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PDIA6 Recombinant Rabbit Monoclonal Antibody [JF0974]

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Specification

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Catalog# ET1702-26

PDIA6 Recombinant Rabbit Monoclonal Antibody [JF0974]

Application

  • WB

  • FC

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

PDIA6 Recombinant Rabbit Monoclonal Antibody [JF0974]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within PDIA6 aa 1-108 / 440.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, FC, IF-Cell, IHC-P

Molecular Weight

Predicted band size: 48 kDa

Positive Control

HepG2 cell lysate, K-562 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, 4T1 cell lysate, PC-12 cell lysate, Rat liver tissue lysate, PC-12, human liver tissue, mouse liver tissue, rat liver tissue.

Conjugation

unconjugated

Clone Number

JF0974

RRID

AB_3070289

Product Features

Form

Liquid

Concentration

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*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • FC

  • 1:1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000-1:5,000

Target

Function

Endoplasmic reticulum proteins (ERps) are widely expressed proteins that localize to the ER and may act as proteases, protein disulfide isomerases, thiol-disulfide oxidases or phospholipases. ERp5, also known as PDIA6 (protein disulfide isomerase family A, member 6) or TXNDC7 is a 440 amino acid protein that contains two thioredoxin domains and belongs to the protein disulfide isomerase family. Localized to the melanosome, as well as to the lumen of the endoplasmic reticulum, ERp5 functions to catalyze the rearrangement of disulfide bonds in a variety of different proteins. Via its catalytic activity, ERp5 is able to reduce the disulfide bond that binds MICA to tumor cells, thereby releasing MICA and reducing the rate of tumor expansion. Multiple isoforms of ERp5 exist due to alternative splicing events.

Background References

1. Groenendyk J et al. Interplay between the oxidoreductase PDIA6 and microRNA-322 controls the response to disrupted endoplasmic reticulum calcium homeostasis. Sci Signal 7:ra54 (2014).

2. Wang X et al. An six-amino acid motif in the alpha3 domain of MICA is the cancer therapeutic target to inhibit shedding. Biochem Biophys Res Commun 387:476-81 (2009).

Sequence Similarity

Belongs to the protein disulfide isomerase family.

Tissue Specificity

Expressed in platelets (at protein level).

Subcellular Location

Endoplasmic reticulum lumen, Cell membrane, Melanosome.

UNIPROT

SwissProt: Q15084 Human

SwissProt: Q922R8 Mouse

SwissProt: Q63081 Rat

Synonyms

Endoplasmic reticulum protein 5 antibody

ER protein 5 antibody

ERp5 antibody

P5 antibody

Pdia6 antibody

PDIA6_HUMAN antibody

Protein disulfide isomerase A6 antibody

Protein disulfide isomerase associated 6 antibody

Protein disulfide isomerase family A member 6 antibody

Protein disulfide isomerase P5 antibody

Expand

Images

  • Western blot analysis of PDIA6 on different lysates with Rabbit anti-PDIA6 antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</a>) at 1/2,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: K-562 cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: 4T1 cell lysate<br />Lane 6: PC-12 cell lysate<br />Lane 7: Rat liver tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 48 kDa<br />Observed band size: 48 kDa<br /><br />Exposure time: Lane 1-3: 25 seconds; Lane 4-7: 3 minutes; 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/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</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 PDIA6 on different lysates with Rabbit anti-PDIA6 antibody (ET1702-26) at 1/2,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: K-562 cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: 4T1 cell lysate
    Lane 6: PC-12 cell lysate
    Lane 7: Rat liver tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 48 kDa
    Observed band size: 48 kDa

    Exposure time: Lane 1-3: 25 seconds; Lane 4-7: 3 minutes; 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 (ET1702-26) 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 PDIA6 on different lysates with Rabbit anti-PDIA6 antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-PDIA6 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 48 kDa<br />Observed band size: 48 kDa<br /><br />Exposure time: 21 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/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</a>) at 1/2,000 dilution was used in primary antibody dilution (K1803) 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 PDIA6 on different lysates with Rabbit anti-PDIA6 antibody (ET1702-26) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-PDIA6 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 48 kDa
    Observed band size: 48 kDa

    Exposure time: 21 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 (ET1702-26) at 1/2,000 dilution was used in primary antibody dilution (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.

  • Immunocytochemistry analysis of PC-12 cells labeling PDIA6 with Rabbit anti-PDIA6 antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</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-PDIA6 antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</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 PDIA6 with Rabbit anti-PDIA6 antibody (ET1702-26) 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-PDIA6 antibody (ET1702-26) 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-PDIA6 antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</a>) at 1/5,000 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/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</a>) at 1/5,000 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 liver tissue with Rabbit anti-PDIA6 antibody (ET1702-26) at 1/5,000 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 (ET1702-26) at 1/5,000 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-PDIA6 antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</a>) at 1/5,000 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/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</a>) at 1/5,000 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-PDIA6 antibody (ET1702-26) at 1/5,000 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 (ET1702-26) at 1/5,000 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-PDIA6 antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</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/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</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-PDIA6 antibody (ET1702-26) 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 (ET1702-26) 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 PC-12 cells labeling PDIA6.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1702-26" style="font-weight: bold;text-decoration: underline;">ET1702-26</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 PDIA6.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-26, 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).

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Citation