ATF5 Recombinant Rabbit Monoclonal Antibody [SD2099]
Overview
Product Name
ATF5 Recombinant Rabbit Monoclonal Antibody [SD2099]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human ATF5 aa 203-252 / 282.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Molecular Weight
Predicted band size: 31 kDa
Positive Control
HEK-293 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, HeLa, RAW264.7, C6, human liver tissue, human stomach tissue, mouse liver tissue, mouse stomach tissue, rat liver tissue, rat stomach tissue.
Conjugation
unconjugated
Clone Number
SD2099
RRID
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
-
1:2,000
-
IF-Cell
-
1:100
-
IF-Tissue
-
1:50-1:200
-
IHC-P
-
1:200-1:1,000
-
IP
-
1-2μg/sample
-
FC
-
1:1,000
Target
Function
Eukaryotic gene transcription is regulated by sequence-specific transcription factors that bind modular cis acting promoter and enhancer elements. The ATF/CREB transcription factor family binds the palindromic cAMP response element (CRE) octanucleotide TGACGTCA. The best characterized members of this gene family include CREB-1, CREB-2 (also designated ATF-4), CRE-BPa, LZIP (also designated CREB3 and Luman), CREM-1, CREM-2, ATF-1, ATF-2, ATF-3, ATF-5, ATF-6 and ATF-7. This family of proteins contain highly divergent N-terminal domains, but share a C-terminal leucine zipper for dimerization and DNA binding. ATF-5 (ATFx), which can localize to the cytoplasm or the nucleus, binds DNA as a dimer. It interacts with CCND3 and PTP4A1.
Background References
1. Liu X et al. Nucleophosmin (NPM1/B23) Interacts with Activating Transcription Factor 5 (ATF5) Protein and Promotes Proteasome- and Caspase-dependent ATF5 Degradation in Hepatocellular Carcinoma Cells. J Biol Chem 287:19599-609 (2012).
2. Dluzen D et al. BCL-2 is a downstream target of ATF5 that mediates the prosurvival function of ATF5 in a cell type-dependent manner. J Biol Chem 286:7705-13 (2011).
Sequence Similarity
Belongs to the bZIP family.
Tissue Specificity
Widely expressed with higher expression levels in liver.
Post-translational Modification
Ubiquitinated by CDC34 and UBE2B in order to be degraded by the proteasome. Cisplatin inhibits ubiquitination and proteasome-mediated degradation by inhibiting the interaction with CDC34. Ubiquitination and degradation by the proteasome are inhibited by NLK in a kinase-independent manner.; Phosphorylated by NLK, probably at Ser-92, Thr-94, Ser-126 and Ser-190.; Acetylated at Lys-29 by EP300, the acetylation enhances the interaction with CEBPB, DNA-binding and transactivation activity.
Subcellular Location
Centrosome, Nucleus, Cytoplasm.
Synonyms
Activating transcription factor 5 alpha/beta antibody
Activating transcription factor 5 antibody
Activating transcription factor X antibody
AFTA antibody
ATF 5 antibody
ATF 7 antibody
ATF7 antibody
ATFX antibody
BZIP protein ATF7 antibody
cAMP dependent transcription factor ATF 5 antibody
ExpandImages
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Western blot analysis of ATF5 on different lysates with Rabbit anti-ATF5 antibody (ET1612-38) at 1/2,000 dilution.
Lane 1: HEK-293 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: C6 cell lysate
Lane 6: PC-12 cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 31 kDa
Observed band size: 35 kDa
Exposure time: 28 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 (ET1612-38) 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. -
Immunocytochemistry analysis of HeLa cells labeling ATF5 with Rabbit anti-ATF5 antibody (ET1612-38) at 1/100 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-ATF5 antibody (ET1612-38) at 1/100 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.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of RAW264.7 cells labeling ATF5 with Rabbit anti-ATF5 antibody (ET1612-38) at 1/100 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-ATF5 antibody (ET1612-38) at 1/100 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.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling ATF5 with Rabbit anti-ATF5 antibody (ET1612-38) at 1/100 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-ATF5 antibody (ET1612-38) at 1/100 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.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of HeLa cells labeling ATF5.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-38, 1/1,000) (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). -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-ATF5 antibody (ET1612-38) at 1/200 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-38) at 1/200 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 stomach tissue with Rabbit anti-ATF5 antibody (ET1612-38) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-38) 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 mouse liver tissue with Rabbit anti-ATF5 antibody (ET1612-38) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-38) 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 mouse stomach tissue with Rabbit anti-ATF5 antibody (ET1612-38) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-38) 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 rat liver tissue with Rabbit anti-ATF5 antibody (ET1612-38) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-38) 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 rat stomach tissue with Rabbit anti-ATF5 antibody (ET1612-38) at 1/1,000 dilution.
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 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-38) 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. -
ATF5 was immunoprecipitated from 0.2 mg Jurkat cell lysate with ET1612-38 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1612-38 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: Jurkat cell lysate (input)
Lane 2: ET1612-38 IP in Jurkat cell lysate
Lane 3: Rabbit IgG instead of ET1612-38 in Jurkat cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 8 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
-
Agarononaose from red algae delays cardiomyocyte aging by improving mitochondrial function via enhanced oxidative phosphorylation: A mitochondrial transplantation perspective
Journal: Journal Of Functional Foods
DOI: 10.1016/j.jff.2025.107131
IF: 4
Application: WB
Reactivity: Mouse
Publish date: 2026 Jan