Name: Anti-ATF-4 antibody [EPR18111]
Brand: RabMAb
SKU: ab184909
Rating: 5 (4 reviews)

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Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18111] to ATF-4
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
Reacts with: Human

APC Carrier Free FITC PE

Product name

Anti-ATF-4 antibody [EPR18111]
See all ATF-4 primary antibodies
Description

Rabbit monoclonal [EPR18111] to ATF-4
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Human
Does not react with: Mouse, Rat
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal liver and fetal kidney lysates; A431, HeLa, whole cell lysates; untransfected HEK-293 whole cell lysate (control) lysate; HEK-293 whole cell lysate transfected with ATF-4 with GFP-tag lysate; HepG2 treated with 5 ÂµM MG-132 (ab141003) and 3 Âµg/ml tunicamycin (ab120296) for 6 hours whole cell lysate. IHC-P: Human bladder cancer and stomach cancer tissues. ICC/IF: HeLa and A431 cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18111
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human ATF-4 protein (ab109946)
Related Buffer
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Related Products

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab184909 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/80.
WB	(2)	1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 38 kDa).
IHC-P	(1)	1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/1000.
IP		1/50.

Notes
Flow Cyt (Intra) 1/80.
WB 1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 38 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
IP 1/50.

Function

Transcriptional activator. Binds the cAMP response element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. It binds to a Tax-responsive enhancer element in the long terminal repeat of HTLV-I.
Sequence similarities

Belongs to the bZIP family.
Contains 1 bZIP domain.
Cellular localization

Cytoplasm. Cell membrane. Nucleus. Colocalizes with GABBR1 in hippocampal neuron dendritic membranes.
Target information above from: UniProt accession P18848 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 468 Human
- Omim: 604064 Human
- SwissProt: P18848 Human
- Unigene: 496487 Human
Alternative names
- Activating transcription factor 4 antibody
- ATF 4 antibody
- ATF4 antibody
- ATF4 protein antibody
- ATF4_HUMAN antibody
- cAMP-dependent transcription factor ATF-4 antibody
- cAMP-responsive element-binding protein 2 antibody
- CREB 2 antibody
- CREB-2 antibody
- CREB2 antibody
- Cyclic AMP dependent transcription factor ATF 4 antibody
- Cyclic AMP response element binding protein 2 antibody
- Cyclic AMP-dependent transcription factor ATF-4 antibody
- Cyclic AMP-responsive element-binding protein 2 antibody
- DNA binding protein TAXREB67 antibody
- DNA-binding protein TAXREB67 antibody
- Tax Responsive Enhancer Element B67 antibody
- Tax-responsive enhancer element-binding protein 67 antibody
- TaxREB67 antibody
- TXREB antibody
see all

Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)

All lanes : Anti-ATF-4 antibody [EPR18111] (ab184909) at 1/1000 dilution

Lane 1 : Human fetal liver lysate
Lane 2 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Human fetal kidney lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 38 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 3 minutes; Lane 2 and 3: 30 seconds; Lane 4: 3 minutes.
Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATF-4 with ab184909 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on HeLa cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab184909 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)This image was courtesy of an annoymous AbReview

Formalin-fixed paraffin-embedded human colon cancer tissue stained for ATF-4 using ab184909 at 1/80 dilution (45 minute incubation) in immunohistochemical analysis. A polyclonal Goat Horse Radish Peroxidase antibody was used as the secondary.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)

Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue labeling ATF-4 with ab184909 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on human stomach cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)

All lanes : Anti-ATF-4 antibody [EPR18111] (ab184909) at 1/5000 dilution

Lane 1 : Untransfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (control)
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with ATT-4 with GFP-tag lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 38 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-ATF-4 antibody [EPR18111] (ab184909)

All lanes : Anti-ATF-4 antibody [EPR18111] (ab184909) at 1/1000 dilution

Lane 1 : Untreated HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (control)
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) treated with 5 µM MG-132 (ab141003) and 3 µg/ml tunicamycin (ab120296) for 6 hours whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 38 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID: 22095285).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF-4 antibody [EPR18111] (ab184909)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling ATF-4 with ab184909 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on human bladder cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-ATF-4 antibody [EPR18111] (ab184909)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling ATF-4 with ab184909 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on A431 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab184909 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-ATF-4 antibody [EPR18111] (ab184909)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell linefrom cervix adenocarcinoma) cells labeling ATF-4with ab184909 at 1/80 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-ATF-4 antibody [EPR18111] (ab184909)

ATF-4 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab184909 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab184909 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HeLa whole cell lysate, 10µg (Input).

Lane 2: ab184909 IP in HeLa whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab184909 in HeLa whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.
Anti-ATF-4 antibody [EPR18111] (ab184909)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab184909? Please let us know so that we can cite the reference in this datasheet.

ab184909 has been referenced in 44 publications.

Arioka Y et al. Chromosome 22q11.2 deletion causes PERK-dependent vulnerability in dopaminergic neurons. EBioMedicine 63:103138 (2021). PubMed: 33341442
Li KK et al. U three protein 14a (UTP14A) promotes tumour proliferation and metastasis via the PERK/eIF2a/GRP78 signalling pathway in oesophageal squamous cell carcinoma. J Cancer 12:134-140 (2021). PubMed: 33391409
Oku Y et al. Homeostasis of the ER redox state subsequent to proteasome inhibition. Sci Rep 11:8655 (2021). PubMed: 33883613
Bao J et al. Pharmacological Disruption of Phosphorylated Eukaryotic Initiation Factor-2a/Activating Transcription Factor 4/Indian Hedgehog Protects Intervertebral Disc Degeneration via Reducing the Reactive Oxygen Species and Apoptosis of Nucleus Pulposus Cells. Front Cell Dev Biol 9:675486 (2021). PubMed: 34164397
Rong K et al. Alendronate Alleviated Femoral Head Necrosis and Upregulated BMP2/EIF2AK3/EIF2A/ATF4 Pathway in Liquid Nitrogen Treated Rats. Drug Des Devel Ther 15:1717-1724 (2021). PubMed: 33935494

View all Publications for this product

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1-4 of 4 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (HEK293)

Gel Running Conditions

Reduced Denaturing (4-12%)

Loading amount

50 µg

Specification

HEK293

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 25 2021

Application

ChIP

Sample

Human Cell lysate - whole cell (HEK293)

Negative control

No antibody

Specification

HEK293

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1.1% Formaldehyde

Positive control

Histone H3 antibody

Abcam user community

Verified customer

Submitted Jun 25 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Colon cancer)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA

Permeabilization

Specification

Colon cancer

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Jan 19 2018

Application

Western blot

Sample

Dog Cell lysate - whole cell (OSW cells)

Gel Running Conditions

Reduced Denaturing (10 %)

Loading amount

100 µg

Specification

OSW cells

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

MS. Liubov Lobanova

Verified customer

Submitted Jul 22 2016

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Buffer

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (44)

Customer reviews and Q&As

Western blot abreview for Anti-ATF-4 antibody [EPR18111]

ChIP abreview for Anti-ATF-4 antibody [EPR18111]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ATF-4 antibody [EPR18111]

Western blot abreview for Anti-ATF-4 antibody [EPR18111]