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Rabbit Recombinant Monoclonal ATR phospho S428 antibody. Carrier free. Suitable for I-ELISA, WB, IHC-P and reacts with Synthetic peptide - Human, Human, Mouse, Rat samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (AB316926), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (AB316926), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (AB316926), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (AB316926), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (AB316926), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
I-ELISAWBIHC-PICC/IF
Human
Expected
Tested
Tested
Not recommended
Mouse
Expected
Tested
Tested
Not recommended
Rat
Expected
Expected
Tested
Not recommended
Synthetic peptide - Human
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Expected
Expected

Species
Human, Mouse, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Human, Mouse
Dilution info
-
Notes

-

Expected
Expected

Species
Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Mouse
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Rat
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat, Synthetic peptide - Human
Dilution info
-
Notes

-

Target data

Function

Serine/threonine protein kinase which activates checkpoint signaling upon genotoxic stresses such as ionizing radiation (IR), ultraviolet light (UV), or DNA replication stalling, thereby acting as a DNA damage sensor (PubMed:10597277, PubMed:10608806, PubMed:10859164, PubMed:11721054, PubMed:12791985, PubMed:12814551, PubMed:14657349, PubMed:14729973, PubMed:14742437, PubMed:15210935, PubMed:15496423, PubMed:16260606, PubMed:21144835, PubMed:21777809, PubMed:23273981, PubMed:25083873, PubMed:27723717, PubMed:27723720, PubMed:30139873, PubMed:33848395, PubMed:37788673, PubMed:37832547, PubMed:9427750, PubMed:9636169). Recognizes the substrate consensus sequence [ST]-Q (PubMed:10597277, PubMed:10608806, PubMed:10859164, PubMed:11721054, PubMed:12791985, PubMed:12814551, PubMed:14657349, PubMed:14729973, PubMed:14742437, PubMed:15210935, PubMed:15496423, PubMed:16260606, PubMed:21144835, PubMed:23273981, PubMed:27723717, PubMed:27723720, PubMed:33848395, PubMed:9427750, PubMed:9636169). Phosphorylates BRCA1, CHEK1, MCM2, RAD17, RBBP8, RPA2, SMC1 and p53/TP53, which collectively inhibit DNA replication and mitosis and promote DNA repair, recombination and apoptosis (PubMed:11114888, PubMed:11418864, PubMed:11865061, PubMed:21777809, PubMed:23273981, PubMed:25083873, PubMed:9925639). Phosphorylates 'Ser-139' of histone variant H2AX at sites of DNA damage, thereby regulating DNA damage response mechanism (PubMed:11673449). Required for FANCD2 ubiquitination (PubMed:15314022). Critical for maintenance of fragile site stability and efficient regulation of centrosome duplication (PubMed:12526805). Acts as a regulator of the S-G2 transition by restricting the activity of CDK1 during S-phase to prevent premature entry into G2 (PubMed:30139873). Acts as a regulator of the nuclear envelope integrity in response to DNA damage and stress (PubMed:25083873, PubMed:37788673, PubMed:37832547). Acts as a mechanical stress sensor at the nuclear envelope: relocalizes to the nuclear envelope in response to mechanical stress and mediates a checkpoint via phosphorylation of CHEK1 (PubMed:25083873). Also promotes nuclear envelope rupture in response to DNA damage by mediating phosphorylation of LMNA at 'Ser-282', leading to lamin disassembly (PubMed:37832547). Involved in the inflammatory response to genome instability and double-stranded DNA breaks: acts by localizing to micronuclei arising from genome instability and catalyzing phosphorylation of LMNA at 'Ser-395', priming LMNA for subsequent phosphorylation by CDK1 and micronuclei envelope rupture (PubMed:37788673). The rupture of micronuclear envelope triggers the cGAS-STING pathway thereby activating the type I interferon response and innate immunity (PubMed:37788673). Positively regulates the restart of stalled replication forks following activation by the KHDC3L-OOEP scaffold complex (By similarity).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal ATR phospho S428 antibody. Carrier free. Suitable for I-ELISA, WB, IHC-P and reacts with Synthetic peptide - Human, Human, Mouse, Rat samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
Clone number
EPR25772-3F09
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C

Notes

ab316926 is the carrier-free version of Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925.

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

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

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.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

ATR also known as Ataxia Telangiectasia and Rad3-related protein is a serine/threonine kinase with a molecular weight of approximately 301 kDa. This protein localizes mainly in the nucleus where it functions as an important component in the cellular response to DNA damage and replication stress. ATR detects DNA strand breaks and ssDNA coated with RPA and becomes activated to phosphorylate several downstream targets initiating the DNA damage response. High expression of ATR occurs in proliferative tissues emphasizing its role in cell cycle regulation.

Biological function summary

ATR plays an essential role in maintaining genomic stability. It is part of a larger protein complex that includes ATRIP (ATR-interacting protein) which helps in localizing ATR to sites of DNA damage. Once activated ATR phosphorylates various substrates including CHK1 a critical checkpoint kinase involved in cell cycle arrest during DNA repair processes. The ability of ATR to coordinate with these proteins helps cells manage DNA damage effectively and prevent genomic instability.

Pathways

ATR functions centrally in the DNA damage response and repair mechanisms particularly the ATR-Chk1 pathway. This pathway interacts closely with the ATM (Ataxia Telangiectasia Mutated) pathway which also responds to DNA damage but usually to double-strand breaks. ATR primarily acts in response to replication stress and its activation leads to the arrest of the cell cycle allowing DNA repair to occur. This cooperation between ATR and ATM highlights their complementary roles in safeguarding genomic integrity under stress.

Associated diseases and disorders

ATR mutations and dysregulation have strong associations with cancer and Seckel syndrome. In the context of cancer ATR often works in concert with ATM to manage DNA repair and cancer cells frequently overexpress ATR to cope with high levels of replication stress. This makes ATR a potential target for cancer therapy where its inhibition could sensitize tumor cells to chemotherapy. In Seckel syndrome ATR mutations result in developmental anomalies showcasing the important role ATR plays in cellular replication and repair processes.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling ATR (phospho S428) with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining in human spleen without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.

    The section was incubated with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling ATR (phospho S428) with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining in rat spleen without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.

    The section was incubated with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Mouse Burkitt's lymphoma tissue labeling ATR (phospho S428) with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining in mouse Burkitt's lymphoma without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.

    The section was incubated with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling ATR (phospho S428) with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining in mouse spleen without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.

    The section was incubated with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling ATR (phospho S428) with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Positive staining in human ovarian cancer without alkaline phosphatase treatment; No signal was detected when tissues were treated with alkaline phosphatase.

    The section was incubated with Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 for 30 mins at room temperature.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

    Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

  • Indirect ELISA - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Indirect ELISA - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    Indirect ELISA analysis of Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1:2500 dilution.

    Antigen: Human ATR (phospho S428) peptide a,Human ATR (phospho S435) peptide b,Human ATR (phospho S428+S435) peptide c,Human ATR non-phospho peptide.

    Antigen concentration: 1000 ng/ml

  • Western blot - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Western blot - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

    In Western blot, Anti-ATR antibody - Total protein control staining at 1/1000 dilution.

    All lanes: Western blot - Anti-ATR (phospho S428) antibody [EPR25772-3F09] (Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925) at 1/1000 dilution

    Lane 1: Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate (untreated membrane) at 20 µg with NFDM/TBST

    Lane 2: RAW264.7 treated with 100J/m2 UV, then recovery for 4 hours, whole cell lysate (untreated membrane) at 20 µg with NFDM/TBST

    Lane 3: RAW264.7 treated with 100J/m2 UV, then recovery for 4 hours, whole cell lysate (Lambda phosphatase treated membrane) at 20 µg with NFDM/TBST

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 290 kDa, 124 kDa

    Exposure time: 180s

  • Western blot - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926), expandable thumbnail

    Western blot - Anti-ATR (phospho S428) antibody [EPR25772-3F09] - BSA and Azide free (ab316926)

    This data was developed using Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925, the same antibody clone in a different buffer formulation.

    The identity of the bands between 100 kDa and 130 kDa are unknown.

    In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] ab129002) staining at 1/10000 dilution.

    In Western blot, Anti-ATR antibody - Total protein control staining at 1/1000 dilution.

    All lanes: Western blot - Anti-ATR (phospho S428) antibody [EPR25772-3F09] (Anti-ATR (phospho S428) antibody [EPR25772-3F09] ab316925) at 1/1000 dilution

    Lane 1: Untreated HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg with NFDM/TBST

    Lane 2: HT-29 treated with 100mJ/cm2 UV, then recovery for 30 minutes, whole cell lysate (untreated membrane) at 20 µg with NFDM/TBST

    Lane 3: HT-29 treated with 100mJ/cm2 UV, then recovery for 30 minutes, whole cell lysate (Lambda phosphatase treated membrane) at 20 µg with NFDM/TBST

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Observed band size: 290 kDa, 124 kDa

    Exposure time: 180s

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Product protocols

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

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