PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30] - BSA and Azide free
Catalog# HA750419
PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30] - BSA and Azide free
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WB
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IHC-P
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IF-Tissue
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human PDK1 aa 16-65 / 436.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, IF-Tissue, IP
Target Molecular Weight
Predicted band size: 49 kDa
Positive Control
HepG2 cell lysate, HEK-293 cell lysate, LNCaP cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse heart tissue lysate, Rat heart tissue lysate, mouse heart tissue, rat heart tissue.
Conjugation
unconjugated
Clone Number
JA67-30
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
Mitochondrial pyruvate dehydrogenase (PDH) catalyzes the oxidative decarboxylation of pyruvate and plays a central role in the regulation of homeostasis of carbohydrate fuels in mammals. PDH activity is controlled by a phosphorylation/dephosphorylation cycle, phosphorylation leading to inactivation and dephosphorylation leading to reactivation of PDH. The phosphorylation of PDH is catalyzed by pyruvate dehydrogenase kinase (PDK), the activity of which is stimulated by the products of PDH catalysis. PDK1 consists of alpha and beta subunits; the kinase activity resides in the alpha subunit. Three PDK isoenzymes have been identified in humans (PDK1, 2 and 3) and two have been identified in rodent (PDK1 and 2).
Background References
1. Du L et al. Overexpression of PIK3CA in murine head and neck epithelium drives tumor invasion and metastasis through PDK1 and enhanced TGF signaling. Oncogene 35:4641-52 (2016).
2. Lelliott CJ et al. Monoclonal antibody targeting of fibroblast growth factor receptor 1c ameliorates obesity and glucose intolerance via central mechanisms. PLoS One 9:e112109 (2014).
Sequence Similarity
Belongs to the PDK/BCKDK protein kinase family.
Tissue Specificity
Expressed predominantly in the heart. Detected at lower levels in liver, skeletal muscle and pancreas.
Post-translational Modification
Phosphorylated by constitutively activated ABL1, FGFR1, FLT3 and JAK2 (in vitro), and this may also occur in cancer cells that express constitutively activated ABL1, FGFR1, FLT3 and JAK2. Phosphorylation at Tyr-243 and Tyr-244 strongly increases kinase activity, while phosphorylation at Tyr-136 has a lesser effect.
Subcellular Location
Mitochondrion matrix.
Synonyms
[Pyruvate dehydrogenase [lipoamide]] kinase isozyme 1, mitochondrial antibody
HGNC:8809 antibody
Mitochondrial pyruvate dehydrogenase kinase isoenzyme 1 antibody
PDH kinase 1 antibody
Pdk1 antibody
PDK1_HUMAN antibody
Pyruvate dehydrogenase kinase isoform 1 antibody
Pyruvate dehydrogenase kinase, isoenzyme 1 antibody
Images
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This data was developed using ET1704-66, the same antibody clone in a different buffer formulation.
Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (ET1704-66) at 1/5,000 dilution.
Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: HEK-293 cell lysate (15 µg/Lane)
Lane 3: LNCaP cell lysate (15 µg/Lane)
Lane 4: NIH/3T3 cell lysate (15 µg/Lane)
Lane 5: PC-12 cell lysate (15 µg/Lane)
Lane 6: Mouse heart tissue lysate (30 µg/Lane)
Lane 7: Rat heart tissue lysate (30 µg/Lane)
Predicted band size: 49 kDa
Observed band size: 45 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-66) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
This data was developed using ET1704-66, the same antibody clone in a different buffer formulation.
Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (ET1704-66) at 1/1,000 dilution.
Lane 1: A549-WT cell lysate
Lane 2: A549-KD PDK1 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 49 kDa
Observed band size: 46 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-66) at 1/1,000 dilution was used in primary antibody diluent at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1704-66, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-PDK1 antibody (ET1704-66) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-66) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1704-66, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-PDK1 antibody (ET1704-66) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-66) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1704-66, the same antibody clone in a different buffer formulation.
PDK1 was immunoprecipitated from 0.2 mg mouse heart tissue lysate with ET1704-66 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1704-66 at 1/2,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: Mouse heart tissue lysate (input)
Lane 2: ET1704-66 IP in mouse heart tissue lysate
Lane 3: Rabbit IgG instead of ET1704-66 in mouse heart tissue lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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