Recombinant Anti-HIF1 beta antibody [EPR23106-151] (ab270520)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23106-151] to HIF1 beta
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-HIF1 beta antibody [EPR23106-151]
See all HIF1 beta primary antibodies -
Description
Rabbit monoclonal [EPR23106-151] to HIF1 beta -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: ChIP,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MDA-MB-231, RAW264.7, 293T, C6, HAP1 and HeLa cell lysates. IHC-P: Human colon tissue: Mouse testis tissue; Rat testis tissue. Flow Cyt (intra): HeLa and NIH/3T3 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.
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. -
Storage buffer
pH: 7.2
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
EPR23106-151 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab270520 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) |
1/600.
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WB |
1/1000. Detects a band of approximately 87 kDa (predicted molecular weight: 87 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/600. |
WB
1/1000. Detects a band of approximately 87 kDa (predicted molecular weight: 87 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Required for activity of the Ah (dioxin) receptor. This protein is required for the ligand-binding subunit to translocate from the cytosol to the nucleus after ligand binding. The complex then initiates transcription of genes involved in the activation of PAH procarcinogens. The heterodimer with HIF1A or EPAS1/HIF2A functions as a transcriptional regulator of the adaptive response to hypoxia. -
Sequence similarities
Contains 1 basic helix-loop-helix (bHLH) domain.
Contains 1 PAC (PAS-associated C-terminal) domain.
Contains 2 PAS (PER-ARNT-SIM) domains. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 405 Human
- Entrez Gene: 11863 Mouse
- Entrez Gene: 25242 Rat
- Omim: 126110 Human
- SwissProt: P27540 Human
- SwissProt: P53762 Mouse
- SwissProt: P41739 Rat
- Unigene: 632446 Human
see all -
Alternative names
- arnT antibody
- ARNT protein antibody
- ARNT_HUMAN antibody
see all
Images
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All lanes : Anti-HIF1 beta antibody [EPR23106-151] (ab270520) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : ARNT knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 87 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab270520 observed at 90 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab270520 was shown to react with HIF1 beta in wild-type HAP1 cells in Western blot with loss of signal observed in ARNT knockout sample.Wild-type HAP1 and ARNT knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5 % milk in TBS-T (0.1 % Tween®) before incubation with ab270520 and ab8245 (Mouse anti-GAPDH antibody [6C5]) 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 beta antibody [EPR23106-151] (ab270520)
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling HIF1 beta with ab270520 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat testis is observed. The section was incubated with ab270520 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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All lanes : Anti-HIF1 beta antibody [EPR23106-151] (ab270520) at 1/1000 dilution
Lane 1 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 3 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 5 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 87 kDa
Observed band size: 87 kDa
Exposure time: 26 secondsBlocking and dilution buffer: 5% NFDM/TBST.
Lysates should be made fresh and used in WB immediately to minimize protein degradation.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 beta antibody [EPR23106-151] (ab270520)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling HIF1 beta with ab270520 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse testis is observed. The section was incubated with ab270520 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling HIF1 beta with ab270520 at 1/600 dilution (0.1 µg) (Red) compared with a Rabbit monoclonal IgG (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, ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 beta antibody [EPR23106-151] (ab270520)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HIF1 beta with ab270520 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human colon is observed. The section was incubated with ab270520 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling HIF1 beta with ab270520 at 1/600 dilution (0.1 µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab270520 has not yet been referenced specifically in any publications.