ITCH Polyclonal antibody

• Featured Product
• KD/KO Validated

ITCH Polyclonal Antibody for WB, IP, IHC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (1)

Applications

WB, IHC, IP, CoIP, ELISA

Conjugate

Unconjugated

Publications

14

Reviews (1)

Cat no : 20920-1-AP

Synonyms

AIF4, AIP4, ITCH, NAPP1, NFE2 associated polypeptide 1

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Validation Data Gallery

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  WB analysis using 20920-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 20920-1-AP (ITCH antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  WB analysis of K-562 using 20920-1-AP

  K-562 cells were subjected to SDS PAGE followed by western blot with 20920-1-AP (ITCH antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of U-251 using 20920-1-AP

  U-251 cells were subjected to SDS PAGE followed by western blot with 20920-1-AP (ITCH antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of NIH/3T3 using 20920-1-AP

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 20920-1-AP (ITCH antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 20920-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 20920-1-AP (ITCH antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 20920-1-AP

  HeLa lysates prepared and IP of ITCH performed using 2.0 μg of 20920-1-AP coupled to protein A- Sepharose beads. The Ponceau stained transfers of each blot are shown. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

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  IP experiment of HEK-293 using 20920-1-AP

  IP result of anti-ITCH (IP:20920-1-AP, 4ug; Detection:20920-1-AP 1:300) with HEK-293 cells lysate 2400 ug.

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  IHC staining of human breast cancer using 20920-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 20920-1-AP (ITCH antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 20920-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 20920-1-AP (ITCH antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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Tested Applications

• Positive WB detected in: HEK-293 cells, U-251 cells, NIH/3T3 cells, K-562 cells, Hela cells, Raji cells
• Positive IP detected in: HEK-293 cells, HeLa cells
• Positive IHC detected in: human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 4 publications below
• WB: See 13 publications below
• IHC: See 1 publications below
• IP: See 2 publications below
• CoIP: See 3 publications below

Product Information

20920-1-AP targets ITCH in WB, IHC, IP, CoIP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: ITCH fusion protein Ag15120
• Full Name: itchy E3 ubiquitin protein ligase homolog (mouse)
• Calculated Molecular Weight: 903 aa, 103 kDa
• Observed Molecular Weight: 103 kDa, 70 kDa
• GenBank Accession Number: BC011571
• Gene Symbol: ITCH
• Gene ID (NCBI): 83737
• RRID: AB_2878765
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

ITCH is an essential component of a ubiquitin-editing protein complex, comprising also TNFAIP3, TAX1BP1 and RNF11, that ensures the transient nature of inflammatory signaling pathways. It is involved in the regulation of apoptosis and reactive oxygen species levels through the ubiquitination and proteasomal degradation of TXNIP. ITCH can be cleaved by caspase-3, -6, and -7 on residue Asp240 and form a 65 kDa cleaved form (PMID: 19541840).

Publications

Species	Application	Title
human	WB	Nat Cell Biol; Heat stress activates YAP/TAZ to induce the heat shock transcriptome.; Authors - Min Luo ; KD Validated
human	WB	Dev Cell; ASK1-Mediated Phosphorylation Blocks HDAC6 Ubiquitination and Degradation to Drive the Disassembly of Photoreceptor Connecting Cilia.; Authors - Jie Ran ; KD Validated
human	WB	Theranostics; ACSS3 represses prostate cancer progression through downregulating lipid droplet-associated protein PLIN3.; Authors - Lijie Zhou
human	WB,IP	Clin Transl Med; GRAMD4 inhibits tumour metastasis by recruiting the E3 ligase ITCH to target TAK1 for degradation in hepatocellular carcinoma.; Authors - Qian Yun Ge ; KD Validated
human	WB, CoIP	Front Oncol; Coiled-Coil Domain-Containing 68 Downregulation Promotes Colorectal Cancer Cell Growth by Inhibiting ITCH-Mediated CDK4 Degradation.; Authors - Cong Wang ; KD Validated
human	WB	J Mol Med (Berl); Hyper-SUMOylation of SMN induced by SENP2 deficiency decreases its stability and leads to spinal muscular atrophy-like pathology.; Authors - Yuhong Zhang

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Chrisovalantou (Verified Customer) (10-31-2024)

Antibody was able to detech endogenous ITCH in SH-SY5Y and HEK293T on western blot

• Applications: Western Blot
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: SH-SY5Y, HEK293T

ITCH Antibody Western Blot validation (1:1000 dilution) in SH-SY5Y, HEK293T (Cat no:20920-1-AP)