ATF4 Rabbit pAb

Blank control:Mouse spleen.
Primary Antibody (green line): Rabbit Anti-ATF4 Alpha antibody (AP52146)
Dilution: 2 µg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1 µg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (ATF4) polyclonal Antibody, Unconjugated (AP52146) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded (mouse skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4) Polyclonal Antibody, Unconjugated (AP52146) at 1:100 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Sample:
Lane 1: Mouse Cerebrum tissue lysates
Lane 2: Rat Cerebrum tissue lysates
Lane 3: Human HeLa cell lysates
Lane 4: Human Jurkat cell lysates
Lane 5: Human HL-60 cell lysates
Primary: Anti-ATF4 (AP52146) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 38 kDa
Observed band size: 47 kDa
Paraformaldehyde-fixed, paraffin embedded (Human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4) Polyclonal Antibody, Unconjugated (AP52146) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

产品详情
实验流程
背景知识

Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, IHC-F, IF
Primary Accession	P18848
Reactivity	Human, Mouse, Rat
Predicted	Dog, Pig, Horse, Rabbit, Sheep
Host	Rabbit
Clonality	Polyclonal
Calculated MW	38 KDa
Physical State	Liquid
Immunogen	KLH conjugated synthetic peptide derived from human ATF4
Epitope Specificity	251-351/351
Isotype	IgG
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Cytoplasm. Cell membrane. Nucleus. Colocalizes with GABBR1 in hippocampal neuron dendritic membranes.
SIMILARITY	Belongs to the bZIP family. Contains 1 bZIP domain.
SUBUNIT	Binds DNA as a homo- or heterodimer. Interacts (via its leucine zipper domain) with GABBR1 and GABBR2 (via their C-termini). Interacts (via its DNA binding domain) with FOXO1 (C-terminal half); the interaction occurs in osteoblasts and regulates glucose homeostasis through suppression of beta-cell proliferation and a decrease in insulin production. Interacts with SATB2; the interaction results in enhanced DNA binding and transactivation by these transcription factors. Interacts with CEP290 (via an N-terminal region). Interacts with NEK6, DAPK2 (isoform 2) and ZIPK/DAPK3. Forms a heterodimer with TXLNG in osteoblasts. Interacts with DDIT3/CHOP.
Post-translational modifications	Ubiquitinated by SCF(BTRC) in response to mTORC1 signal, followed by proteasomal degradation and leading to down-regulate expression of SIRT4. Phosphorylated by NEK6. Phosphorylated on the betaTrCP degron motif at Ser-219, followed by phosphorylation at Thr-213, Ser-224, Ser-231, Ser-235 and Ser-248, promoting interaction with BTRC and ubiquitination. Phosphorylation is promoted by mTORC1. Phosphorylated by NEK6.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	ATF4 is a transcription factor that was originally identified as a widely expressed mammalian DNA binding protein that could bind a tax-responsive enhancer element in the LTR of HTLV1. The encoded protein was also isolated and characterized as the cAMP-response element binding protein 2 (CREB2). The protein encoded by this gene belongs to a family of DNA-binding proteins that includes the AP1 family of transcription factors, cAMP-response element binding proteins (CREBs) and CREB-like proteins. These transcription factors share a leucine zipper region that is involved in protein-protein interactions, located C-terminal to a stretch of basic amino acids that functions as a DNA binding domain (referenced from Entrez gene).

Additional Information
Other Names	Cyclic AMP-dependent transcription factor ATF-4, cAMP-dependent transcription factor ATF-4, Activating transcription factor 4, Cyclic AMP-responsive element-binding protein 2, CREB-2, cAMP-responsive element-binding protein 2, Tax-responsive enhancer element-binding protein 67, TaxREB67, ATF4 {ECO:0000303\|PubMed:2516827, ECO:0000312\|HGNC:HGNC:786}
Dilution	WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,ICC/IF=1:100-500,IF=1:100-500,Flow-Cyt=0.2 µg /test
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Research Areas

For Research Use Only. Not For Use In Diagnostic Procedures.

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

ATF4 is a transcription factor that was originally identified as a widely expressed mammalian DNA binding protein that could bind a tax-responsive enhancer element in the LTR of HTLV1. The encoded protein was also isolated and characterized as the cAMP-response element binding protein 2 (CREB2). The protein encoded by this gene belongs to a family of DNA-binding proteins that includes the AP1 family of transcription factors, cAMP-response element binding proteins (CREBs) and CREB-like proteins. These transcription factors share a leucine zipper region that is involved in protein-protein interactions, located C-terminal to a stretch of basic amino acids that functions as a DNA binding domain (referenced from Entrez gene).

REFERENCES

Tsujimoto A.,et al.J. Virol. 65:1420-1426(1991).
Karpinski B.A.,et al.Proc. Natl. Acad. Sci. U.S.A. 89:4820-4824(1992).
Dunham I.,et al.Nature 402:489-495(1999).
Hai T.,et al.Genes Dev. 3:2083-2090(1989).
Hai T.,et al.Genes Dev. 4:682-682(1990).

终于等到您。ABCEPTA（百远生物）抗体产品。
点击下方“我要评价 ”按钮提交您的反馈信息，您的反馈和评价是我们最宝贵的财富之一,
我们将在1-3个工作日内处理您的反馈信息。

如有疑问，联系：0512-88856768 tech-china@abcepta.com.

我要评价

¥ 1,500.00

Cat# AP52146

Size:

50 µl

100 µl

Price:

¥ 1,500.00

¥ 2,600.00

Quantity:

reduce add

Availability: 3-5 days

询价

ATF4 Rabbit pAb

ATF4 Rabbit pAb

BACKGROUND

REFERENCES

相关产品

多肽合成服务

多克隆抗体制备服务

单克隆抗体制备服务

抗体测序服务

蛋白表达服务

药物发现定制服务