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Anti-Puromycin antibody [EPR27218-173] (ab315887) is a rabbit monoclonal antibody that is used to detect Puromycin in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

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Images

Immunoprecipitation - Anti-Puromycin antibody [EPR27218-173] (AB315887), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (AB315887), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (AB315887), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (AB315887), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (AB315887), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PICC/IFWBFlow Cyt (Intra)IP
Chemical
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species
Chemical
Dilution info
1/5000
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species
Chemical
Dilution info
1/500
Notes

-

Tested
Tested

Species
Chemical
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Chemical
Dilution info
1/500
Notes

-

Tested
Tested

Species
Chemical
Dilution info
1/30
Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Recommended products

Anti-Puromycin antibody [EPR27218-173] (ab315887) is a rabbit monoclonal antibody that is used to detect Puromycin in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF.



- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR27218-173
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

What is this antibody validated in?


Anti-Puromycin antibody [EPR27218-173] (ab315887) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), .

Recommended positive controls


WB: HeLa treated with 1uM Puromycin for 30min whole, NIH/3T3 treated with1uM Puromycin for 30min whole and C6 treated with 1uM Puromycin for 30min whole lysates.IHC-P: HeLa cell pellet treated with Puromycin (1 uM) for 30min.ICC: HeLa cells after treatment with 1 uM Puromycin for 30 mins.Flow Cyt: HeLa (human cervical adenocarcinoma epithelial cell) treated with 1uM Puromycin for 30min.IP: HeLa treated with 1uM Puromycin for 30min.

Trial sizes available!


Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.Other related products


We have a range of other formats of antibody clone [EPR27218-173] also available for your convenience:
ab315887, Carrier free - Anti-Puromycin antibody [EPR27218-173] - BSA and Azide free ab315888, Alexa Fluor® 647 - Alexa Fluor® 647 Anti-Puromycin antibody [EPR27218-173] ab322729, Alexa Fluor® 488 - Alexa Fluor® 488 Anti-Puromycin antibody [EPR27218-173] ab323262



Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Puromycin is a well-known antibiotic that acts by disrupting protein synthesis. It mimics the structure of an aminoacyl-tRNA leading to premature termination of the growing polypeptide chain during translation. Puromycin is small with a molecular mass of approximately 471 daltons. It interacts mainly within the cytoplasm where it can incorporate into the peptide chain due to its similarity to the 3' end of the aminoacyl-tRNA. Puromycin does not have a specific expression site like endogenous proteins because it is not naturally occurring in animals; rather it is used experimentally.

Biological function summary

The function of puromycin is to interfere with protein synthesis by targeting ribosomes. It binds to the ribosomal A site leading to the release of incomplete polypeptides. Puromycin is not part of a complex but establishes a strong interaction with ribosomes found in prokaryotic and eukaryotic cells. This interaction gives it unique utility in experiments that require inhibition of translation such as studies involving protein turnover and synthesis rates.

Pathways

The action of puromycin strongly impacts protein synthesis pathways. It inhibits the elongation stage of the translation pathway by mimicking an aminoacyl-tRNA. Puromycin does not involve complex pathways itself but its effects can influence other proteins involved in translation such as elongation factors and the ribosomal subunits. This makes puromycin useful in basic scientific research to study how cells control protein synthesis.

Associated diseases and disorders

Puromycin's ability to inhibit translation links it to studies on cancer and neurodegenerative diseases. Researchers use puromycin to explore how disrupting protein synthesis can affect cancer cell growth. It serves as a model for understanding toxins that affect similar pathways in neurodegenerative disorders. Through these studies connections have been made between puromycin's action and proteins involved in these diseases such as oncogenes and neuromodulatory proteins which are key to understanding disease progression and developing potential treatments.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunoprecipitation - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Immunoprecipitation - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Puromycin was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) treated with 1uM Puromycin for 30min whole cell lysate with ab315887 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315887 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) treated with 1uM Puromycin for 30min whole cell lysate

    Lane 2: ab315887 IP in HeLa (human cervical adenocarcinoma epithelial cell) treated with 1uM Puromycin for 30min whole cell lysate

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab315887 in HeLa treated with 1uM Puromycin for 30min whole cell lysate

    All lanes: Immunoprecipitation - Anti-Puromycin antibody [EPR27218-173] (ab315887) at 1/30 dilution

    All lanes: HeLa (human cervical adenocarcinoma epithelial cell) treated with 1uM Puromycin for 30min whole cell lysate with 5% NFDM/TBST

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

    Exposure time: 8s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Puromycin with ab315887 at 1/100 (5.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Negative control: no staining on mouse kidney. The section was incubated with ab315887 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Immunohistochemical analysis of paraffin-embedded Panel A HeLa (human cervical adenocarcinoma epithelial cell) cell pellet treated with Puromycin (1 uM) for 30min; Panel B untreated HeLa cell pellet tissue labeling Puromycin with ab315887 at 1/5000 (0.105 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining on (A) HeLa cell pellet treated with Puromycin (1 uM) for 30min; no staining on (B) untreated HeLa cell pellet. The section was incubated with ab315887 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Puromycin with ab315887 at 1/100 (5.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Negative control: no staining on rat kidney. The section was incubated with ab315887 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Puromycin with ab315887 at 1/100 (5.24 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Negative control: no staining on human kidney. The section was incubated with ab315887 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunocytochemistry/ Immunofluorescence - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Puromycin with ab315887 at 1/500 (1.048 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

    Confocal image showing cytoplasmic staining in HeLa cells after treatment with 1 uM Puromycin for 30 mins, while showing no staining in untreated HeLa cells.

    Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

    Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

  • Western blot - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Western blot - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Puromycin is a naturally occurring aminonucleoside antibiotic that inhibits protein synthesis by ribosome-catalyzed incorporation into the C-terminus of elongating nascent chains, blocking further extension and resulting in premature termination of translation.

    In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) staining at 1/100000 dilution.

    Exposure time: Lane 1-2: 1 second, Lane 3-4: 6 seconds, Lane 5-6: 3 seconds.

    All lanes: Western blot - Anti-Puromycin antibody [EPR27218-173] (ab315887) at 1/1000 dilution

    Lane 1: Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: HeLa treated with 1uM Puromycin for 30min whole cell lysate at 20 µg

    Lane 3: Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lane 4: NIH/3T3 treated with 1uM Puromycin for 30min whole cell lysate at 20 µg

    Lane 5: Untreated C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

    Lane 6: C6 treated with 1uM Puromycin for 30min whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 10-250 kDa, 15 kDa

  • Flow Cytometry (Intracellular) - Anti-Puromycin antibody [EPR27218-173] (ab315887), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Puromycin antibody [EPR27218-173] (ab315887)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) treated with 1uM Puromycin for 30min (Red) / Untreated HeLa (Green) cells labelling Puromycin with ab315887 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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