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CLIC4 Recombinant Rabbit Monoclonal Antibody [JE65-09]

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Specification

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Catalog# HA722058

CLIC4 Recombinant Rabbit Monoclonal Antibody [JE65-09]

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

CLIC4 Recombinant Rabbit Monoclonal Antibody [JE65-09]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human CLIC4 aa 1-50 / 253.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, FC

Target Molecular Weight

Predicted band size: 29 kDa

Positive Control

HeLa cell lysate, JAR cell lysate, HepG2 cell lysate, RAW264.7 cell lysate, Neuro-2a cell lysate, C6 cell lysate, human kidney tissue lysate, rat kidney tissue lysate, HeLa.

Conjugation

unconjugated

Clone Number

JE65-09

RRID

AB_3096838

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). 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

  • IF-Cell

  • 1:200

  • FC

  • 1:100

Target

Function

Chloride intracellular channel 4, also known as CLIC4,p644H1,HuH1, is a eukaryotic gene. Chloride channels are a diverse group of proteins that regulate fundamental cellular processes including stabilization of cell membrane potential, transepithelial transport, maintenance of intracellular pH, and regulation of cell volume. Chloride intracellular channel 4 (CLIC4) protein, encoded by the clic4 gene, is a member of the p64 family; the gene is expressed in many tissues. These channels are implicated in angiogenesis, pulmonary hypertension, cancer, and cardioprotection from ischemia-reperfusion injury. They exhibit an intracellular vesicular pattern in PANC-1 cells (pancreatic cancer cells). CLIC4 binds to dynamin I, α-tubulin, β-actin, creatine kinase and two 14-3-3 isoforms.

Background References

1. Wang B et al. CLIC4 abrogation promotes epithelial-mesenchymal transition in gastric cancer. Carcinogenesis. 2020 Jul

2. Kleinjan ML et al. CLIC4 Regulates Endothelial Barrier Control by Mediating PAR1 Signaling via RhoA. Arterioscler Thromb Vasc Biol. 2023 Aug

Subcellular Location

Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, Cytoplasmic vesicle membrane, Nucleus, Cell membrane, Mitochondrion, Cell junction.

UNIPROT

SwissProt: Q9Y696 Human

SwissProt: Q9QYB1 Mouse

SwissProt: Q9Z0W7 Rat

Synonyms

Chloride intracellular channel 4 antibody

Chloride intracellular channel 4 (mitochondrial) antibody

Chloride intracellular channel 4 like antibody

Chloride intracellular channel protein 4 antibody

Clic4 antibody

CLIC4_HUMAN antibody

CLIC4L antibody

DKFZP566G223 antibody

FLJ38640 antibody

H1 antibody

Expand

Images

  • Western blot analysis of CLIC4 on different lysates with Rabbit anti-CLIC4 antibody (<a href="/products/HA722058" style="font-weight: bold;text-decoration: underline;">HA722058</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: JAR cell lysate (20 µg/Lane)<br />Lane 3: HepG2 cell lysate (20 µg/Lane)<br />Lane 4: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 5: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 6: C6 cell lysate (20 µg/Lane)<br />Lane 7: Human kidney tissue lysate (40 µg/Lane)<br />Lane 8: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 29 kDa<br />Observed band size: 29 kDa<br /><br />Exposure time: 24 seconds;<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/HA722058" style="font-weight: bold;text-decoration: underline;">HA722058</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.

    Western blot analysis of CLIC4 on different lysates with Rabbit anti-CLIC4 antibody (HA722058) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: JAR cell lysate (20 µg/Lane)
    Lane 3: HepG2 cell lysate (20 µg/Lane)
    Lane 4: RAW264.7 cell lysate (20 µg/Lane)
    Lane 5: Neuro-2a cell lysate (20 µg/Lane)
    Lane 6: C6 cell lysate (20 µg/Lane)
    Lane 7: Human kidney tissue lysate (40 µg/Lane)
    Lane 8: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

    Exposure time: 24 seconds;

    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 (HA722058) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of CLIC4 on different lysates with Rabbit anti-CLIC4 antibody (<a href="/products/HA722058" style="font-weight: bold;text-decoration: underline;">HA722058</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-CLIC4 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 29 kDa<br />Observed band size: 29 kDa<br /><br />Exposure time: 40 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/HA722058" style="font-weight: bold;text-decoration: underline;">HA722058</a>) at 1/5,000 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 CLIC4 on different lysates with Rabbit anti-CLIC4 antibody (HA722058) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-CLIC4 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 29 kDa
    Observed band size: 29 kDa

    Exposure time: 40 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 (HA722058) at 1/5,000 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.

  • Immunocytochemistry analysis of HeLa cells labeling CLIC4 with Rabbit anti-CLIC4 antibody (<a href="/products/HA722058" style="font-weight: bold;text-decoration: underline;">HA722058</a>) at 1/200 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-CLIC4 antibody (<a href="/products/HA722058" style="font-weight: bold;text-decoration: underline;">HA722058</a>) at 1/200 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.

    Immunocytochemistry analysis of HeLa cells labeling CLIC4 with Rabbit anti-CLIC4 antibody (HA722058) at 1/200 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-CLIC4 antibody (HA722058) at 1/200 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.

  • Flow cytometric analysis of HeLa cells labeling CLIC4.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722058" style="font-weight: bold;text-decoration: underline;">HA722058</a>, 1/100) (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 HeLa cells labeling CLIC4.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722058, 1/100) (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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