Recombinant Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free (ab256316)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3664(2)-25] to HIF Prolyl Hydroxylases - BSA and Azide free
- Suitable for: IHC-P, IP, Flow Cyt (Intra), WB
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-HIF Prolyl Hydroxylases antibody [EPR3664(2)-25] - BSA and Azide free
See all HIF Prolyl Hydroxylases primary antibodies -
Description
Rabbit monoclonal [EPR3664(2)-25] to HIF Prolyl Hydroxylases - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, IP, Flow Cyt (Intra), WBmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- IHC-P: Human cerebrum and lung cancer tissue Flow Cyt (intra): HEK-293 cells IP: HEK-293 whole cell lysate WB: HEK-293 and HeLa whole cell lysates
-
General notes
ab256316 is the carrier-free version of ab255733.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3664(2)-25 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab256316 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 |
---|---|---|
IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
|
IP |
Use at an assay dependent concentration.
|
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
|
WB |
Use at an assay dependent concentration. Detects a band of approximately 47 kDa (predicted molecular weight: 57 kDa).
|
Notes |
---|
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 47 kDa (predicted molecular weight: 57 kDa). |
Target
-
Function
Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF1A at 'Pro-402' and 'Pro-564'. May function as a cellular oxygen sensor and, under normoxic conditions, may target HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. -
Tissue specificity
Widely expressed with highest levels in adult pancreas, heart, skeletal muscle, brain, placenta, kidney and adrenal gland. Expressed at lower levels in epiphyseal cartilage and in fibroblasts. -
Sequence similarities
Contains 2 EF-hand domains.
Contains 1 Fe2OG dioxygenase domain. -
Post-translational
modificationsGlycosylated. -
Cellular localization
Endoplasmic reticulum membrane. - Information by UniProt
-
Database links
- Entrez Gene: 54681 Human
- SwissProt: Q9NXG6 Human
- Unigene: 654944 Human
-
Alternative names
- EGLN4 antibody
- FLJ20262 antibody
- HIF prolyl hydroxylase PH4 antibody
see all
Images
-
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HIF Prolyl Hydroxylases with ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human cerebrum (PMID: 17726031, 12163023). The section was incubated with ab255733 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 Tris-EDTA 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733 ).
-
Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling HIF Prolyl Hydroxylases with ab255733 at 1/250 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human lung cancer (PMID: 24382781). The section was incubated with ab255733 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 Tris-EDTA 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733 ).
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HEK-293 (human embryonic kidney epithelial cell) cells labelling HIF Prolyl Hydroxylases with ab255733 at 1/400 dilution (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733).
-
HIF Prolyl Hydroxylases was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10μg with ab255733 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab255733 1/1000 dilution (0.4 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.
Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10μg
Lane 2: ab255733 IP in HEK-293 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab255733 in HEK-293 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab255733 ).
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
-
Datasheet download
Certificate of Compliance
References (0)
ab256316 has not yet been referenced specifically in any publications.