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Western Blot (WB) (Western blot analysis of GAPDH expression in (1) Jurkat cell lysate; (2) A375 cell lysate; (3) Human hippocampus lysate; (4) Human fetal liver lysate; (5) COS-1 cell lysate; (6) Raw2647 cell lysate; (7) Mouse kidney lysate; (8) PC-12 cell lysate; (9) Rat brain lysate with GAPDH Antibody (MBS179033).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH monoclonal antibody (MBS179033) overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for GAPDH )

Rabbit GAPDH Monoclonal Antibody | anti-GAPDH antibody

Anti-GAPDH Rabbit Monoclonal Antibody, HRP Conjugated

Gene Names
GAPDH; G3PD; GAPD; HEL-S-162eP
Reactivity
Human, Mouse, Rat
Applications
Western Blot
Purity
Affinity-chromatography
Synonyms
GAPDH; Monoclonal Antibody; Anti-GAPDH Rabbit Monoclonal Antibody; HRP Conjugated; Glyceraldehyde-3-phosphate dehydrogenase; 1.2.1.12; Peptidyl-cysteine S-nitrosylase GAPDH; 2.6.99.-; GAPD; CDABP0047; OK/SW-cl.12; anti-GAPDH antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Monoclonal
Purity/Purification
Affinity-chromatography
Form/Format
Liquid
Sequence Length
293
Applicable Applications for anti-GAPDH antibody
Western Blot (WB)
Application Notes
WB: 1:5000-1:20000
Immunogen
A synthesized peptide derived from human GAPDH(HRP conjugated)
Subcellular Localization
Cytoplasm, cytosol. Nucleus. Cytoplasm, perinuclear region. Membrane. Cytoplasm, cytoskeleton. Translocates to the nucleus following S- nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
Contents
Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Western blot analysis of GAPDH expression in (1) Jurkat cell lysate; (2) A375 cell lysate; (3) Human hippocampus lysate; (4) Human fetal liver lysate; (5) COS-1 cell lysate; (6) Raw2647 cell lysate; (7) Mouse kidney lysate; (8) PC-12 cell lysate; (9) Rat brain lysate with GAPDH Antibody (MBS179033).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH monoclonal antibody (MBS179033) overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for GAPDH )

Western Blot (WB) (Western blot analysis of GAPDH expression in (1) Jurkat cell lysate; (2) A375 cell lysate; (3) Human hippocampus lysate; (4) Human fetal liver lysate; (5) COS-1 cell lysate; (6) Raw2647 cell lysate; (7) Mouse kidney lysate; (8) PC-12 cell lysate; (9) Rat brain lysate with GAPDH Antibody (MBS179033).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH monoclonal antibody (MBS179033) overnight at 4 degree C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for GAPDH )
Related Product Information for anti-GAPDH antibody
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D- glyceroyl phosphate. Component of the GAIT (gamma interferon- activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
36053 MW
NCBI Official Full Name
glyceraldehyde-3-phosphate dehydrogenase isoform 2
NCBI Official Synonym Full Names
glyceraldehyde-3-phosphate dehydrogenase
NCBI Official Symbol
GAPDH
NCBI Official Synonym Symbols
G3PD; GAPD; HEL-S-162eP
NCBI Protein Information
glyceraldehyde-3-phosphate dehydrogenase
UniProt Protein Name
Glyceraldehyde-3-phosphate dehydrogenase
UniProt Gene Name
GAPDH
UniProt Synonym Gene Names
GAPD; GAPDH

NCBI Description

This gene encodes a member of the glyceraldehyde-3-phosphate dehydrogenase protein family. The encoded protein has been identified as a moonlighting protein based on its ability to perform mechanistically distinct functions. The product of this gene catalyzes an important energy-yielding step in carbohydrate metabolism, the reversible oxidative phosphorylation of glyceraldehyde-3-phosphate in the presence of inorganic phosphate and nicotinamide adenine dinucleotide (NAD). The encoded protein has additionally been identified to have uracil DNA glycosylase activity in the nucleus. Also, this protein contains a peptide that has antimicrobial activity against E. coli, P. aeruginosa, and C. albicans. Studies of a similar protein in mouse have assigned a variety of additional functions including nitrosylation of nuclear proteins, the regulation of mRNA stability, and acting as a transferrin receptor on the cell surface of macrophage. Many pseudogenes similar to this locus are present in the human genome. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Nov 2014]

Uniprot Description

Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation.

Research Articles on GAPDH

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Product Notes

The GAPDH gapdh (Catalog #AAA179033) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-GAPDH Rabbit Monoclonal Antibody, HRP Conjugated reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's GAPDH can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: 1:5000-1:20000. Researchers should empirically determine the suitability of the GAPDH gapdh for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "GAPDH, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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