Anti-Rex1 antibody (ab28141)
Key features and details
- Rabbit polyclonal to Rex1
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Rex1 antibody
See all Rex1 primary antibodies -
Description
Rabbit polyclonal to Rex1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
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Positive control
- Lysate of 293T cells overexpressing Rex1, Mouse Embryonic Stem Cells expressing biotinylated Rex1.
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab28141 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 WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 38 kDa (predicted molecular weight: 32 kDa). ICC/IF Use a concentration of 1 µg/ml. Target
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Function
Involved in self-renewal property of ES cells (By similarity). May be involved in transcriptional regulation. -
Tissue specificity
Expressed in kidney, epidermal keratinocytes, prostate epithelial cells, bronchial and small airway lung epithelial cells (at protein level). Expressed in malignant kidney and several carcinoma cell lines (at protein level). Expressed in embryonic stem cells, kidney, epidermal keratinocytes, prostate epithelial cells, bronchial and small airway lung epithelial cells. Expressed in embryonal carcinomas, seminomas, malignant kidney and several carcinoma cell lines. -
Sequence similarities
Belongs to the krueppel C2H2-type zinc-finger protein family.
Contains 4 C2H2-type zinc fingers. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 132625 Human
- Entrez Gene: 22702 Mouse
- Omim: 614572 Human
- SwissProt: Q96MM3 Human
- SwissProt: P22227 Mouse
- Unigene: 335787 Human
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Alternative names
- hREX-1 antibody
- Reduced Expression 1 Protein antibody
- Reduced expression protein 1 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Rex1 antibody (ab28141)ICC/IF image of ab28141 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28141, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
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Western blot - Anti-Rex1 antibody (ab28141)All lanes : Anti-Rex1 antibody (ab28141) at 0.5 µg/ml
Lane 1 : Lysate of 293T cells overexpressing Rex1
Lane 2 : Lysate of Mouse Embryonic Stem Cells expressing biotinylated Rex1
Lane 3 : Negative control for tagged protein - Lysate of Mouse Embryonic Stem Cells (endogenous protein present)
Lanes 4-7 : Lysate of various passages of Mouse Embryonic Stem Cells expressing biotinylated Rex1
Predicted band size: 32 kDa
Observed band size: 32 kDa
Additional bands at: 34 kDa (possible non-specific binding), 36 kDa (possible non-specific binding), 38 kDa (possible tagged protein)
In this western blot Anti-Rex1 antibody ab28141 recognizes a band at 32kDa corresponding to the endogenous Rex1 protein and a band at 38 kDa corresponding to biotinylated Rex1. The identities of the bands at 34 and 36 kDa are unknown. It is possible that they represent non-specific binding, but it may be that they represent modified forms of the non-biotinylated endogenous Rex1. The biotinylated Rex1 protein contains an N-terminal FLAG and a 25 AA peptide substrate for biotinylation. Therefore, the biotinylated protein is expected to migrate at a slightly higher molecular weight than endogenous Rex1. The large bands at the top of the image are likely to represent non-specific binding. -
Western blot - Anti-Rex1 antibody (ab28141)Anti-Rex1 antibody (ab28141) at 1 µg/ml + 293T Cells Overexpressing Mouse Rex1 at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa, 75 kDa. We are unsure as to the identity of these extra bands.
The REX1 protein in these 293T cells contains an N-terminal FLAG tag. We expect the tagged protein to migrate at a higher molecular weight than endogenous REX1.
Protocols
Datasheets and documents
References (13)
ab28141 has been referenced in 13 publications.
- Bustos F et al. Functional Diversification of SRSF Protein Kinase to Control Ubiquitin-Dependent Neurodevelopmental Signaling. Dev Cell 55:629-647.e7 (2020). PubMed: 33080171
- Liu J et al. Deriving rabbit embryonic stem cells by small molecule inhibitors. Am J Transl Res 11:5122-5133 (2019). PubMed: 31497228
- Bustos F et al. RNF12 X-Linked Intellectual Disability Mutations Disrupt E3 Ligase Activity and Neural Differentiation. Cell Rep 23:1599-1611 (2018). PubMed: 29742418
- Wang F et al. Rlim-Dependent and -Independent Pathways for X Chromosome Inactivation in Female ESCs. Cell Rep 21:3691-3699 (2017). WB, ICC/IF ; Mouse . PubMed: 29281819
- Loos F et al. Xist and Tsix Transcription Dynamics Is Regulated by the X-to-Autosome Ratio and Semistable Transcriptional States. Mol Cell Biol 36:2656-2667 (2016). PubMed: 27528619
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Rex1 antibody (ab28141)ICC/IF image of ab28141 stained mouse embryonic stem cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28141, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
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Western blot - Anti-Rex1 antibody (ab28141)All lanes : Anti-Rex1 antibody (ab28141) at 0.5 µg/ml
Lane 1 : Lysate of 293T cells overexpressing Rex1
Lane 2 : Lysate of Mouse Embryonic Stem Cells expressing biotinylated Rex1
Lane 3 : Negative control for tagged protein - Lysate of Mouse Embryonic Stem Cells (endogenous protein present)
Lanes 4-7 : Lysate of various passages of Mouse Embryonic Stem Cells expressing biotinylated Rex1
Predicted band size: 32 kDa
Observed band size: 32 kDa
Additional bands at: 34 kDa (possible non-specific binding), 36 kDa (possible non-specific binding), 38 kDa (possible tagged protein)
In this western blot Anti-Rex1 antibody ab28141 recognizes a band at 32kDa corresponding to the endogenous Rex1 protein and a band at 38 kDa corresponding to biotinylated Rex1. The identities of the bands at 34 and 36 kDa are unknown. It is possible that they represent non-specific binding, but it may be that they represent modified forms of the non-biotinylated endogenous Rex1. The biotinylated Rex1 protein contains an N-terminal FLAG and a 25 AA peptide substrate for biotinylation. Therefore, the biotinylated protein is expected to migrate at a slightly higher molecular weight than endogenous Rex1. The large bands at the top of the image are likely to represent non-specific binding. -
Western blot - Anti-Rex1 antibody (ab28141)Anti-Rex1 antibody (ab28141) at 1 µg/ml + 293T Cells Overexpressing Mouse Rex1 at 10 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?
Additional bands at: 100 kDa, 75 kDa. We are unsure as to the identity of these extra bands.
The REX1 protein in these 293T cells contains an N-terminal FLAG tag. We expect the tagged protein to migrate at a higher molecular weight than endogenous REX1.
