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phospho-ATF2 (Thr69/71) Polyclonal Antibody

Paraformaldehyde-fixed, paraffin embedded Human Colon carcinoma; 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, followed by secondary and DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon carcinoma; 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, followed by secondary and DAB staining.
Paraformaldehyde-fixed, paraffin-embedded Rat 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, followed by secondary and DAB staining.
Paraformaldehyde-fixed, paraffin-embedded Rat 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, followed by secondary and DAB staining.
Mouse spleen cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ATF2 (Thr69\/71) Polyclonal Antibody(bs-3033R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Mouse spleen cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ATF2 (Thr69\/71) Polyclonal Antibody(bs-3033R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Paraformaldehyde-fixed, paraffin embedded Rat 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human colon cancer; 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human colon cancer; 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\u00b0C for 30min; Antibody incubation with ATF2 (Thr69\/71) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human pancreas); 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\u00b0C for 30min; Incubation with (phospho-ATF2 (Thr69\/71)) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human pancreas); 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\u00b0C for 30min; Incubation with (phospho-ATF2 (Thr69\/71)) Polyclonal Antibody, Unconjugated (bs-3033R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Bioss unconjugated antibody vial

Applications

  • ELISA
  • FCM
  • IHC-P
  • IHC-F
  • IF(IHC-P)
  • IF(IHC-F)
  • IF(ICC)

Reactivity

  • Mouse
  • Rat

Predicted Reactivity

  • Human
  • Cow
  • Horse
  • Rabbit
Overview
Catalog # bs-3033R-100uL
Product Name phospho-ATF2 (Thr69/71) Polyclonal Antibody
Applications
ELISA FCM IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Specificity This antibody is specific for ATF2 in when phosphorylated at both the Thr69 and Thr71 residues
Reactivity Mouse, Rat
Predicted Reactivity Human, Cow, Horse, Rabbit
Specifications
Conjugation Unconjugated
Host Rabbit
Source KLH conjugated synthetic phosphopeptide derived from human ATF2 around the phosphorylation site of Thr69/71
Modification Site Thr69/71
Clonality Polyclonal
Isotype IgG
Concentration 1ug/ul
Purification Purified by Protein A.
Storage Buffer 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 1386
Swiss Prot P15336
Subcellular location Nucleus
Synonyms ATF2 phospho T69 + T71; ATF2 phospho Thr69 + Thr71; p-ATF2 T69 + T71; p-ATF2 Thr69 + Thr71; CREB 2; HB 16;Activating Transcription Factor 2; ATF 2; Atf-2; ATF2 protein; cAMP Response Element Binding Protein 2; cAMP response element binding protein CRE BP1; cAMP-dependent transcription factor ATF-2; cAMP-responsive element-binding protein 2; CRE BP1; CRE-BP; CREB 2; CREB2; CREBP1; Cyclic AMP dependent transcription factor ATF 2; Cyclic AMP-responsive; ATF2_HUMAN.
Background ATF2 is a member of the ATF/CREB family of basic region leucine zipper DNA binding proteins that regulates transcription by binding to a consensus cAMP response element (CRE) in the promoter of various viral and cellular genes. Many of these genes are important in cell growth and differentiation, and in stress and immune responses. ATF2 is a nuclear protein that binds DNA as a dimer and can form dimers with members of the ATF/CREB and Jun/Fos families. It is a stronger activator as a heterodimer with cJun than as a homodimer. Several isoforms of ATF2 arise by differential splicing. The stable native full length ATF2 is transcriptionally inactive as a result of an inhibitory direct intramolecular interaction of its carboxy terminal DNA binding domain with the amino terminal transactivation domain. Following dimerization ATF2 becomes a short lived protein that undergoes ubiquitination and proteolysis, seemingly in a protein phosphatase-dependent mechanism. Stimulation of the transcriptional activity of ATF2 occurs following cellular stress induced by several genotoxic agents, inflammatory cytokines, and UV irradiation. This activation requires phosphorylation of two threonine residues in ATF2 by both JNK/SAP kinase and p38 MAP kinase. ATF2 is abundantly expressed in brain.
Application Dilution
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Frequently asked questions

Which applications is phospho-ATF2 (Thr69/71) Polyclonal Antibody (bs-3033R) validated for, and at what dilution?

bs-3033R is validated for ELISA (1:500-1000); FCM (1:20-100); IHC-P (1:200-400); IHC-F (1:100-500); IF(IHC-P) (1:50-200); IF(IHC-F) (1:50-200); IF(ICC) (1:50-200). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does phospho-ATF2 (Thr69/71) Polyclonal Antibody (bs-3033R) react with?

Tested reactivity: Mouse, Rat. Predicted reactivity (expected from sequence homology but not yet tested): Human, Cow, Horse, Rabbit.

How should bs-3033R be stored?

Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

What immunogen was used to raise phospho-ATF2 (Thr69/71) Polyclonal Antibody (bs-3033R)?

KLH conjugated synthetic phosphopeptide derived from human ATF2 around the phosphorylation site of Thr69/71

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