Anti-p-CREB (Ser133) Antibody, clone 1H13 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1118 - recombinant, expressed in HEK 293 cells
Synonym: Cyclic AMP-responsive element-binding protein 1
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human stomach 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. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in gastric mucosa of Human Stomach cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human stomach 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. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in gastric mucosa of Human Stomach cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human stomach 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. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Nuclear staining was observed in gastric mucosa of Human Stomach cancer tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with Phorbol-12-Myristate-13-Acetate (PMA) (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 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 Phorbol-12-Myristate-13-Acetate (PMA) (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 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 Phorbol-12-Myristate-13-Acetate (PMA)(C and D) was performed using a 1:100 dilution of Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 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 Phorbol-12-Myristate-13-Acetate (PMA)(C and D) was performed using a 1:100 dilution of Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 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 Phorbol-12-Myristate-13-Acetate (PMA) (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 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 Phorbol-12-Myristate-13-Acetate (PMA; 200 nM) (Lane 2) were probed with Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 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 p-CREB (Ser133) (~37 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-phosphorylated CREB control peptide (Run 1) or phospho-CREB serine 133 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 ZooMAb® Rabbit Monoclonal. The binding of this antibody to phospho-CREB serine 133 peptide displayed a KD of 1.0 x 10-12, whereas binding to non-phosphorylated CREB control peptide displayed a KD of 2.2 x 10-3.
Peptide Inhibition Assay Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa cells treated with Phorbol-12-Myristate-13-Acetate (PMA) were probed with Cat. No. ZRB1118, Anti-p-CREB (Ser133), clone 1H13 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. Immobilon® Signal Enhancer was used. Arrow indicates p-CREB (Ser133) (~37 kDa). Lanes 1: Incubated without any peptide (Control). 2: Incubated with phosphorylated CREB (Ser133) peptide. 3: Incubated with non- phosphorylated CREB (Ser133) 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 | 1H13, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Unknown |
| 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 36.39 kDa |
| observed mol wt ~37 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_604391.1  |
| 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) | mouse |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| peptide synthesis: suitable | |
| western blot: suitable | |
| UniProt accession no. | P16220 |
| Application: | Quality Control Testing Evaluated by Western Blotting in lysate from Phorbol-12-Myristate-13-A Western Blotting Analysis (WB): A 1:1,000 dilution of this antibody detected phospho-CREB (Ser133) in lysate from HeLa cells serum starved overnight and treated with Phorbol-12-Myristate-13-A Tested applications Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected phospho-CREB (Ser133) in human stomach cancer tissue sections. Affinity Binding Assay:: A representative lot of this antibody bound phospho-CREB (Ser133) peptide with a KD of 1.0 x 10-12 in an affinity binding assay. Peptide Inhibition Assay: Target band detection in HeLa cell lysate was prevented by preblocking of a representative lot with the phopho-CREB (Ser133) peptide, but not the corresponding non-phosphopeptide. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected p-CREB (Ser133) in HeLa cells treated with Phorbol-12-Myristate-13-A 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 14 amino acids surrounding phosphoserine 133 in isoform 2 of human CREB (CREB-A). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 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 1H13 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects CREB isoform 2 (CREB-A) phosphorylated on serine 133. |
| 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: | Cyclic AMP-responsive element-binding protein 1 (UniProt: P16220; also known as CREB-1, cAMP-responsive element-binding protein 1) is encoded by the CREB1 gene (Gene ID: 1385) in human. CREB is a phosphorylation-dependent transcription factor that stimulates transcription upon binding to the DNA cAMP response element (CRE; TGACGTCA) as a dimer. It is essential for the activation of a number of immediate early genes. It also plays a key role in promoting neuronal survival, differentiation, proliferation, and neurite outgrowth. Transcription activation is reported to be enhanced by the TORC coactivators. CREB has four main functional domains: a C-terminal basic/leucine zipper domain (bZIP; aa 269-327), the Kinase Inducible Domain at the N-terminal (KID; aa 87-146), and two glutamine-rich domains (Q1, and Q2). The Q1 and Q2 domains provide transactivation function in the absence of cAMP signaling and synergize with KID and bZIP to confer full activity to CREB in response to cAMP. CREB is phosphorylated on serine 133 by protein kinase A, which promotes the recruitment of the co-activator CREB-binding protein(CBP) and p300 that increases the transcription of CREB-dependent genes. CREB is also phosphorylated on serine 133 by mitogen- and stress-activated kinase 1/2 (MASK 1/2) in cells in response to the activation of MAPK signaling. Phosphorylation of CREB is sufficient to induce cellular gene expression in response to cAMP, however, additional promoter-bound factors are required for target gene activation by CREB in response to mitogen or stress signals, indicating the presence of components in the CREB pathway that discriminate between different types of inputs. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Wang, H., et al. (2018). Front. Mol. Neurosci. 11; 255; Naqvi, S., et al. (2014) Biochem. J. 458(3); 469-479; Mayr, B., and Montiminy, M. (2001). Nat. Rev. Mol. Cell Biol. 2(8); 599-609). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |