Anti-IRAK-1 Antibody, clone 3B12 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1975 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in lamina propria & subsets of glandular cells of human colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in lamina propria & subsets of glandular cells of human colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in lamina propria & subsets of glandular cells of human colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in lamina propria & subsets of glandular cells of human colon tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human colon (A and B) and Negative control (no primary) (C and D) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/nuclear staining was observed in lamina propria & subsets of glandular cells of human colon tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 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 cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 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 cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology HEK293 cell lysate was probed with Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 ZooMAb® Rabbit Monoclonal (1:250 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates IRAK-1 (~70 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or IRAK-1 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1975, Anti-IRAK-1, clone 3B12 ZooMAb® Rabbit Monoclonal. The binding of this antibody to IRAK-1 peptide displayed a KD of 1.0 x 10-12 whereas binding to non-specific control peptide displayed a KD of 2.0 x 10-5.
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 | 3B12, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Internal |
| 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 76.54 kDa |
| observed mol wt ~70 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_001560  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| 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. | P51617 |
| Application: | Quality Control Testing Evaluated by Immunohistochemistry (Paraffin) in human colon tissue sections. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution of this antibody detected IRAK-1 in human colon tissue sections. Tested applications Immunocytochemistry Analysis: A 1:1000 dilution from a representative lot detected IRAK-1 in K562 cells. Western Blotting Analysis: A 1:250 dilution from a representative lot detected IRAK-1 in HEK293 cell lysate. Affinity Binding Assay:: A representative lot of this antibody bound IRAK-1 with a KD of 1.0 x 10-12 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 24 amino acids from the internal region of human Interleukin-1 receptor-associated kinase 1 (IRAK-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 3B12 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects Interleukin-1 receptor-associated kinase 1 (IRAK-1). It targets an epitope within 24 amino acids from the internal 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: | Interleukin-1 receptor-associated kinase 1 (UniProt: P51617; also known as EC:2.7.11.1, IRAK-1) is encoded by the IRAK1 (also known as IRAK) gene (Gene ID: 3654) in human. The IRAK family is composed of IRAK-1, -2, and -4, which are expressed in a variety of human immune cell types and IRAK-M whose expression is largely limited to monocytes and macrophage. IRAK-1 is a ubiquitously expressed serine/threonine-protein kinase that play a central role in inflammatory responses by regulating the expression of various inflammatory genes in immune cells. These signals are critical for elimination of viruses, bacteria, and cancer cells, as well as for wound healing. IRAK-1 is involved in Toll-like receptor (TLR) and IL-1R signal transduction pathways. Is rapidly recruited by MYD88 to the receptor-signal transduction complex upon TLR activation. Following recruitment on the activated receptor complex, it is phosphorylated on Thr-209, probably by IRAK-4, resulting in a conformational change of the kinase domain (aa 212-521) that allows further phosphorylations to take place. Thr-387 phosphorylation in the activation loop is required to achieve full enzymatic activity. IRAK-1 signal transduction is considered to be important in the development and progression of cancer. Helicobacter pylori induced gastric inflammation is reported to up-regulate TLR2 and TLR5 expression, which in turn increases IRAK-1 phosphorylation and activation and NF- B activation. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Jain, A., et al. (2014). Front. Immunol. 5; 553; Kollewe, C., et al. (2004). J. Biol. Chem. 279(7); 5227-5236). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |