Recombinant Anti-RBM3 antibody [EPR6061(2)] (ab134946)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6061(2)] to RBM3
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RBM3 antibody [EPR6061(2)]
See all RBM3 primary antibodies -
Description
Rabbit monoclonal [EPR6061(2)] to RBM3 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
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Positive control
- ICC/IF: NIH/3T3 cells; IHC-P: Rat and mouse liver, and human astrocytoma tissues; Flow Cyt (intra): HepG2 cells; WB: Jurkat, HeLa, Neuro-2a, HEK-293T, LnCap, NIH/3T3 and PC-12 whole cell lysates.
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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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 1.50 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6061(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab134946 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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Flow Cyt (Intra) |
1/20.
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WB | (3) |
1/1000 - 1/10000. Predicted molecular weight: 17 kDa.Can be blocked with Human RBM3 peptide (ab188553).
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IHC-P |
1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100.
For unpurified use at 1/250 - 1/500. |
Notes |
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Flow Cyt (Intra)
1/20. |
WB
1/1000 - 1/10000. Predicted molecular weight: 17 kDa.Can be blocked with Human RBM3 peptide (ab188553). |
IHC-P
1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100. For unpurified use at 1/250 - 1/500. |
Target
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Function
Cold-inducible mRNA binding protein that enhances global protein synthesis at both physiological and mild hypothermic temperatures. Reduces the relative abundance of microRNAs, when overexpressed. Enhances phosphoryaltion of translation initiation factors and active polysome formation. -
Sequence similarities
Contains 1 RRM (RNA recognition motif) domain. -
Post-translational
modificationsArg-105 is dimethylated, probably to asymmetric dimethylarginine.
Phosphorylated. -
Cellular localization
Nucleus. Cytoplasm. Cell projection > dendrite. Localizes in mRNA granules in dentrites. - Information by UniProt
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Database links
- Entrez Gene: 5935 Human
- Entrez Gene: 19652 Mouse
- Entrez Gene: 114488 Rat
- Omim: 300027 Human
- SwissProt: P98179 Human
- SwissProt: O89086 Mouse
- SwissProt: Q925G0 Rat
- Unigene: 301404 Human
see all -
Alternative names
- 2600016C11Rik antibody
- IS1 RNPL antibody
- MGC105811 antibody
see all
Images
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All lanes : Anti-RBM3 antibody [EPR6061(2)] (ab134946) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : RBM3 Knockout HEK-293T cell lysate
Lane 3 : Hela cell lysate
Lane 4 : LnCap cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 17 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab134946 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab134946 was shown to react with RBM3 in wild-type HEK-293T cells in Western blot with loss of signal observed in RBM3 knockout cell line ab266663 (RBM3 knockout cell lysate ab258170). Wild-type HEK-293T and RBM3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab134946 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-RBM3 antibody [EPR6061(2)] (ab134946) at 1/1000 dilution (Purified)
Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 17 kDaWe are unsure how to define the extra bands.
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Immunocytochemistry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling RBM3 with Purified ab134946 at 1/100 dilution (1.32 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 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBM3 antibody [EPR6061(2)] (ab134946)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling RBM3 with Purified ab134946 at 1/300 dilution (0.44 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling RBM3 with Purified ab134946 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBM3 antibody [EPR6061(2)] (ab134946)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling RBM3 with Purified ab134946 at 1/300 dilution (0.44 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBM3 antibody [EPR6061(2)] (ab134946)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human astrocytoma tissue sections labeling RBM3 with Purified ab134946 at 1/300 dilution (0.44 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (11)
ab134946 has been referenced in 11 publications.
- Lemieux P et al. Angiostatic freeze or angiogenic move? Acute cold stress prevents angiokine secretion from murine myotubes but primes primary endothelial cells for greater migratory capacity. Front Physiol 13:975652 (2022). PubMed: 36324307
- Sertel SM et al. The mRNA-Binding Protein RBM3 Regulates Activity Patterns and Local Synaptic Translation in Cultured Hippocampal Neurons. J Neurosci 41:1157-1173 (2021). PubMed: 33310754
- Dong W et al. The RNA-binding protein RBM3 promotes cell proliferation in hepatocellular carcinoma by regulating circular RNA SCD-circRNA 2 production. EBioMedicine 45:155-167 (2019). PubMed: 31235426
- Yang HJ et al. Cold-inducible protein RBM3 mediates hypothermic neuroprotection against neurotoxin rotenone via inhibition on MAPK signalling. J Cell Mol Med 23:7010-7020 (2019). PubMed: 31436914
- Lei BB et al. Mild Hypothermia Prevents NO-Induced Cytotoxicity in Human Neuroblastoma Cells Via Induction of COX-2. J Mol Neurosci N/A:N/A (2018). PubMed: 30471009