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Calnexin Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER1803-42

Calnexin Rabbit Polyclonal Antibody

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

Calnexin Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within human Calnexin aa 555-592/592.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC, IP

Target Molecular Weight

Predicted band size: 68 kDa

Positive Control

Daudi cell lysate, mouse lung tissue lysate, human kidney tissue, SKOV-3, Hela cell lysate, rat lung tissue lysate, rat brain tissue lysate, rat brain tissue, human brain tissue, mouse brain tissue.

Conjugation

unconjugated

RRID

AB_3069238

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P FC IP
human
Tested Tested
Tested Tested
Tested Tested
Tested Tested
mouse
Tested Tested
Tested Tested
 
 
rat
Tested Tested
Tested Tested
 
 

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:2,000- 1:10,000

  • IHC-P

  • 1:200-1:500

  • FC

  • 1:50-1:100

  • IP

  • 1-2μg/sample

Target

Function

Calnexin and Calregulin (also called calreticulin) are calcium-binding proteins that are localized to the endoplasmic reticulum, Calnexin to the membrane and Calregulin to the lumen. Calnexin is a type I membrane protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may play a role in assisting with protein assembly and in retaining unassembled protein subunits in the endoplasmic reticulum. Calregulin has both low- and high-affinity calcium-binding sites. Neither Calnexin nor Calregulin contains the calcium-binding “E-F hand” motif found in calmodulins. Calnexin and Calregulin are important for the maturation of glycoproteins in the endoplasmic reticulum and appear to bind many of the same proteins.

Background References

1. Lakkaraju A K et al. Palmitoylated calnexin is a key component of the ribosome-translocon complex. EMBO J 31:1823-1835 (2012).

2. Basrur V et al. Proteomic analysis of early melanosomes: identification of novel melanosomal proteins. J Proteome Res 2:69-79 (2003).

Sequence Similarity

Belongs to the calreticulin family.

Post-translational Modification

Phosphorylated at Ser-564 by MAPK3/ERK1. phosphorylation by MAPK3/ERK1 increases its association with ribosomes (By similarity).; Palmitoylation by DHHC6 leads to the preferential localization to the perinuclear rough ER. It mediates the association of calnexin with the ribosome-translocon complex (RTC) which is required for efficient folding of glycosylated proteins.; Ubiquitinated, leading to proteasomal degradation. Probably ubiquitinated by ZNRF4.

Subcellular Location

Endoplasmic reticulum membrane, Endoplasmic reticulum, Melanosome.

UNIPROT

SwissProt: P27824 Human

SwissProt: P35564 Mouse

Synonyms

Calnexin antibody

CALX_HUMAN antibody

CANX antibody

CNX antibody

FLJ26570 antibody

Histocompatibility complex class I antigen binding protein p88 antibody

IP90 antibody

Major histocompatibility complex class I antigen-binding protein p88 antibody

p90 antibody

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Calnexin on different lysates with Rabbit anti-Calnexin antibody (<a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/10,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Calnexin KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 68 kDa<br />Observed band size: 90 kDa<br /><br />Exposure time: 20 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/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/10,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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 Calnexin on different lysates with Rabbit anti-Calnexin antibody (ER1803-42) at 1/10,000 dilution.

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

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 68 kDa
    Observed band size: 90 kDa

    Exposure time: 20 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 (ER1803-42) at 1/10,000 dilution was used in 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.

  • Western blot analysis of Calnexin on different lysates with Rabbit anti-Calnexin antibody (<a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/2,000 dilution.<br /><br />Lane 1: Hela cell lysate（10 µg/Lane）<br />Lane 2: Rat lung tissue lysate（20 µg/Lane）<br />Lane 3: Rat brain tissue lysate（20 µg/Lane）<br /><br />Predicted band size: 68 kDa<br />Observed band size: 90 kDa<br /><br />Exposure time: 3 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/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/2,000 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Calnexin on different lysates with Rabbit anti-Calnexin antibody (ER1803-42) at 1/2,000 dilution.

    Lane 1: Hela cell lysate（10 µg/Lane）
    Lane 2: Rat lung tissue lysate（20 µg/Lane）
    Lane 3: Rat brain tissue lysate（20 µg/Lane）

    Predicted band size: 68 kDa
    Observed band size: 90 kDa

    Exposure time: 3 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 (ER1803-42) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 kidney tissue with Rabbit anti-Calnexin antibody (<a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/500 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/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/500 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-Calnexin antibody (ER1803-42) at 1/500 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 (ER1803-42) at 1/500 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-Calnexin antibody (<a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/200 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/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/200 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-Calnexin antibody (ER1803-42) at 1/200 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 (ER1803-42) at 1/200 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-Calnexin antibody (<a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/200 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/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/200 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-Calnexin antibody (ER1803-42) at 1/200 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 (ER1803-42) at 1/200 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 brain tissue with Rabbit anti-Calnexin antibody (<a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/200 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/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a>) at 1/200 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 brain tissue with Rabbit anti-Calnexin antibody (ER1803-42) at 1/200 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 (ER1803-42) at 1/200 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 SKOV-3 cells labeling Calnexin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</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 SKOV-3 cells labeling Calnexin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ER1803-42, 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).

  • Calnexin was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs&reg; VHH(HRP) (<a href="/products/025-100-005" style="font-weight: bold;text-decoration: underline;">025-100-005</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/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ER1803-42" style="font-weight: bold;text-decoration: underline;">ER1803-42</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 23 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Calnexin was immunoprecipitated from 0.2 mg HeLa cell lysate with ER1803-42 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ER1803-42 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: ER1803-42 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of ER1803-42 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 23 seconds; ECL: K1801

Citation

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

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Reactivity: Human,Mouse,Rat

Conjugate: unconjugated