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AB326948

Anti-DNA Polymerase theta antibody [EPR28988-1] - BSA and Azide free

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Rabbit Recombinant Monoclonal DNA Polymerase theta antibody. Carrier free. Suitable for WB and reacts with Human samples.

View Alternative Names

POLH, POLQ, DNA polymerase theta, DNA polymerase eta

2 Images
Western blot - Anti-DNA Polymerase theta antibody [EPR28988-1] - BSA and Azide free (AB326948)
  • WB

Lab

Western blot - Anti-DNA Polymerase theta antibody [EPR28988-1] - BSA and Azide free (AB326948)

This data was developed using ab326932, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot, ab326932 was shown to bind specifically to DNA Polymerase theta. Target of interest was observed at 290 kDa 160 – 290kDa in wild-type HCT 116 cell lysates (lane 1) with no signal observed at this size in DNA Polymerase theta knockout cell line (lane 2) (lane 2, knockout cell line ab286468).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-DNA Polymerase theta antibody [EPR28988-1] (<a href='/en-us/products/primary-antibodies/dna-polymerase-theta-antibody-epr28988-1-ab326932'>ab326932</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 80 µg

Lane 2:

DNA Polymerase theta knockout HCT 116 whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 160-290 kDa,124 kDa

false

Exposure time: 180s

Western blot - Anti-DNA Polymerase theta antibody [EPR28988-1] - BSA and Azide free (AB326948)
  • WB

Lab

Western blot - Anti-DNA Polymerase theta antibody [EPR28988-1] - BSA and Azide free (AB326948)

This data was developed using ab326932, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-DNA Polymerase theta antibody [EPR28988-1] (<a href='/en-us/products/primary-antibodies/dna-polymerase-theta-antibody-epr28988-1-ab326932'>ab326932</a>) at 1/1000 dilution

Lane 1:

BT-549 (human Breast, Mammary gland epithelial) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

BT-549 transfected with siRNA specifically targeting DNA Polymerase theta whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 160-290 kDa,124 kDa

true

Exposure time: 180s

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR28988-1

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

We were unable to detect POLQ in tissue lysate samples.

Reactivity data

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Product details

ab326948 is the carrier-free version of ab326932

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

Low-fidelity DNA polymerase with a helicase activity that promotes microhomology-mediated end-joining (MMEJ), an alternative non-homologous end-joining (NHEJ) machinery required to repair double-strand breaks in DNA during mitosis (PubMed : 14576298, PubMed : 18503084, PubMed : 24648516, PubMed : 25642963, PubMed : 25643323, PubMed : 25775267, PubMed : 26636256, PubMed : 27311885, PubMed : 27591252, PubMed : 30655289, PubMed : 31562312, PubMed : 32873648, PubMed : 34140467, PubMed : 34179826, PubMed : 36455556, PubMed : 37440612, PubMed : 37674080). MMEJ is an error-prone repair pathway that produces deletions of sequences from the strand being repaired and promotes genomic rearrangements, such as telomere fusions, some of them leading to cellular transformation (PubMed : 25642963, PubMed : 25643323, PubMed : 25775267, PubMed : 27311885, PubMed : 27591252, PubMed : 31562312, PubMed : 32873648). MMEJ is required during mitosis to repair persistent double-strand breaks that originate in S-phase (PubMed : 37440612, PubMed : 37674080). Although error-prone, MMEJ protects against chromosomal instability and tumorigenesis (By similarity). The polymerase acts by binding directly the 2 ends of resected double-strand breaks, allowing microhomologous sequences in the overhangs to form base pairs (PubMed : 25643323, PubMed : 25775267, PubMed : 27311885, PubMed : 27591252). It then extends each strand from the base-paired region using the opposing overhang as a template (PubMed : 25643323, PubMed : 25775267, PubMed : 27311885, PubMed : 27591252). Requires partially resected DNA containing 2 to 6 base pairs of microhomology to perform MMEJ (PubMed : 25643323, PubMed : 25775267, PubMed : 27311885, PubMed : 27591252). The polymerase lacks proofreading activity and is highly promiscuous : unlike most polymerases, promotes extension of ssDNA and partial ssDNA (pssDNA) substrates (PubMed : 18503084, PubMed : 21050863, PubMed : 22135286). When the ends of a break do not contain terminal microhomology must identify embedded complementary sequences through a scanning step (PubMed : 32234782). Also acts as a DNA helicase, promoting dissociation of the replication protein A complex (RPA/RP-A), composed of RPA1, RPA2 and RPA3, from resected double-strand breaks to allow their annealing and subsequent joining by MMEJ (PubMed : 36455556). Removal of RPA/RP-A complex proteins prevents RAD51 accumulation at resected ends, thereby inhibiting homology-recombination repair (HR) pathway (PubMed : 25642963, PubMed : 28695890). Also shows RNA-directed DNA polymerase activity to mediate DNA repair in vitro; however this activity needs additional evidence in vivo (PubMed : 34117057). May also have lyase activity (PubMed : 19188258). Involved in somatic hypermutation of immunoglobulin genes, a process that requires the activity of DNA polymerases to ultimately introduce mutations at both A/T and C/G base pairs (By similarity). POLQ-mediated end joining activity is involved in random integration of exogenous DNA hampers (PubMed : 28695890).
See full target information POLQ

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