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Product Info Summary
| SKU: | P00257 |
|---|---|
| Size: | 100ug |
| Reactive Species: | Human, Mouse |
| Host: | Rabbit |
| Application: | ELISA, IHC, WB |
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Product info
Product Name
Anti-Phospho-E2F-1 S364 Antibody
SKU/Catalog Number
P00257
Size
100ug
Form
Liquid (sterile filtered)
Description
Boster Bio Anti-Phospho-E2F-1 S364 Antibody (Catalog # P00257). Tested in ELISA, IHC, WB applications. This antibody reacts with Human, Mouse.
Storage & Handling
Store vial at -20°C prior to opening. Aliquot contents and freeze at -20°C 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 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening. (Ship on dry ice.)
Cite This Product
Anti-Phospho-E2F-1 S364 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # P00257)
Host
Rabbit
Contents
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 0.01% (w/v) Sodium Azide
Clonality
Polyclonal
Isotype
IgG
Immunogen
This affinity purified 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.
Cross-reactivity
No cross reactivity with other proteins.
Reactive Species
P00257 is reactive to E2F1 in Human, Mouse
Observed Molecular Weight
42 kDa
Calculated molecular weight
46.9 kDa
Background of E2F1
E2F-1 (also known as transcription factor E2F-1) is a transcription activator that binds DNA cooperatively with DP proteins through the E2 recognition site, 5'-TTTC[CG]CGC-3'. The E2F family plays a crucial role in the control of cell cycle and action of tumor suppressor proteins and is also a target of the transforming proteins of small DNA tumor viruses. The E2F proteins contain several domains conserved through evolution that are found in most members of the family. These domains include a DNA binding domain, a dimerization domain, a transactivation domain enriched in acidic amino acids, and a tumor suppressor protein association domain which is embedded within the transactivation domain. This protein and two other members, E2F2 and E2F3, have an additional cyclin binding domain. This protein binds preferentially to retinoblastoma protein pRB in a cell-cycle dependent manner. It can mediate both cell proliferation and p53-dependent/independent apoptosis as well as it can block adipocyte differentiation by binding to specific promoters repressing CEBPA binding to its target gene promoters. Anti-E2F-1 pS364 Antibody is useful for researchers involved in transcription factor and DNA binding research.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
P00257 is guaranteed for ELISA, IHC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
ELISA: 1:20,000 - 1:100,000
IHC: 2 mg/ml - 20 µg/ml
WB: 1:250 - 1:2,000
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.
Validation Images & Assay Conditions
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Boster's Affinity Purified anti- E2F-1 pS364 antibody was used at a 10 µg/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. Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.
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Western blot using Boster's Affinity Purified anti-E2F-1 pS364 antibody shows detection of a band at ~47 kDa corresponding to phosphorylated E2F-1 in induced cell lysates. Panel A shows reactivity using a control antibody reactive to all forms of E2F (arrowheads). Panel B shows specific reactivity against phosphorylated E2F-1 (arrowheads) using our anti-E2F-1 pS364 antibody. Lysates are as follows: CRE/E2F-1 are CRE cells derived from mouse NIH3T3 line transfected with human E2F-1, NIH-3T3 used as a negative control, and MDA-MB-231 cells are a human breast cancer line. As indicated each lysate was prepared from untreated cells and cells treated with 2 µM Doxorubicin used as a DNA damaging agent. In addition the MDA-MB-231 cells were also treated with genistein, a mild DNA damaging agent. The figure shows the same membrane first probed with the anti-E2F-1 pS364 antibody used at a 1:250 dilution, then stripped and re-probed with the pan E2F antibody used as a positive control. The positive control antibody clearly shows an E2F-1 band in all human cell lines, but not mouse cells. Treatment with doxorubicin increases the expression of E2F-1 as shown in Panel A. After film development, images were overlapped to confirm that specific anti-E2F-1 pS364 staining shown 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 611-103-122 for 45 min at room temperature for ECL detection. Personal Communication, XiaoHe Yang, Univ. Oklahoma.
Specific Publications For Anti-Phospho-E2F-1 S364 Antibody (P00257)
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5 Customer Q&As for Anti-Phospho-E2F-1 S364 Antibody
Question
Can you help my question with product P00257, anti-Phospho-E2F-1 S364 antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2020-03-26
Answer
It is not recommended storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free P00257 anti-Phospho-E2F-1 S364 antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2020-03-26
Question
Do you have a BSA free version of anti-Phospho-E2F-1 S364 antibody P00257 available?
Verified Customer
Verified customer
Asked: 2020-01-13
Answer
Thank you for your recent telephone inquiry. I can confirm that some lots of this anti-Phospho-E2F-1 S364 antibody P00257 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2020-01-13
Question
I appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for pancreas skin using anti-Phospho-E2F-1 S364 antibody P00257. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-12-03
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-12-03
Question
I see that the anti-Phospho-E2F-1 S364 antibody P00257 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-06-07
Answer
You can find protocols for WB on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to support@bosterbio.com
Boster Scientific Support
Answered: 2019-06-07
Question
See attached the WB image, lot number and protocol we used for pancreas skin using anti-Phospho-E2F-1 S364 antibody P00257. Please let me know if you require anything else.
G. Parker
Verified customer
Asked: 2017-08-16
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2017-08-16
