Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2846(2)] to RPA32/RPA2 (phospho T21)
- Suitable for: Dot blot, WB, ELISA
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Overview
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Product name
Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)]
See all RPA32/RPA2 primary antibodies -
Description
Rabbit monoclonal [EPR2846(2)] to RPA32/RPA2 (phospho T21) -
Host species
Rabbit -
Specificity
ab109394 only detects RPA32/RPA2 phosphorylated at threonine 21. The antibody has been tested with milk (5%) and BSA (2%) as blocking reagents, but it provided better results with 5% milk.
This antibody cross-reactivates with non-phospho substrate in specific situations.
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Tested Applications & Species
See all applications and species dataApplication Species Dot HumanELISA Recombinant fragmentWB MouseRatHuman -
Immunogen
This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa whole cell lysate treated with Calyculin A or Camptothecin. Dot Blot: RPA32/RPA2 (pT21) phospho peptide.
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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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR2846(2) -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/5000 dilution
Lane 1 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 29 kDa
Observed band size: 32,36 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
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Direct ELISA antigen dose-response curve using purified ab109394.
Primary antibody: ab109394 (0-1000nweg/ml).
Antigens: P1: Human RPA32/RPA2 (phospho T21) at 10ng/ml and 1ng/ml; NP: non-phospho at 10ng/ml and 1 ng/ml.
Secondary antibody: Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at 1/2500 dilution.
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Dot blot analysis of human c-Myb RPA32/RPA2 (pT21) phospho peptide (Lane 1) and RPA32/RPA2 non-phospho peptide (Lane 2) labeling RPA32/RPA2 (phospho T21) with purified ab109394 at a dilution of 1/5000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/2500.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/2000 dilution
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysates
Lane 2 : C6 (Rat glial tumor glial cell) treated with 100nM calyculin A for 60 minutes whole cell lysates
Lane 3 : C6 (Rat glial tumor glial cell) treated with 100nM calyculin A for 60 minutes whole cell lysates.Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 29 kDa
Observed band size: 32,36 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration:5% NFDM/TBST.
Exposure time: 110 seconds.
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All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/2000 dilution
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100ng/ml calyculin A for 30 minutes whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100ng/ml calyculin A for 60 3minutes whole cell lysates.Then the membrane was incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 29 kDa
Observed band size: 32,36 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds.
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All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/50000 dilution
Lane 1 : HeLa whole cell lysate - untreated
Lane 2 : HeLa whole cell lysate - treated with Calyculin A
Lane 3 : HeLa whole cell lysate - treated with Calyculin A and Alkaline Phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 29 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Dot blot analysis of RPA32/RPA2 (pT21) phospho peptide (lane 1) and RPA32/RPA2 non-phospho peptide (lane 2) labelling RPA32/RPA2 (phospho T21) with ab109394 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-RPA32/RPA2 (phospho T21) antibody [EPR2846(2)] (ab109394) at 1/50000 dilution
Lane 1 : HeLa cell lysates, untreated
Lane 2 : HeLa cell lysates, treated with Camptothecin
Lysates/proteins at 10 µg per lane.
Predicted band size: 29 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?
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