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DDX39A Recombinant Rabbit Monoclonal Antibody [PSH05-57]

Usd: 385

Specification

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Catalog# HA722377

DDX39A Recombinant Rabbit Monoclonal Antibody [PSH05-57]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IP

  • IF-Tissue

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

DDX39A Recombinant Rabbit Monoclonal Antibody [PSH05-57]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human DDX39A aa 1-50 / 427.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IHC-P, IP, IF-Tissue

Molecular Weight

Predicted band size: 49 kDa

Positive Control

Jurkat cell lysate, HeLa cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, mouse testis tissue lysate, HeLa, human colon tissue, mouse colon tissue.

Conjugation

unconjugated

Clone Number

PSH05-57

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000-1:5,000

  • IP

  • 1-2μg/sample

  • IF-Tissue

  • 1:200-1:1,000

Target

Function

ATP-dependent RNA helicase DDX39 is an enzyme that in humans is encoded by the DDX39 gene. This gene encodes a member of the DEAD box protein family. These proteins are characterized by the conserved motif Asp-Glu-Ala-Asp (DEAD) and are putative RNA helicases. They are implicated in a number of cellular processes involving alteration of RNA secondary structure, such as translation initiation, nuclear and mitochondrial splicing, and ribosome and spliceosome assembly. Based on their distribution patterns, some members of the DEAD box protein family are believed to be involved in embryogenesis, spermatogenesis, and cellular growth and division.

Background References

1. Bao Y et al. DDX39 as a predictor of clinical prognosis and immune checkpoint therapy efficacy in patients with clear cell renal cell carcinoma. Int J Biol Sci. 2021 Jul

2. Xing C et al. DDX39 Overexpression Predicts a Poor Prognosis and Promotes Aggressiveness of Melanoma by Cooperating With SNAIL. Front Oncol. 2020 Aug

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: O00148 Human

SwissProt: Q8VDW0 Mouse

Synonyms

ATP dependent RNA helicase DDX39 antibody

ATP-dependent RNA helicase DDX39A antibody

BAT1 antibody

BAT1L antibody

DDX39 antibody

DDXL antibody

DEAD (Asp-Glu-Ala-Asp) box polypeptide 39 isoform 2 antibody

DEAD (Asp-Glu-Ala-Asp) box polypeptide 39A antibody

DEAD box protein 39 antibody

DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 39 antibody

Expand

Images

  • Western blot analysis of DDX39A on different lysates with Rabbit anti-DDX39A antibody (<a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: Jurkat cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: HepG2 cell lysate (20 µg/Lane)<br />Lane 4: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 5: Mouse testis tissue lysate (10 µg/Lane)<br /><br />Predicted band size: 49 kDa<br />Observed band size: 49 kDa<br /><br />Exposure time: 25 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/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were 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 DDX39A on different lysates with Rabbit anti-DDX39A antibody (HA722377) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: Jurkat cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: HepG2 cell lysate (20 µg/Lane)
    Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 5: Mouse testis tissue lysate (10 µg/Lane)

    Predicted band size: 49 kDa
    Observed band size: 49 kDa

    Exposure time: 25 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 (HA722377) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were 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 DDX39A on different lysates with Rabbit anti-DDX39A antibody (<a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a>) at 1/1,000 dilution.<br /><br />Lane 1: RPE-si NT cell lysate<br />Lane 2: RPE-si DDX39A cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 49 kDa<br />Observed band size: 49 kDa<br /><br />Exposure time: 42 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/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</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 DDX39A on different lysates with Rabbit anti-DDX39A antibody (HA722377) at 1/1,000 dilution.

    Lane 1: RPE-si NT cell lysate
    Lane 2: RPE-si DDX39A cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 49 kDa
    Observed band size: 49 kDa

    Exposure time: 42 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 (HA722377) 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.

  • Immunocytochemistry analysis of HeLa cells labeling DDX39A with Rabbit anti-DDX39A antibody (<a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-DDX39A antibody (<a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</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/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</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 HeLa cells labeling DDX39A with Rabbit anti-DDX39A antibody (HA722377) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-DDX39A antibody (HA722377) 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 (M1305-2, 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 colon tissue with Rabbit anti-DDX39A antibody (<a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</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 colon tissue with Rabbit anti-DDX39A antibody (HA722377) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA722377) 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 colon tissue with Rabbit anti-DDX39A antibody (<a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</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 colon tissue with Rabbit anti-DDX39A antibody (HA722377) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA722377) 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.

  • DDX39A was immunoprecipitated from 0.2 mg HeLa cell lysate with <a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</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/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a> IP in HeLa cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722377" style="font-weight: bold;text-decoration: underline;">HA722377</a> in HeLa cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 10 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    DDX39A was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722377 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722377 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: HA722377 IP in HeLa cell lysate
    Lane 3: Rabbit IgG instead of HA722377 in HeLa cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 10 seconds; ECL: K1801

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