoration: underline;">ET7109-32</a>, 1/50) for 30 minutes 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 using anti-SRP54 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-32, 1/50) for 30 minutes 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 SRP54 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7109-32" style="font-weight: bold;text-decoration: underline;">ET7109-32</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of SRP54 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7109-32, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Citation