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E2F-1 [p Ser364] Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NB600-900

Novus Biologicals, part of Bio-Techne
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NB600-900

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

ELISA, Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Specifications

Immunogen

This affinity purified E2F-1 [p Ser364] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near amino acids 350-375 of Human E2F-1. (Uniprot: Q01094)

Reactivity Notes

A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence. Cross reactivity with E2F-1 homologues from other sources has not been determined.

Modification

p Ser364

Specificity

This affinity purified antibody is directed against the phosphorylated form of human E2F-1 at the pS364 residue. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human E2F-1 pS364 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human E2F-1 is minimal by ELISA. The antibody does not cross-react with E2F-1 phosphorylated at other sites. A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence. Cross reactivity with E2F-1 homologues from other sources has not been determined.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This affinity purified antibody is directed against the phosphorylated form of human E2F-1 at the [p Ser364] residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for E2F-1 [p Ser364] Antibody

Western Blot: E2F-1 [p Ser364] Antibody [NB600-900] - Panel A: Control antibody reactive to all forms of E2F (arrowheads). Panel B: Reactivity with phosphorylated E2F-1 (arrows) using E2F-1 [p Ser364] antibody. Lysate cells: CRE/E2F-1 CRE (human E2F-1 transfected mouse NIH3T3); NIH-3T3 (-ve control), & MDA-MB-231. Each lysate prepared from untreated cells & cells with 2 uM Doxorubicin. MDA-MB-231 also treated with genistein. Same membrane probed with E2F-1 [p Ser364] antibody @ 1:250 dilution, then stripped & re-probed with pan E2F antibody (+ve control). +ve control antibody shows an E2F-1 band in all human cell lines, but not mouse cells. Doxorubicin increases expression of E2F-1 (Panel A). Images overlapped to confirm that specific anti-E2F-1 pS364 staining shown in treated human cells in Panel B specifically aligns with E2F-1 staining shown in Panel A. Blots can be processed with HRP conjugated Gt-a-Rabbit IgG MX10 for 45 min at RT for ECL detection.
Immunohistochemistry: E2F-1 [p Ser364] Antibody [NB600-900] - Affinity Purified E2F-1 [p Ser364] antibody was used at a 10 ug/ml to detect nuclear and occasionally cytoplasmic signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. Within the multi-tumor block, the antibody showed variable levels of nuclear staining between individual tumors, with some tumors showing strong staining. This image shows E2F-1 pS364 staining of human breast carcinoma. Tissue was formalin-fixed and paraffin embedded.
Western Blot: E2F-1 [p Ser364] Antibody [NB600-900]

Western Blot: E2F-1 [p Ser364] Antibody [NB600-900]

Western Blot: E2F-1 [p Ser364] Antibody [NB600-900] - NB600-900 Western blot - E2F1 (phospho S364) antibody

Applications for E2F-1 [p Ser364] Antibody

Application
Recommended Usage

ELISA

1:20000-1:100000

Immunohistochemistry

2 mg/ml-20 ug/ml

Western Blot

1:250-1:2000
Application Notes
This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 47 kDa in size corresponding to phosphorylated E2F-1 by western blotting in the appropriate cell lysate or extract. Less than 0.5% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for pS364 of E2F-1.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: E2F-1

E2F's are DNA binding proteins, which associate with negative regulators, such as the retinoblastoma p107 protein, resulting in an altered rate of gene transcription. E2F1 is proposed to be involved in several cellular processes that range from tumor suppressor, cell progression and oncogenesis. E2F1 overexpression can also drive cells into apoptosis.

Long Name

E2F Transcription Factor 1

Alternate Names

E2F1, PBR3, RBAP-1, RBBP-3, RBP3

Entrez Gene IDs

1869 (Human); 13555 (Mouse)

Gene Symbol

E2F1

UniProt

Additional E2F-1 Products

Product Documents for E2F-1 [p Ser364] Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for E2F-1 [p Ser364] Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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