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
Overview
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Product name
Anti-ALKBH1 antibody [EPR19215]
See all ALKBH1 primary antibodies -
Description
Rabbit monoclonal [EPR19215] to ALKBH1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IP HumanWB MouseRatHuman -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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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.
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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
EPR19215 -
Isotype
IgG -
Research areas
Images
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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.
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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.
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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.
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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.
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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.
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