ATF4 Recombinant antibody

ATF4 Uni-rAb^TM Recombinant Antibody for WB, IHC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IP, ELISA

Conjugate

Unconjugated

CloneNo.

5N3

Cat no : 81798-1-RR

Synonyms

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

---

Validation Data Gallery

• 

  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 1480 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).

• 

  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.

---

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; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0

Recommended 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

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.

Product Information

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

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: ATF4 fusion protein Ag1279
• 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
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• 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.