Anti-PDHA1 Antibody, clone 1B13 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1914 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney human retina (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. ZRB1914, Anti-PDHA1, clone 1B13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in tubules & glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney human retina (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. ZRB1914, Anti-PDHA1, clone 1B13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in tubules & glomeruli of human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney human retina (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. ZRB1914, Anti-PDHA1, clone 1B13 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in tubules & glomeruli of human kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A549 cells was performed using a 1:100 dilution of Cat. No. ZRB1914, Anti-PDHA1, clone 1B13 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 mitochondria.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A549 cells was performed using a 1:100 dilution of Cat. No. ZRB1914, Anti-PDHA1, clone 1B13 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 mitochondria.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A549 cells was performed using a 1:100 dilution of Cat. No. ZRB1914, Anti-PDHA1, clone 1B13 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 mitochondria.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from Rat liver tissue (Lane 1), Human kidney tissue (Lane 2), and NIH3T3 cells (Lane 3) were probed with Cat. No. ZRB1914, Anti-PDHA1, clone 1B13 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 PDHA1 (~40 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/ml of non-specific control peptide (Run 1) or PDHA1 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1914, Anti-PDHA1, clone 1B13 ZooMAb® Rabbit Monoclonal. The binding of this antibody to PDHA1 peptide displayed a KD of 2.6 x 10-7 whereas binding to non-specific control peptide displayed a KD of 2.0 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million A549 cells was performed using 1 μg of Cat. No. ZRB1914, Anti-PDHA1, clone 1B13 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG secondary antibody.
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 | 1B13, 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 half |
| 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 43.3 kDa |
| observed mol wt ~40 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_000275  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, mouse, rat |
| species reactivity (predicted by homology) | bovine, porcine, monkey |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P08559 |
| Application: | Quality Control Testing Evaluated by Western Blotting in Rat liver tissue lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected PDHA1 in Rat liver tissue lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected PDHA1 in lysates from human kidney tissue and NIH3T3 cells. Affinity Binding Assay: A representative lot of this antibody bound PDHA1 with a KD of 2.6 x 10-7 in an affinity binding assay. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected PDHA1 in human kidney tissue sections. Flow Cytometry Analysis: 1 μg from a representative lot detected PDHA1 in one million A549 cells. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected PDHA1 in A549 cells. 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 19 amino acids from the N-terminal half of human Pyruvate dehydrogenase E1a (PDHA1). |
| 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 1B13 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects PDHA1. It targets an epitope within 19 amino acids from the N-terminal half. |
| 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: | Pyruvate dehydrogenase E1 component subunit alpha, somatic form, mitochondrial (UniProt: P08559; also known as EC:1.2.4.1, PDHE1-A type I) is encoded by the PDHA1 (also known as PHE1A) gene (Gene ID: 5160) in human. The pyruvate dehydrogenase complex catalyzes the overall conversion of pyruvate to acetyl-CoA and CO2, and thereby links the glycolytic pathway to the tricarboxylic cycle. The pyruvate dehydrogenase complex (PDC) is a tightly organized arrangement of polypeptides and cofactors that work in a sequential manner in the multi-enzyme complex catalyzing a multi-step reaction in which pyruvate is converted into acetyl-CoA and CO2. The intermediate substrates and products remain bound to the complex, increasing overall efficiency. This complex contains three catalytic enzymes (E1, E2, E3), two regulatory enzymes, and a binding protein and requires cofactors TPP, lipoic acid, and FAD. E1 (PDHA1) is a ubiquitously distributed mitochondrial matrix protein that is synthesized with a mitochondrial transit peptide (aa 1-29), which is subsequently cleaved off to produce the mature active form. It is considered as a rate limiting step of PDC. Its activity is regulated by phosphatases that activate it and kinases that inhibit the enzyme activity. Phosphorylation at Ser-232, Ser-293 and Ser-300 by PDK family of kinases is known to inactivate the enzyme and phosphorylation at a single site is sufficient to cause this inactivation. However, dephosphorylation at all three sites is required for reactivation. E2 is responsible for transferring the hydroxyethyl group from TPP to an oxidized form of covalently bound lipoamide, and the resulting acetyl group is then transferred to free coenzyme A to form acetyl-CoA and reduced dihydrolipoamide-E2. E3 re-oxidizes the lipoyl group of dihydrolipoamide-E2 to form lipoamide-E2 and NADH. Mutations in PDHA1 gene are known to cause E1 deficiency that results in primary lactic acidosis in children. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |