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ULK1 Recombinant Rabbit Monoclonal Antibody [JA58-36]

Usd: 205

Specification

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Catalog# ET1704-63

ULK1 Recombinant Rabbit Monoclonal Antibody [JA58-36]

Application

  • WB

  • IHC-P

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

ULK1 Recombinant Rabbit Monoclonal Antibody [JA58-36]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human ULK1 aa 930-979 / 1050.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 113 kDa

Positive Control

A-172 cell lysate, SH-SY5Y cell lysate, U-2 OS cell lysate, MCF7 cell lysate, mouse spleen tissue lysate, rat spleen tissue lysate, HepG2 cell lysate, C2C12 cell lysate, MEF cell lysate, PC-12 cell lysate, human spleen tissue, rat skeletal muscle tissue, human colon carcinoma tissue, mouse brain tissue.

Conjugation

unconjugated

Clone Number

JA58-36

RRID

AB_3070509

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

  • 1:5,000

  • IHC-P

  • 1:50-1:200

Target

Function

Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3-kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2 and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. May also phosphorylate SESN2 and SQSTM1 to regulate autophagy.

Background References

1. Desantis, A. et al. Che-1-induced inhibition of mTOR pathway enables stress-induced autophagy. The EMBO journal. 34: 1214-30 (2015).

2. Xiong, H. et al. Activation of miR-34a/SIRT1/p53 signaling contributes to cochlear hair cell apoptosis: implications for age-related hearing loss. Neurobiology of aging. 36: 1692-701 (2015).

Sequence Similarity

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. APG1/unc-51/ULK1 subfamily.

Tissue Specificity

Ubiquitously expressed. Detected in the following adult tissues: skeletal muscle, heart, pancreas, brain, placenta, liver, kidney, and lung.

Post-translational Modification

Autophosphorylated. Phosphorylated under nutrient-rich conditions; dephosphorylated during starvation or following treatment with rapamycin. Under nutrient sufficiency, phosphorylated by MTOR/mTOR, disrupting the interaction with AMPK and preventing activation of ULK1 (By similarity). In response to nutrient limitation, phosphorylated and activated by AMPK, leading to activate autophagy.; Acetylated by KAT5/TIP60 under autophagy induction, promoting protein kinase activity.

Subcellular Location

Cytosol, Preautophagosomal structure.

UNIPROT

SwissProt: O75385 Human

SwissProt: O70405 Mouse

SwissProt: D3ZMG0 Rat

Synonyms

ATG 1 antibody

ATG1 antibody

ATG1 autophagy related 1 homolog antibody

ATG1A antibody

Autophagy related protein 1 homolog antibody

Autophagy-related protein 1 homolog antibody

FLJ38455 antibody

FLJ46475 antibody

hATG1 antibody

KIAA0722 antibody

Expand

Images

  • Western blot analysis of ULK1 on different lysates with Rabbit anti-ULK1 antibody (<a href="/products/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>) at 1/5,000 dilution.<br /><br />Lane 1: A-172 cell lysate (15 µg/Lane)<br />Lane 2: SH-SY5Y cell lysate (15 µg/Lane)<br />Lane 3: U-2 OS cell lysate (15 µg/Lane)<br />Lane 4: MCF7 cell lysate (15 µg/Lane)<br />Lane 5: Mouse spleen tissue lysate (20 µg/Lane)<br />Lane 6: Rat spleen tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 113 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 1 minute 30 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/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>) at 1/5,000 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 ULK1 on different lysates with Rabbit anti-ULK1 antibody (ET1704-63) at 1/5,000 dilution.

    Lane 1: A-172 cell lysate (15 µg/Lane)
    Lane 2: SH-SY5Y cell lysate (15 µg/Lane)
    Lane 3: U-2 OS cell lysate (15 µg/Lane)
    Lane 4: MCF7 cell lysate (15 µg/Lane)
    Lane 5: Mouse spleen tissue lysate (20 µg/Lane)
    Lane 6: Rat spleen tissue lysate (20 µg/Lane)

    Predicted band size: 113 kDa
    Observed band size: 130 kDa

    Exposure time: 1 minute 30 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 (ET1704-63) at 1/5,000 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.

  • Western blot analysis of ULK1 on different lysates with Rabbit anti-ULK1 antibody (<a href="/products/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>) at 1/5,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: C2C12 cell lysate<br />Lane 3: MEF cell lysate<br />Lane 4: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 113 kDa<br />Observed band size: 130 kDa<br /><br />Exposure time: 6 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/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>) at 1/5,000 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 ULK1 on different lysates with Rabbit anti-ULK1 antibody (ET1704-63) at 1/5,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: C2C12 cell lysate
    Lane 3: MEF cell lysate
    Lane 4: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 113 kDa
    Observed band size: 130 kDa

    Exposure time: 6 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 (ET1704-63) at 1/5,000 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 spleen tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>, 1/50) for 30 minutes 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 spleen tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-63, 1/50) for 30 minutes 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 skeletal muscle tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>, 1/50) for 30 minutes 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 skeletal muscle tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-63, 1/50) for 30 minutes 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 carcinoma tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>, 1/50) for 30 minutes 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 carcinoma tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-63, 1/50) for 30 minutes 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 brain tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-63" style="font-weight: bold;text-decoration: underline;">ET1704-63</a>, 1/50) for 30 minutes 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 brain tissue using anti-ULK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-63, 1/50) for 30 minutes 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.

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Citation

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