PRODUCT CODE: ET1612-37

Recombinant ATF4 Monoclonal Antibody (ET1612-37)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Rat

Western blot analysis of ATF4 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-37, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Hela cell lysate<br />
 Lane 2: PC-12 cell lysate<br />
 Lane 3: HL-60 cell lysate<br />
 Lane 4: K562 cell lysate<br />
 Lane 5: human lung carcinoma tissue lysate
  • Western blot analysis of ATF4 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-37, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Hela cell lysate<br />
 Lane 2: PC-12 cell lysate<br />
 Lane 3: HL-60 cell lysate<br />
 Lane 4: K562 cell lysate<br />
 Lane 5: human lung carcinoma tissue lysate
  • ICC staining of ATF4 in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1612-37, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-ATF4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37, 1/50) for 30 minutes 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 prostate carcinoma tissue using anti-ATF4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37, 1/50) for 30 minutes 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 skin tissue using anti-ATF4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37, 1/50) for 30 minutes 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 breast carcinoma tissue using anti-ATF4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37, 1/50) for 30 minutes 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 carcinoma tissue using anti-ATF4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37, 1/50) for 30 minutes 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 small intestine tissue using anti-ATF4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37, 1/50) for 30 minutes 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 colon carcinoma tissue using anti-ATF4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37, 1/50) for 30 minutes 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.
  • Flow cytometric analysis of ATF4 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-37, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of ATF4 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1612-37, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: PC-12 cell lysate
Lane 3: HL-60 cell lysate
Lane 4: K562 cell lysate
Lane 5: human lung carcinoma tissue lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant ATF4 Monoclonal Antibody (ET1612-37)

Immunogen

Synthetic peptide within n terminal of human atf4.

Host

Rabbit

Positive Control

Hela cell lysate, PC-12 cell lysate, HL-60 cell lysate, K562 cell lysate, human lung carcinoma tissue lysate, N2A, human liver carcinoma tissue, human prostate carcinoma tissue, human skin tissue, human breast carcinoma tissue, human stomach carcinoma tissue, human small intestine tissue, human colon carcinoma tissue, Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SD20-92

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

55 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

ATF4

SYNONYMS

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

SEQUENCE SIMILARITIES

Belongs to the bZIP family.

POST-TRANSLATIONAL MODIFICATION

Ubiquitinated by SCF(BTRC) in response to mTORC1 signal, followed by proteasomal degradation and leading to down-regulate expression of SIRT4. Interaction with EP300/p300 inhibits ubiquitination by SCF(BTRC).; Phosphorylation at Ser-245 by RPS6KA3/RSK2 in osteoblasts enhances transactivation activity and promotes osteoblast differentiation. 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 (By similarity). Phosphorylation is promoted by mTORC1 (By similarity). Phosphorylation at Ser-215 by CK2 decreases its stability. Phosphorylated by NEK6.; Hydroxylated by PHD3, leading to decreased protein stability.

SUBCELLULAR LOCATION

Cytoplasm, Cell membrane, Nucleus.

FUNCTION

Eukaryotic gene transcription is regulated by sequence-specific transcription factors which bind modular cis-acting promoter and enhancer elements. The cAMP response element (CRE), one of the best studied of such elements, consists of the palindromic octanucleotide TGACGTCA. Several CRE binding proteins have been identified within the ATF/CREB family, the best characterized of which include CREB-1, CREB-2 (also designated ATF-4), ATF-1, ATF-2 and ATF-3. These proteins share highly related COOH terminal leucine zipper dimerization and basic DNA binding domains but are highly divergent in their amino terminal domains. Although each of the ATF/CREB proteins appear capable of binding CRE in its homodimeric form, certain of these also bind as heterodimers, both within the ATF/CREB family and even with members of the AP-1 transcription factor family.