Anti-DNA Polymerase beta antibody (ab26343)
Key features and details
- Rabbit polyclonal to DNA Polymerase beta
- Suitable for: WB, ICC/IF, IHC-P
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-DNA Polymerase beta antibody
See all DNA Polymerase beta primary antibodies -
Description
Rabbit polyclonal to DNA Polymerase beta -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB MouseHuman
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Immunogen
Synthetic peptide corresponding to Human DNA Polymerase beta aa 300 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Images
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Western blot - Anti-DNA Polymerase beta antibody (ab26343)All lanes : Anti-DNA Polymerase beta antibody (ab26343) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : POLB knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 38 kDaLanes 1-3: Merged signal (red and green). Green - ab26343 observed at 38 kDa. Red - loading control, ab7291 observed at 50 kDa.
ab26343 Anti-DNA Polymerase beta antibody was shown to specifically react with DNA Polymerase beta in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261748 (knockout cell lysate ab257096) was used. Wild-type and DNA Polymerase beta knockout samples were subjected to SDS-PAGE. ab26343 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti- Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti- Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-DNA Polymerase beta antibody (ab26343)All lanes : Anti-DNA Polymerase beta antibody (ab26343) at 0.5 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899)
Lane 3 :A-431 whole cell lysate (ab7909)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 38 kDa
Additional bands at: 150 kDa (possible cross reactivity), 48 kDa (possible cross reactivity), 90 kDa (possible cross reactivity) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA Polymerase beta antibody (ab26343)Formaldehyde-fixed human small cell lung cancer tissue stained for DNA Polymerase beta using ab26343 at 1/200 dilution in immunohistochemical analysis.
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Western blot - Anti-DNA Polymerase beta antibody (ab26343)All lanes : Anti-DNA Polymerase beta antibody (ab26343) at 1 µg/ml
Lane 1 :NIH/3T3 whole cell lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 38 kDa
Additional bands at: 120 kDa (possible cross reactivity), 30 kDa (possible cross reactivity) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA Polymerase beta antibody (ab26343) Image courtesy of Human Protein AtlasImage courtesy of Human Protein Atlas ab26343 staining DNA polymerase beta in Human gall bladder, showing a strong nuclear staining pattern in the columnar epithelial cells in contrast with the cells in the lamina propria. Paraffin embedded human gall bladder tissue was incubated with ab26343 (1/150 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab26343 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
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Immunocytochemistry/ Immunofluorescence - Anti-DNA Polymerase beta antibody (ab26343)ICC/IF image of ab26343 stained T47D cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab26343 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
