ATG4D Antibody | Affinity Biosciences

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Western blot analysis of extracts from K562 cells, using ATG4D antibody.

Western blot analysis of extracts from K562 cells, using ATG4D Antibody.

Figure 3: Immunofluorescence staining of histological sections from the patients with the homozygous pathogenic variant c.

Product:	ATG4D Antibody
Catalog:	DF3977
Description:	Rabbit polyclonal antibody to ATG4D
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse
Prediction:	Pig, Horse, Dog
Mol.Wt.:	53 KD; 53kD(Calculated).
Uniprot:	Q86TL0
RRID:	AB_2836337

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Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

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MS Report

HPLC Report

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:1000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse

Prediction:

Pig(80%), Horse(90%), Dog(90%)

Clonality:

Polyclonal

Specificity:

ATG4D Antibody detects endogenous levels of total ATG4D.

RRID:

AB_2836337
Cite Format: Affinity Biosciences Cat# DF3977, RRID:AB_2836337.

Conjugate:

Unconjugated.

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Storage:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Alias:

Fold/Unfold

9830134P12Rik; APG4 autophagy 4 homolog D; APG4 D; APG4, S. cerevisiae, homolog of, D; APG4D; ATG4 autophagy related 4 homolog D (S. cerevisiae); Atg4d; ATG4D_HUMAN; Atg4dl; AUT like 4 cysteine endopeptidase; AUT-like 4 cysteine endopeptidase; AUTL4; Autophagin 4; Autophagin-4; autophagy 4, S. cerevisiae, homolog of, D; autophagy related 4D, cysteine peptidase; Autophagy related cysteine endopeptidase 4; Autophagy related protein 4 homolog D; Autophagy-related cysteine endopeptidase 4; Autophagy-related protein 4 homolog D; Cysteine protease ATG4D; Cysteine protease involved in autophagy; MGC31226;

Immunogens

Immunogen:

Uniprot:

Q86TL0(ATG4D_HUMAN) >>Visit HPA database.

Gene(ID):

ATG4D(84971)

Expression:

Q86TL0 ATG4D_HUMAN:

Mainly expressed in skeletal muscle and, to a lower extent, in testis.

Sequence:

Q86TL0 ATG4D_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MNSVSPAAAQYRSSSPEDARRRPEARRPRGPRGPDPNGLGPSGASGPALGSPGAGPSEPDEVDKFKAKFLTAWNNVKYGWVVKSRTSFSKISSIHLCGRRYRFEGEGDIQRFQRDFVSRLWLTYRRDFPPLPGGCLTSDCGWGCMLRSGQMMLAQGLLLHFLPRDWTWAEGMGLGPPELSGSASPSRYHGPARWMPPRWAQGAPELEQERRHRQIVSWFADHPRAPFGLHRLVELGQSSGKKAGDWYGPSLVAHILRKAVESCSDVTRLVVYVSQDCTVYKADVARLVARPDPTAEWKSVVILVPVRLGGETLNPVYVPCVKELLRCELCLGIMGGKPRHSLYFIGYQDDFLLYLDPHYCQPTVDVSQADFPLESFHCTSPRKMAFAKMDPSCTVGFYAGDRKEFETLCSELTRVLSSSSATERYPMFTLAEGHAQDHSLDDLCSQLAQPTLRLPRTGRLLRAKRPSSEDFVFL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Horse

Dog

Pig

Bovine

Sheep

Xenopus

Zebrafish

Chicken

Rabbit

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q86TL0 As Substrate

Q86TL0

Site	PTM Type	Source
K64	Ubiquitination	Uniprot
S93	Phosphorylation	Uniprot
Y101	Phosphorylation	Uniprot
T167	Phosphorylation	Uniprot
K241	Ubiquitination	Uniprot
K258	Ubiquitination	Uniprot
S264	Phosphorylation	Uniprot
K298	Ubiquitination	Uniprot
K322	Ubiquitination	Uniprot
K388	Ubiquitination	Uniprot
Y398	Phosphorylation	Uniprot
K403	Ubiquitination	Uniprot
K464	Ubiquitination	Uniprot
S467	Phosphorylation	Uniprot

Research Backgrounds

Q86TL0_ATG4D_HUMAN

Function:

Cysteine protease required for the cytoplasm to vacuole transport (Cvt) and autophagy. Cleaves the C-terminal amino acid of ATG8 family proteins MAP1LC3 and GABARAPL2, to reveal a C-terminal glycine. Exposure of the glycine at the C-terminus is essential for ATG8 proteins conjugation to phosphatidylethanolamine (PE) and insertion to membranes, which is necessary for autophagy. Has also an activity of delipidating enzyme for the PE-conjugated forms.

Plays a role as an autophagy regulator that links mitochondrial dysfunction with apoptosis. The mitochondrial import of ATG4D during cellular stress and differentiation may play important roles in the regulation of mitochondrial physiology, ROS, mitophagy and cell viability.

PTMs:

Cleaved by CASP3 during apoptosis which leads to increased activity. The cleavage by CASP3 reveals a cryptic mitochondrial targeting sequence immediately downstream of their canonical caspase cleavage sites which leads to mitochondrial import of the protein.

Subcellular Location:

Cytoplasm.

Cytoplasm. Mitochondrion matrix.
Note: Imported into mitochondrial matrix after cleavage by CASP3 during oxidative stress and cell death.

Tissue Specificity:

Mainly expressed in skeletal muscle and, to a lower extent, in testis.

Family&Domains:

Residues 64 to 103 constitute a cryptic mitochondrial transit peptide which is revealed after cleavage by caspase of residues 1 to 63 upon oxidative stress and cell death. It acts then as a functional transit peptide, and allows the import of the cleaved protein into the mitochondria (PubMed:22441018).

Belongs to the peptidase C54 family.

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - other. (View pathway)

· Cellular Processes > Transport and catabolism > Autophagy - animal. (View pathway)

References

1). Pathogenic variants of ATG4D in infertile men with non‐obstructive azoospermia identified using whole‐exome sequencing. Clinical Genetics (PubMed: 33988247) [IF=3.5]

Application: IF/ICC Species: Human Sample: testicular tissue

Figure 3: Immunofluorescence staining of histological sections from the patients with the homozygous pathogenic variant c.374G>T:p.Arg125Leu in ATG4D. (A) Immunofluorescence staining of ATG4D in testicular tissue sections from the control and patients with the ATG4D variants. (B) Immunofluorescence staining of LC3B in the testicular tissue sections. (C) TUNEL assay of testicular tissue sections from the control and patients with the ATG4D variants. The control sample was obtained from a patient with obstructive azoospermia who had normal spermatogenesis. Scale bar = 50 μm.

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ATG4D Antibody - #DF3977