Anti-phospho-Chk2-Thr68 Antibody, clone 1B17 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1434 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in subsets of tumor cells of human colon cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in subsets of tumor cells of human colon cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in subsets of tumor cells of human colon cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in subsets of tumor cells of human colon cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in subsets of tumor cells of human colon cancer tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with UV light (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with UV light (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with UV light (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with UV light (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with UV light (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the nucleus.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from untreated HeLa cells (Lane 1) and HeLa cells treated with UV light (50 milliJoules with 1 h recovery) (Lane 2) were probed with Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-Chk2-Thr68 (~60 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or phospho-Chk2-Thr68 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal. The binding of this antibody to phospho-Chk2-Thr68 peptide displayed a KD of 2.5 x 10-7, whereas binding to non-specific control peptide displayed a KD of 1.4 x 10-4.
Peptide Inhibition Assay Enhanced Validation - Recombinant Antibody Technology Lysate from HeLa cells treated with UV light were probed with 1: 1,000 dilution of Cat. No. ZRB1434, Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal. Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-Chk2-Thr68 (~60 kDa). Lanes 1. HeLa cells without any peptide treatment 2. HeLa cells treated with phosphorylated Chk2-Thr68 3. HeLa cells treated with non-phosphorylated Chk2-Thr68 peptide
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 1B17, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminal |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 60.92 kDa |
| observed mol wt ~60 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_665861  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | O96017 |
| Application: | Quality Control Testing Evaluated by Western Blotting in lysate from HeLa cells treated with UV light. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Checkpoint kinase 2 (Chk2) in lysate from HeLa cells treated with UV light (50 milli Joules followed by recovery of 1 h), but not in untreated cells. Tested applications Peptide Inhibition Assay: Target band detection in lysate from UV light treated HeLa cells was prevented by pre-blocking of a representative lot with the phosphorylated Chk2-Thr68 peptide, but not the corresponding non-phosphopeptide. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected phospho-Chk2-Thr68 in human colon cancer tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-Chk2-Thr68 in HeLa cells treated with UV light. Affinity Binding Assay: A representative lot of this antibody bound phospho-Chk2-Thr68 peptide with a KD of 2.5 x 10-7 in an affinity binding assay. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-phospho-Chk2-Thr68, clone 1B17 ZooMAb® Rabbit Monoclonal, Cat. No. ZRB1434, detects phospho-Chk2-Thr68 and is used in Affinity Binding Assay, Immunocytochemistry, Immunohistochemistry, and Peptide Inhibition Assay, and Western Blotting. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 9 amino acids surrounding phosphothreonine 68 from human Checkpoint kinase 2 (Chk2). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb® formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1B17 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Checkpoint kinase 2 (Chk2) phosphorylated on Threonine 68. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Serine/threonine-protein kinase Chk2 (UniProt: O96017; also known as EC:2.7.11.1, CHK2 checkpoint homolog, Cds1 homolog, Hucds1, hCds1, Checkpoint kinase 2, Chk2) is encoded by the CHEK2 (also known as CDS1, CHK2, RAD53) gene (Gene ID: 11200) in human. Chk2 is a homodimeric nuclear serine/threonine protein kinase that is required for checkpoint-mediated cell cycle arrest, activation of DNA repair, and apoptosis in response to the presence of DNA double-strand breaks. Its protein kinase domain is localized to amino acids 220-486. It is highly expressed in testis, spleen, colon, and peripheral blood leukocytes and lower expression is found in other tissues. Following activation, Chk2 phosphorylates several effector molecules preferentially at the consensus sequence [L-X-R-X-X-S/T]. It is reported to regulate cell cycle checkpoint arrest through phosphorylation of Cdc25A, Cdc25B, and Cdc25C, inhibiting their activity, which can lead to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. Chk2 undergoes phosphorylation at serine 73 by polo-like kinase 3 in response to DNA damage, promoting phosphorylation at threonine 68 by ATM kinase. Phosphorylation at threonine 68 induces its homodimerization and allows autophosphorylation to proceed at threonine 383 and 387 in the T-loop/activation segment that makes it fully active. activation state . Thirteen different isoforms of Chk2 have been described that are produced by alternative splicing. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |