Recombinant Anti-ALKBH1 antibody [EPR19215] (ab195376)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19215] to ALKBH1
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-ALKBH1 antibody [EPR19215]
See all ALKBH1 primary antibodies -
Description
Rabbit monoclonal [EPR19215] to ALKBH1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Sheep, Goat, Horse, Guinea pig, Cow, Cat, Dog, Non human primates -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Jurkat, K562, A549, A431, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; human fetal kidney, colon and fetal liver lysates; mouse brain and spleen lysates; rat spleen lysate. IP: K562 whole cell lysate.
-
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
-
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...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19215 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab195376 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. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
Milk recommended as blocking agent. |
IP |
1/30.
|
Notes |
---|
WB
1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa). Milk recommended as blocking agent. |
IP
1/30. |
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
- Entrez Gene: 211064 Mouse
- Entrez Gene: 362766 Rat
- Omim: 605345 Human
- SwissProt: Q13686 Human
- SwissProt: P0CB42 Mouse
- Unigene: 94542 Human
- Unigene: 26840 Mouse
-
Alternative names
- ABH antibody
- ABH1 antibody
- alkB antibody
see all
Images
-
All lanes : Anti-ALKBH1 antibody [EPR19215] (ab195376) at 1/1000 dilution
Lane 1 : A549 (Human lung carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate
Lane 2 : A549 (Human lung carcinoma epithelial cell) transfected with siRNA specifically targeting ALKBH1 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 44 kDa
Observed band size: 44 kDa
Exposure time: 120 secondsBlocking and diluting buffer: 5% NFDM/TBST
-
All lanes : Anti-ALKBH1 antibody [EPR19215] (ab195376) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : ALKBH1 knockout HAP1 whole cell lysate
Lane 3 : A549 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 44 kDaLanes 1 - 3: Merged signal (red and green). Green - ab195376 observed at 43 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab195376 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. Ab195376 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 [EPR19215] (ab195376) at 1/1000 dilution
Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : A431 (Human epidermoid carcinoma cell line) 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: 44 kDa
Observed band size: 44 kDa
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-ALKBH1 antibody [EPR19215] (ab195376) at 1/1000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human colon lysate
Lane 3 : Human fetal liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 44 kDa
Observed band size: 44 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
-
All lanes : Anti-ALKBH1 antibody [EPR19215] (ab195376) at 1/1000 dilution
Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 5 : Mouse brain lysate
Lane 6 : Mouse spleen lysate
Lane 7 : Rat spleen 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: 44 kDa
Observed band size: 44 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-4: 30 seconds Lane 5-7: 3 minutes.
-
ALKBH1 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate with ab195376 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab195376 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: K562 whole cell lysate 10µg (Input).
Lane 2: ab195376 IP in K562 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab195376 in K562 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (6)
ab195376 has been referenced in 6 publications.
- Cai GP et al. Alkbh1-mediated DNA N6-methyladenine modification regulates bone marrow mesenchymal stem cell fate during skeletal aging. Cell Prolif N/A:e13178 (2022). WB ; Mouse . PubMed: 35018683
- Rashad S et al. Codon Usage and mRNA Stability are Translational Determinants of Cellular Response to Canonical Ferroptosis Inducers. Neuroscience 501:103-130 (2022). WB ; Rat . PubMed: 35987429
- Luo L et al. DNA 6mA Demethylase ALKBH1 Orchestrates Fatty Acid Metabolism and Suppresses Diet-induced Hepatic Steatosis. Cell Mol Gastroenterol Hepatol N/A:N/A (2022). WB, ChIP ; Mouse . PubMed: 36058506
- Rashad S et al. The stress specific impact of ALKBH1 on tRNA cleavage and tiRNA generation. RNA Biol 17:1092-1103 (2020). PubMed: 32521209
- Timms RT et al. A glycine-specific N-degron pathway mediates the quality control of protein N-myristoylation. Science 365:N/A (2019). PubMed: 31273098
- Wu L et al. Association of N6-methyladenine DNA with plaque progression in atherosclerosis via myocardial infarction-associated transcripts. Cell Death Dis 10:909 (2019). WB, IHC-Fr ; Human, Mouse . PubMed: 31797919