Kerafast抗體Anti-Puromycin [3RH11]

背景介紹：

貨號(hao)：EQ0001，嘌呤霉(mei)素單克隆抗(kang)體(ti)提供了一種(zhong)非放射性方法來測量與嘌呤霉(mei)素孵(fu)育的細(xi)胞(bao)或組織切(qie)片中的整體(ti)蛋白質(zhi)合(he)成（mRNA 翻(fan)譯）速率，或體(ti)內用嘌呤霉(mei)素處理(li)的動(dong)物。

特色：

* 允許直(zhi)接使用標準(zhun)免疫化學方法對翻(fan)譯進行簡單(dan)的評估(gu)和量化

* 傳統脈沖追蹤方法的(de)有利替代方法，后者依賴于放射性氨(an)基酸(suan)標記

* 與蛋白質印跡和(he) ELISA 應(ying)用兼容

* 使(shi)用 Absolute Antibody 的重組平臺制造，具(ju)有(you)來自(zi)雜交瘤 3RH11 的可變區（即特異性）

嘌呤霉素(su)是一種氨基(ji)核苷類(lei)抗生素(su)，來源于白化鏈霉菌(jun)，在核糖體中發生翻譯過程中導致鏈過早終止(zhi)。該分(fen)(fen)(fen)子的(de)(de)(de)一部分(fen)(fen)(fen)類(lei)似于氨酰化 tRNA 的(de)(de)(de) 3' 端，使其可用于蛋(dan)白質(zhi)翻譯分(fen)(fen)(fen)析(xi)。用于監測(ce)(ce)蛋(dan)白質(zhi)合成的(de)(de)(de)經典脈(mo)沖追(zhui)蹤或淹沒劑量(liang)方(fang)法(fa)依賴于放射(she)性蛋(dan)氨酸和（半）氨酸標記的(de)(de)(de)測(ce)(ce)量(liang)。使用嘌呤霉素(su)免(mian)疫檢測(ce)(ce)進行分(fen)(fen)(fen)析(xi)是放射(she)性氨基(ji)酸標記的(de)(de)(de)一種有利替(ti)代方(fang)法(fa)，并且允許使用標準免(mian)疫化學方(fang)法(fa)直接(jie)評估/量(liang)化翻譯

EQ0001，This monoclonal antibody to puromycin provides a non-radioactive method to measure rates of global protein synthesis (mRNA translation) in cells or tissue slices incubated with puromycin, or animals treated with puromycin in vivo.

Highlights:

* Allows for the simple evaluation and quantification of translation directly using standard immunochemical methods

* Advantageous alternative to traditional pulse-chase methods, which rely on radioactive amino acid labeling

* Compatible with Western Blot and ELISA applications

* Manufactured using Absolute Antibody’s Recombinant Platform with variable regions (i.e., specificity) from the hybridoma 3RH11

Puromycin is an aminonucleoside antibiotic, derived from the Streptomyces alboniger bacterium, that causes premature chain termination during translation taking place in the ribosome. Part of the molecule resembles the 3' end of the aminoacylated tRNA, making it useful for protein translation analysis. Classical pulse-chase or flooding dose methods used to monitor protein synthesis rely on the measurement of radioactive methionine and cysteine labels. Analysis using puromycin immunodetection is an advantageous alternative to radioactive amino acid labeling, and allows for the evaluation/quantification of translation directly using standard immunochemical methods

Product Type:	Antibody
Name:	Anti-Puromycin (3RH11)
Host:	Mouse
Isotype:	IgG1 kappa
Clonality:	Monoclonal
Clone Name:	3RH11
Specificity:	This antibody recognizes puromycin.
Immunogen:	puromycin hydrochloride
Format:	Liquid
Purification Method:	Protein G purified
Buffer:	PBS with 0.02% Proclin 300
Tested Applications:	Western blotting (1:1,000), ELISA and Immunofluorescence microscopy.
Storage:	+4C (short-term), -20C (long-term); Avoid repeated freeze/thaw cycles.
Shipped:	Cold packs

數據展示(shi)：

(A) C2C12 myoblasts were starved of serum and leucine for 2 hr and then IGF-1 and leucine were added to the medium of some of the cells for 45 min. Puromycin (1uM) was added to the medium of some of the cells (lanes 3-6) 30 min before harvest. (B) Quantification of western blot analysis from panel A. (C) In the same study, but in a separate set of culture dishes, cells were incubated with [35S]methionine instead of puromycin and incorporation was measured.

A Western blot was run using the same samples where one set was run on the left side of the gel and the other on the right.  The left side was probed with our original monoclonal anti-puromycin antibody and the other side was probed with the recombinant anti-puromycin antibody, both at 1:1,000 dilution. The secondary antibody was used at the same dilution for both sides and they were both exposed for ~40 sec. The first two samples were from skeletal muscle of mice where the first lane is muscle from the control hindlimb and the other is from a hindlimb that had been immobilized with a cast for three days.  The other 3 lanes are from HEK393T cells: the first lane is from cells incubated in complete medium, the middle lane is from cells incubated for 2 hr in medium lacking glucose and serum, and the last lane is cells incubated for 1.5 hr without glucose or serum and then glucose and serum were returned during the last 30 min.  As expected, puromycin incorporation was lower in the immobilized hindlimb compared to the contralateral control hindlimb, and also in cells deprived of glucose and serum compared to cells in complete medium.  Resupplementation partially restored incorporation.

Kerafast抗體Anti-Puromycin [3RH11]已發(fa)表文獻

參考文獻：

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2. Tom Dieck S, Kochen L, Hanus C, Heumüller M, Bartnik I, Nassim-Assir B, Merk K, Mosler T, Garg S, Bunse S, Tirrell DA, Schuman EM. Direct visualization of newly synthesized target proteins in situ. Nat Methods. 2015 May;12(5):411-4.

3. Gantois I, Khoutorsky A, Popic J, Aguilar-Valles A, Freemantle E, Cao R, Sharma V, Pooters T, Nagpal A, Skalecka A, Truong VT, Wiebe S, Groves IA, Jafarnejad SM, Chapat C, McCullagh EA, Gamache K, Nader K, Lacaille JC, Gkogkas CG, Sonenberg N. Metformin ameliorates core deficits in a mouse model of fragile X syndrome. Nat Med. 2017 Jun;23(6):674-677.

4. Rangaraju V, Lauterbach M, Schuman EM. Spatially Stable Mitochondrial Compartments Fuel Local Translation during Plasticity. Cell. 2019 Jan 10;176(1-2):73-84.e15.

5. Hafner AS, Donlin-Asp PG, Leitch B, Herzog E, Schuman EM. Local protein synthesis is a ubiquitous feature of neuronal pre- and postsynaptic compartments. Science. 2019 May 17;364(6441).

6. H?rnberg H, Pérez-Garci E, Schreiner D, et al. Rescue of oxytocin response and social behaviour in a mouse model of autism. Nature. 2020;584(7820):252-256.

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