Recombinant Anti-PDCD4 antibody [EPR3432] (ab79405)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3432] to PDCD4
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-PDCD4 antibody [EPR3432]
See all PDCD4 primary antibodies -
Description
Rabbit monoclonal [EPR3432] to PDCD4 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HAP1, and Jurkat whole cell lysate. IHC-P: Human breast carcinoma tissue. ICC/IF: HeLa cells
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3432 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab79405 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/5000 - 1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
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IHC-P |
1/1000 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
1/500.
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Notes |
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WB
1/5000 - 1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa). |
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/500. |
Target
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Function
Inhibits translation initiation and cap-dependent translation. May excert its function by hindering the interaction between EIF4A1 and EIF4G. Inhibits the helicase activity of EIF4A. Modulates the activation of JUN kinase. Down-regulates the expression of MAP4K1, thus inhibiting events important in driving invasion, namely, MAPK85 activation and consequent JUN-dependent transcription. May play a role in apoptosis. Tumor suppressor. Inhibits tumor promoter-induced neoplastic transformation. Binds RNA. -
Tissue specificity
Up-regulated in proliferative cells. Highly expressed in epithelial cells of the mammary gland. Reduced expression in lung cancer and colon carcinoma. -
Sequence similarities
Belongs to the PDCD4 family.
Contains 2 MI domains. -
Domain
Binds EIF4A1 via both MI domains. -
Post-translational
modificationsPolyubiquitinated, leading to its proteasomal degradation. Rapidly degraded in response to mitogens. Phosphorylation of the phosphodegron promotes interaction with BTRC and proteasomal degradation. -
Cellular localization
Nucleus. Cytoplasm. Shuttles between the nucleus and cytoplasm. Predominantly nuclear under normal growth conditions, and when phosphorylated at Ser-457. Exported from the nucleus in the absence of serum. - Information by UniProt
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Database links
- Entrez Gene: 27250 Human
- Entrez Gene: 18569 Mouse
- Omim: 608610 Human
- SwissProt: Q53EL6 Human
- SwissProt: Q61823 Mouse
- Unigene: 711490 Human
- Unigene: 1605 Mouse
- Unigene: 375091 Mouse
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Alternative names
- Death up-regulated gene protein antibody
- Dug antibody
- H731 antibody
see all
Images
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All lanes : Anti-PDCD4 antibody [EPR3432] (ab79405) at 1/5000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PDCD4 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 51 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab79405 observed at 51 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab79405 was shown to react with PDCD4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261833 (knockout cell lysate ab257278) was used. Wild-type HeLa and PDCD4 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab79405 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling PDCD4 with purified ab79405 at 1/500 dilution (4 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.
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All lanes : Anti-PDCD4 antibody [EPR3432] (ab79405) at 1/5000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : PDCD4 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HEK293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 52 kDaLanes 1 - 4: Merged signal (red and green). Green - ab79405 observed at 52 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab79405 was shown to recognize PDCD4 in wild-type HAP1 cells as signal was lost at the expected MW in PDCD4 knockout HAP1 cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PDCD4 knockout samples were subjected to SDS-PAGE. ab79405 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-PDCD4 antibody [EPR3432] (ab79405) at 1/10000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa
Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands. -
ab79405 at 1/1000 dilution staining PDCD4 in human breast carcinoma by Immunohistochemistry using paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab79405 has been referenced in 10 publications.
- Huang WC et al. The MEK/ERK/miR-21 Signaling Is Critical in Osimertinib Resistance in EGFR-Mutant Non-Small Cell Lung Cancer Cells. Cancers (Basel) 13:N/A (2021). PubMed: 34885115
- Hale BJ et al. Inhibition of germinal vesicle breakdown using IBMX increases microRNA-21 in the porcine oocyte. Reprod Biol Endocrinol 18:39 (2020). PubMed: 32393269
- Patutina OA et al. Mesyl phosphoramidate backbone modified antisense oligonucleotides targeting miR-21 with enhanced in vivo therapeutic potency. Proc Natl Acad Sci U S A 117:32370-32379 (2020). PubMed: 33288723
- Hua R et al. Ssc-miR-21-5p regulates endometrial epithelial cell proliferation, apoptosis and migration via the PDCD4/AKT pathway. J Cell Sci 133:N/A (2020). PubMed: 33097608
- Wäster P et al. Extracellular vesicles released by melanocytes after UVA irradiation promote intercellular signaling via miR21. Pigment Cell Melanoma Res 33:542-555 (2020). PubMed: 31909885
- Chen T et al. Involvement of androgen receptor (AR)/microRNA-21 axis in hypoxia/reoxygenation-induced apoptosis of mouse renal tubular epithelial cells. Am J Transl Res 11:5611-5622 (2019). PubMed: 31632533
- Zhang Y et al. Expression and function of Pdcd4 in mouse endometrium during early pregnancy. Reproduction 155:393-402 (2018). PubMed: 29459402
- Patutina OA et al. miRNases: Novel peptide-oligonucleotide bioconjugates that silence miR-21 in lymphosarcoma cells. Biomaterials 122:163-178 (2017). PubMed: 28126663
- Liang X et al. MicroRNA-16 suppresses the activation of inflammatory macrophages in atherosclerosis by targeting PDCD4. Int J Mol Med 37:967-75 (2016). PubMed: 26936421
- Wright EC et al. MicroRNA-21 and PDCD4 expression during in vitro oocyte maturation in pigs. Reprod Biol Endocrinol 14:21 (2016). PubMed: 27084064