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Calnexin Recombinant Rabbit Monoclonal Antibody [SN20-54]

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Specification

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Catalog# ET1611-86

Calnexin Recombinant Rabbit Monoclonal Antibody [SN20-54]

Application

  • WB

  • IP

  • IHC-P

  • IF-Tissue

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

Calnexin Recombinant Rabbit Monoclonal Antibody [SN20-54]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Calnexin aa 543-592 / 592.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IP, IHC-P, IF-Tissue

Molecular Weight

Predicted band size: 68 kDa

Positive Control

HeLa cell lysate, HepG2 cell lysate, MCF7 cell lysate, PANC-1 cell lysate, HAP1 cell lysate, human kidney tissue, human pancreas tissue, mouse kidney tissue, mouse liver tissue, rat kidney tissue, rat liver tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

SN20-54

RRID

AB_3070066

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

  • 1:2,000-1:10,000

  • IP

  • 1-2μg/sample

  • IHC-P

  • 1:1,000-1:10,000

  • IF-Tissue

  • 1:1,000

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. Noy PJ et al. TspanC8 Tetraspanins and A Disintegrin and Metalloprotease 10 (ADAM10) Interact via Their Extracellular Regions: EVIDENCE FOR DISTINCT BINDING MECHANISMS FOR DIFFERENT TspanC8 PROTEINS. J Biol Chem 291:3145-57 (2016).

2. Askautrud HA et al. Global gene expression analysis reveals a link between NDRG1 and vesicle transport. PLoS One 9:e87268 (2014).

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

SwissProt: P35565 Rat

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

  • Western blot analysis of Calnexin on different lysates with Rabbit anti-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: MCF7 cell lysate<br />Lane 4: PANC-1 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 68 kDa<br />Observed band size: 100 kDa<br /><br />Exposure time: 2 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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/2,000 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 Calnexin on different lysates with Rabbit anti-Calnexin antibody (ET1611-86) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: MCF7 cell lysate
    Lane 4: PANC-1 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 68 kDa
    Observed band size: 100 kDa

    Exposure time: 2 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 (ET1611-86) at 1/2,000 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.

  • <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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/1,000 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: 180 seconds; ECL: K1802;<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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/1,000 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.

    ☑ Knockdown (KD)

    Western blot analysis of Calnexin on different lysates with Rabbit anti-Calnexin antibody (ET1611-86) at 1/1,000 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: 180 seconds; ECL: K1802;

    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 (ET1611-86) at 1/1,000 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.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/10,000 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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/10,000 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-Calnexin antibody (ET1611-86) at 1/10,000 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 (ET1611-86) at 1/10,000 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 pancreas tissue with Rabbit anti-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/2,000 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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a>) at 1/2,000 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 pancreas tissue with Rabbit anti-Calnexin antibody (ET1611-86) at 1/2,000 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 (ET1611-86) at 1/2,000 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-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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 kidney tissue with Rabbit anti-Calnexin antibody (ET1611-86) 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 (ET1611-86) 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 liver tissue with Rabbit anti-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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 liver tissue with Rabbit anti-Calnexin antibody (ET1611-86) 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 (ET1611-86) 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-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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-Calnexin antibody (ET1611-86) 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 (ET1611-86) 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 liver tissue with Rabbit anti-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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 liver tissue with Rabbit anti-Calnexin antibody (ET1611-86) 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 (ET1611-86) 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 brain tissue with Rabbit anti-Calnexin antibody (<a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</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 brain tissue with Rabbit anti-Calnexin antibody (ET1611-86) 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 (ET1611-86) 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.

  • Calnexin was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) 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/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1611-86" style="font-weight: bold;text-decoration: underline;">ET1611-86</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 5 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 ET1611-86 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1611-86 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa cell lysate (input)
    Lane 2: ET1611-86 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of ET1611-86 in HeLa cell lysate

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

  • Application: IF-Tissue<br /><br />Species: Human<br /><br />Site: kidney<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/1,000

    Application: IF-Tissue

    Species: Human

    Site: kidney

    Sample: Paraffin-embedded section

    Antibody concentration: 1/1,000

  • Application: IF-Tissue<br /><br />Species: Mouse<br /><br />Site: kidney<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/1,000

    Application: IF-Tissue

    Species: Mouse

    Site: kidney

    Sample: Paraffin-embedded section

    Antibody concentration: 1/1,000

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Citation