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ATG7 Recombinant Rabbit Monoclonal Antibody [SC06-30]

Usd: 385

Specification

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Catalog# ET1610-53

ATG7 Recombinant Rabbit Monoclonal Antibody [SC06-30]

Application

  • WB

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

ATG7 Recombinant Rabbit Monoclonal Antibody [SC06-30]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human ATG7 aa 654-703 / 703.

Species Reactivity

Human, Rat

Validated Applications

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

Molecular Weight

Predicted band size: 78 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, THP-1 cell lysate, human tonsil tissue, human spleen tissue, human kidney tissue, human liver tissue, rat kidney tissue lysate.

Conjugation

unconjugated

Clone Number

SC06-30

RRID

AB_3069938

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

  • 1:500-1:2,000

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:1,000

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

Target

Function

E1-like activating enzyme involved in the 2 ubiquitin-like systems required for cytoplasm to vacuole transport (Cvt) and autophagy. Activates ATG12 for its conjugation with ATG5 as well as the ATG8 family proteins for their conjugation with phosphatidylethanolamine. Both systems are needed for the ATG8 association to Cvt vesicles and autophagosomes membranes. Required for autophagic death induced by caspase-8 inhibition. Required for mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Modulates p53/TP53 activity to regulate cell cycle and survival during metabolic stress. Plays also a key role in the maintenance of axonal homeostasis, the prevention of axonal degeneration, the maintenance of hematopoietic stem cells, the formation of Paneth cell granules, as well as in adipose differentiation. Plays a role in regulating the liver clock and glucose metabolism by mediating the autophagic degradation of CRY1 (clock repressor) in a time-dependent manner.

Background References

1. He G et al. Gadd45b prevents autophagy and apoptosis against rat cerebral neuron oxygen-glucose deprivation/reperfusion injury. Apoptosis 21:390-403 (2016).

2. Nazarko TY et al. Peroxisomal Atg37 binds Atg30 or palmitoyl-CoA to regulate phagophore formation during pexophagy. J Cell Biol 204:541-57 (2014).

Sequence Similarity

Belongs to the ATG7 family.

Tissue Specificity

Widely expressed, especially in kidney, liver, lymph nodes and bone marrow.

Post-translational Modification

Acetylated by EP300.

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: O95352 Human

SwissProt: Q641Y5 Rat

Synonyms

1810013K23Rik antibody

Apg 7 antibody

APG7 autophagy 7 like antibody

APG7 autophagy 7-like (S. cerevisiae) antibody

APG7 like antibody

APG7, S. cerevisiae, homolog of antibody

APG7-like antibody

APG7L antibody

ATG 7 antibody

ATG12-activating enzyme E1 ATG7 antibody

Expand

Images

  • Western blot analysis of ATG7 on different lysates with Rabbit anti-ATG7 antibody (<a href="/products/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: THP-1 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 78 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 3 minutes;<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/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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 ATG7 on different lysates with Rabbit anti-ATG7 antibody (ET1610-53) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: THP-1 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 78 kDa
    Observed band size: 70 kDa

    Exposure time: 3 minutes;

    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 (ET1610-53) 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.

  • Western blot analysis of ATG7 on rat kidney tissue lysates with Rabbit anti-ATG7 antibody (<a href="/products/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 40 µg/Lane.<br /><br />Predicted band size: 78 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 3 minutes;<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/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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 ATG7 on rat kidney tissue lysates with Rabbit anti-ATG7 antibody (ET1610-53) at 1/1,000 dilution.

    Lysates/proteins at 40 µg/Lane.

    Predicted band size: 78 kDa
    Observed band size: 70 kDa

    Exposure time: 3 minutes;
    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 (ET1610-53) 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Atg7 antibody. 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 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/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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 tonsil tissue using anti-Atg7 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-53, 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-Atg7 antibody. 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 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/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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-Atg7 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-53, 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 kidney tissue using anti-Atg7 antibody. 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 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/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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 kidney tissue using anti-Atg7 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-53, 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 liver tissue with Rabbit anti-ATG7 antibody (<a href="/products/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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 human liver tissue with Rabbit anti-ATG7 antibody (ET1610-53) 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 (ET1610-53) 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 Atg7 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Atg7 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-53, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ATG7 on different lysates with Rabbit anti-ATG7 antibody (<a href="/products/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa-si NT cell lysate<br />Lane 2: HeLa-si ATG7 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 78 kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 10 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/ET1610-53" style="font-weight: bold;text-decoration: underline;">ET1610-53</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.

    ☑ Knockdown (KD)

    Western blot analysis of ATG7 on different lysates with Rabbit anti-ATG7 antibody (ET1610-53) at 1/2,000 dilution.

    Lane 1: HeLa-si NT cell lysate
    Lane 2: HeLa-si ATG7 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 78 kDa
    Observed band size: 70 kDa

    Exposure time: 10 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 (ET1610-53) 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

Products with the same target and pathway

metafield image

ATG7 Recombinant Rabbit Monoclonal Antibody [SC06-30] - BSA and Azide free

Application: WB,IF-Tissue,IHC-P,IP,FC

Reactivity: Human,Rat

Conjugate: unconjugated