Recombinant Anti-ALKBH1 antibody [EPR6176] (ab126596)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6176] to ALKBH1
- Suitable for: WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-ALKBH1 antibody [EPR6176]
See all ALKBH1 primary antibodies -
Description
Rabbit monoclonal [EPR6176] to ALKBH1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-Pmore 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
- WB: Jurkat, K562 and A549 cell lysates. IHC-P: Human kidney and liver tissues. IP: Jurkat and K562 cell lysates.
-
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 2.80 x 10 -11 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...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6176 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab126596 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 | (1) |
1/1000 - 1/10000. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa).
|
IP |
1/10 - 1/100.
|
|
IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
Notes |
---|
WB
1/1000 - 1/10000. Detects a band of approximately 43 kDa (predicted molecular weight: 43 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
-
Function
Dioxygenase that repairs alkylated single-stranded DNA and RNA containing 3-methylcytosine by oxidative demethylation. Requires molecular oxygen, alpha-ketoglutarate and iron. May have a role in placental trophoblast lineage differentiation (By similarity). Has DNA lyase activity and introduces double-stranded breaks at abasic sites. Cleaves both single-stranded DNA and double-stranded DNA at abasic sites, with the greatest activity towards double-stranded DNA with two abasic sites. DNA lyase activity does not require alpha-ketoglutarate and iron. -
Tissue specificity
Ubiquitous. -
Sequence similarities
Belongs to the alkB family.
Contains 1 Fe2OG dioxygenase domain. -
Cellular localization
Mitochondrion. Nucleus. Mainly localizes in euchromatin, largely excluded from heterochromatin and nucleoli. - Information by UniProt
-
Database links
- Entrez Gene: 8846 Human
- Omim: 605345 Human
- SwissProt: Q13686 Human
- Unigene: 94542 Human
-
Alternative names
- ABH antibody
- ABH1 antibody
- alkB antibody
see all
Images
-
All lanes : Anti-ALKBH1 antibody [EPR6176] (ab126596) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : ALKBH1 knockout HAP1 whole cell lysate
Lane 3 : K562 whole cell lysate
Lane 4 : A549 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 43 kDa
Observed band size: 43 kDaLanes 1 - 4: Merged signal (red and green). Green - ab126596 observed at 43 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab126596 was shown to specifically react with in wild-type HAP1 cells as signal was lost in ALKBH1 knockout cells. Wild-type and ALKBH1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab126596 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 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 1/20000 dilution for 1 hour at room temperature before imaging.
-
All lanes : Anti-ALKBH1 antibody [EPR6176] (ab126596) at 1/10000 dilution (purified)
Lane 1 : Jurkat whole cell lysate
Lane 2 : K562 whole cell lysate
Lane 3 : A549 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDa
Blocking and dilution buffer: 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling ALKBH1 with purified ab126596 at a dilution of 1/50. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
-
ab126596 (purified) at a dilution of 1/30 immunoprecipitating ALKBH1 in Jurkat whole cell lysate.
Lane 1 (input): Jurkat whole cell lysate (10µg)
Lane 2 (+): ab126596 + Jurkat whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126596 in Jurkat whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
-
ab126596 (purified) at a dilution of 1/30 immunoprecipitating ALKBH1 in K562 whole cell lysate.
Lane 1 (input): K562 whole cell lysate (10µg)
Lane 2 (+): ab126596 + K562 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126596 in K562 whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
-
All lanes : Anti-ALKBH1 antibody [EPR6176] (ab126596) at 1/1000 dilution (unpurified)
Lane 1 : Jurkat cell lysate
Lane 2 : K562 cell lysate
Lane 3 : A549 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling ALKBH1 with unpurified ab126596 at a dilution of 1/50.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (4)
ab126596 has been referenced in 4 publications.
- Geng Q et al. Comprehensive Analysis of the Prognostic Value and Immune Infiltrates of the Three-m5C Signature in Colon Carcinoma. Cancer Manag Res 13:7989-8002 (2021). PubMed: 34707405
- Guo Y et al. DNA N6-methyladenine modification in hypertension. Aging (Albany NY) 12:6276-6291 (2020). PubMed: 32283543
- Sheng X et al. DNA N6-Methyladenine (6mA) Modification Regulates Drug Resistance in Triple Negative Breast Cancer. Front Oncol 10:616098 (2020). PubMed: 33614498
- Li Y et al. Expression of Demethylase Genes, FTO and ALKBH1, Is Associated with Prognosis of Gastric Cancer. Dig Dis Sci N/A:N/A (2019). PubMed: 30637548