TIGAR/C12orf5 Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-49534

Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all TIGAR/C12orf5 Primary Antibodies »

Product Specifications

Immunogen

A genomic peptide made to an internal region of the human TIGAR protein (within residues 50-200). [Swiss-Prot Q9NQ88]

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for TIGAR/C12orf5 Antibody - BSA Free

Western Blot: TIGAR/C12orf5 AntibodyBSA Free [NBP1-49534]

Western Blot: TIGAR/C12orf5 AntibodyBSA Free [NBP1-49534]

Western Blot: TIGAR/C12orf5 Antibody [NBP1-49534] - Analysis of TIGAR in HeLa whole cell extracts.
Immunohistochemistry: TIGAR/C12orf5 Antibody - BSA Free [NBP1-49534]

Immunohistochemistry: TIGAR/C12orf5 Antibody - BSA Free [NBP1-49534]

Immunohistochemistry: TIGAR/C12orf5 Antibody [NBP1-49534] - Analysis of TIGAR in mouse muscle.

Applications for TIGAR/C12orf5 Antibody - BSA Free

Application
Recommended Usage

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Western Blot

1:5000
Application Notes
This TIGAR antibody is useful for Immunohistochemistry-paraffin sections and Western blot. In Western blot bands are seen ~30 kDa, representing TIGAR, and ~47 kDa. At this time we cannot explain the band at 47 kDa. This antibody does not appear to work in ICC.

Reviewed Applications

Read 1 review rated 5 using NBP1-49534 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: TIGAR/C12orf5

TIGAR (TP53-induced glycolysis and apoptosis regulator), a member of phosphoglycerate mutase family, is a target gene of p53 and it regulates glucose metabolism in cancer cells by acting as a fructose-bisphosphatase hydrolyzing fructose-2,6-bisphosphate (Fru-2,6-P2 ) and Fru-1,6-P2 leading to decreased Fru-2,6-P2 levels with subsequent reduction in PFK1 (phosphofructosekinase-1) activity as well as negative regulation of glycolysis. TIGAR also reduces cellular ROS through its translocation to the mitochondria where it interacts with HK2 leading to enhanced HK2 activity, regulation of mitochondrial membrane potential, and decreased mitochondrial ROS which ultimately protects the cells from DNA damage-induced apoptosis. Moreover, TIGAR mediated negative regulation of glycolysis leads to redirection of glycolytic metabolic intermediates towards pentose phosphate pathway's oxidative branch that follows increased cellular NADPH production, ROS scavenging by GSH and thus a lower sensitivity of cells to oxidative stress-associated apoptosis, including that induced by p53. Because of TIGAR's anti-apoptotic/protective role, it has been proposed to implicate in cell-cycle arrest and DNA repair effector response in cells that suffer from repairable dose of genotoxic/cellular insult.

Long Name

TP53-induced Glycolysis and Apoptosis Regulator

Alternate Names

C12orf5

Entrez Gene IDs

57103 (Human); 319801 (Mouse)

Gene Symbol

TIGAR

UniProt

Q9NQ88 (Human), Q8BZA9 (Mouse)

Additional TIGAR/C12orf5 Products

Product Documents for TIGAR/C12orf5 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for TIGAR/C12orf5 Antibody - BSA Free

Manufactured by Genomic Antibody Technology™. GAT FAQs

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Customer Reviews for TIGAR/C12orf5 Antibody - BSA Free (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: HepG2 total lysate, Sample Amount: ~ 50 ug
    Species: Human
    Verified Customer | Posted 01/30/2012
    TIGAR/C12orf5 Antibody - BSA Free NBP1-49534

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Protocols

View specific protocols for TIGAR/C12orf5 Antibody - BSA Free (NBP1-49534):

TIGAR/C12orf5 Antibody:
Immunohistochemistry-paraffin embedded sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer pH 6.0 then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench top for 30 minutes.

Staining:
1. Wash sections in dH2O three times for 5 minutes each.
2. Wash section in wash buffer (1X PBS/0.1% Tween-20 (1X PBST)) for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1X PBST, 5% goat serum) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul primary antibody diluted in 1X PBST, 5% goat serum to each section. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated secondary antibody, diluted in 1X PBST, 5% goat serum. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Striptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in dH2O.
12. Counterstain sections in hematoxylin.
13. Wash sections in dH2O two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

TIGAR/C12orf5 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% NFDM + 1% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-TIGAR primary antibody (NBP1-49534) in blocking buffer and incubate 1 hour at room temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).

**Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.

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FAQs

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