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OGG1 Antibody - BSA Free

Catalog # NB100-106 | Novus Biologicals a Bio-Techne Brand

Key Product Details

Species Reactivity

Human, Mouse, Rat, Porcine, Primate, Rabbit

Applications

ELISA, Flow (Intracellular), Flow Cytometry, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Proximity Ligation Assay, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for OGG1 Antibody - BSA Free

Immunogen

A peptide derived from human Ogg1 (within amino acids 1-100). [UniProt# O15527]

Reactivity Notes

Rabbit reactivity reported in scientific literature (PMID: 16651612). Use in Porcine reported in scientific literature (PMID:28846530).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

39 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for OGG1 Antibody - BSA Free

Immunohistochemistry-Paraffin: OGG1 Antibody [NB100-106] - Analysis of OGG1 in human skin tissues. Image courtesy of product review by Arash Javeri.
Western Blot: OGG1 Antibody [NB100-106] - - Analysis of Ogg1 expression in Jurkat whole cell lysate.
Immunohistochemistry: OGG1 Antibody [NB100-106] - Figure 5 IHC-P analysis of hOGG1 protein expression in serous ovarian cancer: (A) Normal ovarian tissue treated with anti-hOGG1 antibody, biotinylated anti-rabbit IgG secondary antibody, and avidin-biotin-peroxidase complex, and counterstained with p-dimethylaminobenzaldehyde reagent (magnification 100 x); (B) serous cystadenoma with positive staining (magnification 200 x); (C) LG-SOC tissue with moderate positive staining (magnification 200 x); and (D) HG-SOC with negative staining (magnification 200 x). Black arrow pointed at the immunostained epithelial cells. Image collected and cropped by CiteAb from the following publication (http://ovarianresearch.biomedcentral.com/articles/10.1186/1757-2215-6-74), licensed under a CC-BY license.

Applications for OGG1 Antibody - BSA Free

Application
Recommended Usage

ELISA

reported in scientific literature (PMID 19506022)

Flow Cytometry

reported by customer review

Immunoblotting

reported in multiple pieces of scientific literature

Immunocytochemistry/ Immunofluorescence

1:100 - 1:200

Immunohistochemistry-Frozen

1:25-1:100

Immunoprecipitation

1:10-1:500. Use reported in scientific literature (PMID 20956573)

Proximity Ligation Assay

reported in scientific literature (PMID 35883652)

Western Blot

1:500-1:1000
Application Notes
Use in Immunoprecipitation has been reported in the literature Co-IP (PMID: 20956573). In WB, it recognizes a band at ~39 kDa, representing Ogg1. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 6 reviews rated 4 using NB100-106 in the following applications:

Published Applications

Read 128 publications using NB100-106 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: OGG1

8-hydroxyguanine, a form of oxidative DNA damage induced by free radicals, causes G:C to T:A transversion. In E. coli, three DNA repair enzymes exist to prevent the mutagenic effects of 8-hydroxyguanine. One of these enzymes, MutM, was found to have a functional yeast (yOgg1) and human (hOgg1) homologue. hOgg1 proteins efficiently release the 8-hydroxyguanine opposite the pyrimidine from DNA and cleave the AP site in a manner similar to bacterial and yeast enzymes. Genetic backgrounds in control of the repair of damaged DNA are involved in the susceptibility to cancer development. The hOgg1 gene has been mapped to region 3p26.2, a region showing loss of heterozygosity (LOH) in a variety of cancers. In particular, 3p25-p26 is a common LOH region in lung cancer. Recent work has demonstrated that Ogg plays an important role in CAG expansion, a characteristic of several neurodegenerative diseases. Ogg appears to be responsible for progressive expansion of poly-Q tracts in response to oxidative damage. Thus, Ogg provides a direct link between DNA damage and toxicity in neurons.

Long Name

8-oxoGuanine DNA Glycosylase

Alternate Names

8-hydroxyguanine DNA glycosylase, 8-oxoguanine DNA glycosylase, AP lyase, DNA-apurinic or apyrimidinic site lyase, HOGG1, MMH, MUTMOGH1HMMH, N-glycosylase/DNA lyase

Entrez Gene IDs

4968 (Human); 81528 (Rat)

Gene Symbol

OGG1

UniProt

Product Documents for OGG1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for OGG1 Antibody - BSA Free

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.

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Protocols

View specific protocols for OGG1 Antibody - BSA Free (NB100-106):

[[URL:https://www.novusbio.com/products/ogg1-antibody_nb100-106]][[Caption:OG… Antibody]]
Immunocytochemistry Protocol

Culture cells to appropriate density on suitable glass coverslips in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 5-10 minutes.
2. Remove the formalin and add 0.5% Triton-X 100 in TBS to permeabilize the cells. Incubate for 5-10 minutes.
3. Remove the permeabilization buffer and add wash buffer (i.e. PBS or PBS with 0.1% Tween-20). Be sure to not let the specimen dry out. Gently wash three times for 10 minutes.
4. Alternatively, cells can be fixed with -20C methanol for 10 min at room temperature. Remove the methanol and rehydrate in PBS for 10 min before proceeding.
5. To block nonspecific antibody binding incubate in 10% normal goat serum for 1 hour at room temperature.
6. Add primary antibody at appropriate dilution and incubate at room temperature for 1 hour or at 4 degrees C overnight.
7. Remove primary antibody and replace with wash buffer. Gently wash three times for 10 minutes.
8. Add secondary antibody at the appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove antibody and replace with wash buffer. Gently wash three times for 10 minutes.
10. Nuclei can be staining with 4',6' diamino phenylindole (DAPI) at 0.1 ug/ml, or coverslips can be directly mounted in media containing DAPI.
11. Cells can now be viewed with a fluorescence microscope.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow proper laboratory procedures for the disposal of formalin.
[[URL:https://www.novusbio.com/products/ogg1-antibody_nb100-106]][[Caption:OG… Antibody]]
Immunohistochemistry - FFPE sections

I. Deparaffinization:
A. Treat slides with Xylene: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.
B. Treat slides with 100% Reagent Alcohol: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.

II. Quench Endogenous Peroxidase:
A. Place slides in peroxidase quenching solution: 15-30 minutes.
To Prepare 200 ml of Quenching Solution:
Add 3 ml of 30% Hydrogen Peroxide to 200 ml of Methanol.
Use within 4 hours of preparation

B. Place slides in distilled water: 2 changes for 2 minutes each.

III. Retrieve Epitopes:
A. Preheat Citrate Buffer. Place 200 ml of Citrate Buffer Working Solution into container, cover and place into steamer. Heat to 90-96 degrees Celcius.
B. Place rack of slides into hot Citrate Buffer for 20 minutes. Cover.
C. Carefully remove container with slides from steamer and cool on bench, uncovered, for 20 minutes.
D. Slowly add distilled water to further cool for 5 minutes.
E. Rinse slides with distilled water. 2 changes for 2 minutes each.

IV. Immunostaining Procedure:
A. Remove each slide from rack and circle tissue section with a hydrophobic barrier pen (e.g. Liquid Blocker-Super Pap Pen).
B. Flood slide with Wash Solution. Do not allow tissue sections to dry for the rest of the procedure.
C. Drain wash solution and apply 4 drops of Blocking Reagent to each slide and incubate for 15 minutes.
D. Drain Blocking Reagent (do not wash off the Blocking Reagent), apply 200 ul of primary antibody solution to each slide, and incubate for 1 hour.
E. Wash slides with Wash Solution: 3 changes for 5 minutes each.
F. Drain wash solution, apply 4 drops of Secondary antibody to each slide and incubate for 1 hour.
G. Wash slides with Wash Solution: 3 changes for 5 minutes each.
H. Drain wash solution, apply 4 drops of DAB Substrate to each slide and develop for 5-10 minutes. Check development with microscope.
I. Wash slides with Wash Solution: 3 changes for 5 minutes each.
J. Drain wash solution, apply 4 drops of Hematoxylin to each slide and stain for 1-3 minutes. Increase time if darker counterstaining is desired.
K. Wash slides with Wash Solution: 2-3 changes for 2 minutes each.
L. Drain wash solution and apply 4 drops of Bluing Solution to each slide for 1-2 minutes.
M. Rinse slides in distilled water.
N. Soak slides in 70% reagent alcohol: 3 minutes with intermittent agitation.
O. Soak slides in 95% reagent alcohol: 2 changes for 3 minutes each with intermittent agitation.
P. Soak slides in 100% reagent alcohol: 3 changes for 3 minutes each with intermittent agitation. Drain slides for 10 seconds between each change.
Q. Soak slides in Xylene: 3 changes for 3 minutes each with intermittent agitation. Drain slides for 10 seconds between each change.
R. Apply 2-3 drops of non-aqueous mounting media to each slide and mount coverslip.
S. Lay slides on a flat surface to dry prior to viewing under microscope.

NOTES:
Use treated slides (e.g. HistoBond) to assure adherence of FFPE sections to slide.
Prior to deparaffinization, heat slides overnight in a 60 degrees Celcius oven.
All steps in which Xylene is used should be performed in a fume hood.
For Epitope Retrieval, a microwave or pressure cooker may be substituted for the steamer method. Adjust times as necessary depending on conditions.
For the initial IHC run with a new primary antibody, test tissues with and without Epitope Retrieval. In some instances, Epitope Retrieval may not be necessary.
200 ul is the recommended maximum volume to apply to a slide for full coverage. Using more than 200 ul may allow solutions to wick off the slide and create drying artifacts. For small tissue sections less than 200 ul may be used.
5 minutes of development with DAB Substrate should be sufficient. Do not develop for more than 10 minutes. If 5 minutes of development causes background staining, further dilution of the primary antibody may be necessary.
Hematoxylin should produce a light nuclear counterstain so as not to obscure the DAB staining. Counterstain for 1-1 1/2 minutes for nuclear antigens. Counterstain for 2-3 minutes for cytoplasmic and membranous antigens. If darker counterstaining is desired increase time (up to 10 minutes).
[[URL:https://www.novusbio.com/products/ogg1-antibody_nb100-106]][[Caption:OG… Antibody]]
Western Blot Procedure

1. Run 50 ug of protein on a 4-20% Tris-glycine mini-gel at 125V for 90 minutes.
2. Equilibrate gel, nitrocellulose membrane, Whatman paper, and blotting pads in transfer buffer for 15 minutes.
3. Transfer protein to the membrane at 25V for 90 minutes.
4. Allow membrane to air-dry.
5. Block membrane with 1XPBS/3% BSA for 1 hour at room temperature (23-27 degrees C).
6. Wash membrane twice, for 5 minutes each, with 1XPBS/0.05% Tween-20 (PBST).
7. Incubate membrane with NB100-106 (anti-hOGG1), diluted in 1XPBS/1% BSA, for 1 hour at room temperature.
8. Wash membrane once for 15 minutes, then four times for 5 minutes each, with PBST.
9. Incubate membrane with goat anti-rabbit IgG-HRP, diluted in 1XPBS/1% BSA, for 1 hour at room temperature.
10. Wash membrane once for 15 minutes, then four times for 5 minutes each, with PBST.
11. Detect cross-reacting proteins using Renaissance Chemiluminescence Reagent Plus kit from NEN Life Sciences.

FAQs for OGG1 Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: Would you please help confirm if NB100-106 would work with human OGG1 alpha?

    A: NB100-106 has been validated for use in human, primate and rat for the following applications: Western Blot, IHC-Paraffin and IHC-Frozen. The immunogen used for this antibody maps to the N-terminal region between amino acids 1-100. I believe OGG1 isoforms differ in the C-terminal regional and all have a conserved N-terminus which would indicate to me that our antibody would recognize all isoforms. To be sure, take a close look at the isoform of interest in and compare its sequence to OGG1. If the differences lie outside the 1-100 amino acid range then we are confident the antibody will work for OGG1 alpha.

  • Q: Is there a difference between Lot#J1 and Lot#L1 for Immunohistochemistry ? What does this product change for product specification between Lot#J1 and Lot#L1?

    A: No, there should not have any significant differences between these two lots, J-1 and L-1 applying to any application. The different is the production time for the lots, as they could be bled, purified, and/or gone through other manufacturing processes at the different time points. However sometimes some minor issues do happen, and we call it "lot-to-lot" variations. These minor changes could readily due to how close the breeding time respective to the boosting injections (the closer, the higher titers of IgG), the healthy conditions of the hosts (infection or others), etc. But the specificities nor applicability of the different lots should not be changed.

Showing  1 - 2 of 2 FAQs Showing All
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