Filter: AllWBIPIHCChIP-qPCRAffinity

WB analysis of HEK-293 using 81798-1-RR
Non-treated HEK-293 and MG132 treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 81798-1-RR (ATF4 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
WB analysis of A431 using 81798-1-RR
A431 cells were subjected to SDS PAGE followed by western blot with 81798-1-RR (ATF4 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.
WB analysis of HeLa using 81798-1-RR
Non-treated HeLa and MG132 treated HeLa cells were subjected to SDS PAGE followed by western blot with 81798-1-RR (ATF4 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.
IP experiment of HEK-293 using 81798-1-RR
IP result of anti-ATF4 (IP:81798-1-RR, 4ug; Detection:81798-1-RR 1:4000) with HEK-293 cells lysate 1440 ug.
IHC staining of human breast cancer using 81798-1-RR
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human breast cancer using 81798-1-RR
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human stomach cancer using 81798-1-RR
Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human stomach cancer using 81798-1-RR
Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human colon cancer using 81798-1-RR
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 81798-1-RR (ATF4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
ChIP experiment of NIH/3T3 using 81798-1-RR
Chromatin was prepared from NIH/3T3 cells treated with Tunicamycin (2ug/mll) overnight. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 20 µg of cross-linked chromatin, 5 µg of ATF4 (81798-1-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.
Affinity and Kinetic Characterization of 81798-1-RR
Biolayer interferometry (BLl) kinetic assays of 81798-1-RR against Human ATF4 were performed. The affinity constant is 0.500 nM.

ATF4 Recombinant monoclonal antibody

Name: ATF4 Recombinant monoclonal antibody
Brand: Proteintech
SKU: 81798-1-RR

ATF4 Uni-rAb^® Recombinant Antibody for WB, IHC, IP, ELISA, ChIP-qPCR

Cat No. 81798-1-RR

Clone No.5N3

Publications(3)

Review Product

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IP, ELISA, ChIP-qPCR

Synonyms

5N3, cAMP-dependent transcription factor ATF-4, CREB2, Cyclic AMP-responsive element-binding protein 2, Tax-responsive enhancer element-binding protein 67

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

PBS Only

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

Freight/Packing: -

Quantity

- +

Request bulk or custom quote

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Overview

Tested Applications
Product Information
Publications (3)
Protocols

Print datasheet

MSDS SDS

Tested Applications

Positive WB detected in	A431 cells, MG132 treated HEK-293 cells, MG132 treated HeLa cells
Positive IP detected in	HEK-293 cells
Positive IHC detected in	human breast cancer tissue, human stomach cancer tissue, human colon cancer tissue **Note: suggested antigen retrieval with TE buffer pH 9.0; () Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0***
Positive ChIP-qPCR detected in	Tunicamycin (2ug/mll) overnight NIH/3T3 cells

Recommended dilution

Application	Dilution
Western Blot (WB)	WB : 1:5000-1:50000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)	IHC : 1:50-1:500
CHIP-QPCR	CHIP-QPCR : 1:10-1:100
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Published Applications

WB	See 2 publications below

Product Information

81798-1-RR targets ATF4 in WB, IHC, IP, ELISA, ChIP-qPCR applications and shows reactivity with human, mouse samples.

Tested Reactivity	human, mouse
Cited Reactivity	human
Host / Isotype	Rabbit / IgG
Class	Recombinant
Type	Antibody
Immunogen	CatNo: Ag1279 Product name: Recombinant human ATF4 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-351 aa of BC022088 Sequence: MTEMSFLSSEVLVGDLMSPFDQSGLGAEESLGLLDDYLEVAKHFKPHGFSSDKAKAGSSEWLAVDGLVSPSNNSKEDAFSGTDWMLEKMDLKEFDLDALLGIDDLETMPDDLLTTLDDTCDLFAPLVQETNKQPPQTVNPIGHLPESLTKPDQVAPFTFLQPLPLSPGVLSSTPDHSFSLELGSEVDITEGDRKPDYTAYVAMIPQCIKEEDTPSDNDSGICMSPESYLGSPQHSPSTRGSPNRSLPSPGVLCGSARPKPYDPPGEKMVAAKVKGEKLDKKLKKMEQNKTAATRYRQKKRAEQEALTGECKELEKKNEALKERADSLAKEIQYLKDLIEEVRKARGKKRVP Predict reactive species
Full Name	activating transcription factor 4 (tax-responsive enhancer element B67)
Calculated Molecular Weight	39 kDa
Observed Molecular Weight	50 kDa
GenBank Accession Number	BC022088
Gene Symbol	ATF4
Gene ID (NCBI)	468
RRID	AB_2935580
Conjugate	Unconjugated
Form	Liquid
Purification Method	Protein A purification
UNIPROT ID	P18848
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

ATF4 is a transcription factor, that accumulates predominantly in osteoblasts, where it regulates terminal osteoblast differentiation and bone formation[PMID: 19016586]. As a basic leucine-zipper (bZip) transcription factor, ATF4 can regulate amino acid metabolism, cellular redox state, and anti-stress responses. It also regulates age-related and diet-induced obesity and glucose homeostasis in mammals, and has conserved metabolic functions in flies[PMID: 19726872]. Due to its location at chromosome 22q13, a region linked to schizophrenia, ATF4 is considered as a positional candidate gene for schizophrenia[PMID: 18163433]. Otherwise, since ATF4 is induced by tumour microenvironmental factors, and regulates processes relevant to cancer progression, it might serve as a potential therapeutic target in cancer. Endogenous ATF4 protein has a molecular mass of 50kd. [PMID: 17726049]. ATF4 can bind DNA as a homodimer and as a heterodimer. ATF4 is ubiquitinated by SCF(BTRC) in response to mTORC1 signal, followed by proteasomal degradation and leading to down-regulate expression of SIRT4, so the molecular weight of ATF4 may be 70 kDa.

Protocols

Product Specific Protocols
IHC protocol for ATF4 antibody 81798-1-RR	Download protocol
IP protocol for ATF4 antibody 81798-1-RR	Download protocol
WB protocol for ATF4 antibody 81798-1-RR	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
human	WB	Oncol Rep LRRC59 inhibits perk pathway‑induced apoptosis and promotes cell proliferation, migration and invasion in colorectal cancer cells. Authors - Xingdong Hou View Article
		Genes Dev Ribosome association inhibits stress-induced gene mRNA localization to stress granules Authors - Noah S Helton View Article
human	WB	Mol Med Rep Astragaloside IV promotes the apoptosis of pancreatic cancer cells by activating endoplasmic reticulum stress through the PERK/ATF4/CHOP signaling pathway. Authors - Yijun Wang View Article