Anti-ROCK-1 Antibody, clone 3K23 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1879 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1879, Anti-ROCK-1, clone 3K23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of tubules & glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1879, Anti-ROCK-1, clone 3K23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of tubules & glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (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. ZRB1879, Anti-ROCK-1, clone 3K23 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of tubules & glomeruli of human kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZRB1879, Anti-ROCK-1, clone 3K23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane, cytosol, Golgi, and cytoskeleton.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZRB1879, Anti-ROCK-1, clone 3K23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane, cytosol, Golgi, and cytoskeleton.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of MCF-7 cells was performed using a 1:100 dilution of Cat. No. ZRB1879, Anti-ROCK-1, clone 3K23 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the membrane, cytosol, Golgi, and cytoskeleton.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from C6 (Lane 1), NIH3T3 (Lane 2), and HeLa (Lane 3) cells were probed with Cat. No. ZRB1879, Anti-ROCK-1, clone 3K23 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 ROCK-1 (~170 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 μg/mL of ROCK-1 peptide (Run 2) or non-specific control peptide (Run 3) were analyzed by affinity binding assay using Cat. No. ZRB1879, Anti-ROCK-1, clone 3K23 ZooMAb® Rabbit Monoclonal. The binding of this antibody to ROCK-1 peptide displayed a KD of 1.0 x 10-8 whereas binding to non-specific control peptide displayed a KD of 2.5 x 10-3.
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 | 3K23, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | C-terminus |
| 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 158.18 kDa |
| observed mol wt ~170 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_005397  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | rat, mouse, human |
| species reactivity (predicted by homology) | monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q13464 |
| Application: | Quality Control Testing Evaluated by Western Blotting in NIH3T3 cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected ROCK-1 in NIH3T3 cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected ROCK-1 in C6 and HeLa cell lysate. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected ROCK-1 in MCF-7 cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected ROCK-1 in human kidney tissue sections. Affinity Binding Assay: A representative lot of this antibody bound ROCK-1 peptide with a KD of 1.0 x 10-8 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 |
| 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 18 amino acids from the C-terminal region of human Rho-associated protein kinase 1 (ROCK-1). |
| 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 3K23 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects Rho-associated protein kinase 1 (ROCK-1). It targets an epitope within 18 amino acids from the C-terminal region. |
| 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: | Rho-associated protein kinase 1 (UniProt: Q13464; also known as EC:2.7.11.1, Renal carcinoma antigen NY-REN-35, Rho-associated coiled-coil-containing protein kinase 1, Rho-associated coiled-coil-containing protein kinase I, ROCK-I, p160 ROCK-1, p160ROCK) is encoded by the ROCK1 gene (Gene ID: 6093) in human. Rho kinase (ROCK), a serine/threonine kinase, serves as a target protein for small GTP-binding protein Rho and an important mediator of numerous cellular functions, including focal adhesions, motility, smooth muscle contraction, and cytokinesis. Rho kinase is found in two forms, ROCK1 and ROCK2 that share about 65% sequence homology. ROCK-1 contains an N-terminally located protein kinase domain (aa 76-338), a coiled coiled region, a Rho-binding domain (RBD; aa 949-1015) and a C-terminal pleckstrin homology (PH) domain (aa 1118-1317). It is considered as a key regulator of the actin cytoskeleton and cell polarity. Rho/GTP is reported to interact with the C-terminal portion of the central coiled-coil domain and activate the kinase activity. The C-terminal region, including the PH domain, acts as an autoinhibitory domain (aa 1115-1354) that binds directly to the kinase interface and deletion of this part leads to a constitutive activation in vitro and in vivo. Abnormal activation of the Rho/ROCK pathway has been observed in several disorders of the central nervous system. Brain and spinal cord injuries are reported to activate ROCKs, thereby inhibiting neurite growth and sprouting. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Hartmann, S., et al. (2015). Front. Pharmacol. 6; 276; Maekawa, M., et al.(1999). Science. 285(5429); 895-898). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |