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ATF4 Recombinant Rabbit Monoclonal Antibody [SD20-92]

Usd: 205

Specification

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Catalog# ET1612-37

ATF4 Recombinant Rabbit Monoclonal Antibody [SD20-92]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

ATF4 Recombinant Rabbit Monoclonal Antibody [SD20-92]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human ATF4 aa 1-220 / 351.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P

Molecular Weight

Predicted band size: 39 kDa

Positive Control

HeLa cell lysate, HL-60 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, Neuro-2a, human colon carcinoma tissue, human liver carcinoma tissue, human thyroid tissue, human prostate carcinoma tissue, human skin tissue, human breast carcinoma tissue, human stomach carcinoma tissue, human small intestine tissue, mouse colon tissue.

Conjugation

unconjugated

Clone Number

SD20-92

RRID

AB_3070110

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:100-1:200

  • IF-Tissue

  • 1:200-1:500

  • IHC-P

  • 1:200-1:1,000

Target

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.

Background References

1. Zhu H et al. Activating transcription factor 4 promotes esophageal squamous cell carcinoma invasion and metastasis in mice and is associated with poor prognosis in human patients. PLoS One 9:e103882 (2014).

2. Lenna S et al. HLA-B35 and dsRNA induce endothelin-1 via activation of ATF4 in human microvascular endothelial cells. PLoS One 8:e56123 (2013).

Sequence Similarity

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

Nucleus, Nucleus speckle, Cell membrane, Cytoplasm, Centrosome.

UNIPROT

SwissProt: P18848 Human

SwissProt: Q06507 Mouse

SwissProt: Q9ES19 Rat

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

Expand

Images

  • Western blot analysis of ATF4 on different lysates with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HL-60 cell lysate (20 µg/Lane)<br />Lane 3: Mouse testis tissue lysate (40 µg/Lane)<br />Lane 4: Rat testis tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 39 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ATF4 on different lysates with Rabbit anti-ATF4 antibody (ET1612-37) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HL-60 cell lysate (20 µg/Lane)
    Lane 3: Mouse testis tissue lysate (40 µg/Lane)
    Lane 4: Rat testis tissue lysate (40 µg/Lane)

    Predicted band size: 39 kDa
    Observed band size: 55 kDa

    Exposure time: 1 minute; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-37) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of Neuro-2a cells labeling ATF4 with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Neuro-2a cells labeling ATF4 with Rabbit anti-ATF4 antibody (ET1612-37) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATF4 antibody (ET1612-37) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/1,000 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (ET1612-37) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/1,000 dilution for 1 hour 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 liver carcinoma tissue with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 liver carcinoma tissue with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 thyroid tissue with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 thyroid tissue with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 mouse colon tissue with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 mouse colon tissue with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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 rat<br />colon tissue with Rabbit anti-ATF4 antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1612-37" style="font-weight: bold;text-decoration: underline;">ET1612-37</a>) at 1/200 dilution for 1 hour 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 rat
    colon tissue with Rabbit anti-ATF4 antibody (ET1612-37) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-37) at 1/200 dilution for 1 hour 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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