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Key Product Details

Species Reactivity

Mouse, Rat

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for TRPM2 Antibody - BSA Free

Immunogen

Synthetic peptide made to a C-terminal portion of the rat TRPM2 protein (within residues 1430-1508). [Swiss-Prot# Q5G856]

Localization

Cell membrane.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

181 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 TRPM2 Antibody - BSA Free

Western Blot: TRPM2 Antibody [NB110-81601] - Lane 1: CHO cells untransfected. Lane 2: CHO cell transfected with rat TRPM2 Lane 3: Mouse cortical neurons (cultured) Photo courtesy of Dr. Paco Herson, Oregon Health & Science University.
Immunocytochemistry/Immunofluorescence: TRPM2 Antibody [NB110-81601] - Neuro2a cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-TRPM2 at 5 ug/mL overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunocytochemistry/Immunofluorescence: TRPM2 Antibody [NB110-81601] - Staining of rat hippocampus using NB110-81601. TRPM2 is stained red, GFAP is stained green and nuclei staining is blue. Photo courtesy of Dr. Ji-Zhong Bai, The University of Auckland, New Zealand.

Applications for TRPM2 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:10 - 1:500

Immunohistochemistry

1:200

Immunohistochemistry-Paraffin

1:200

Western Blot

1:500 - 1:1000
Application Notes
In Western blot a band is seen at ~181 kDa. 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.

Published Applications

Read 12 publications using NB110-81601 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% 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

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

Background: TRPM2

TRP7 (transient receptor protein 7 or TRPC7) is the seventh identified member of mammalian TRPC channel family which includes nonselective cation channels that mediate changes in plasma membrane cation permeability induced by stimulation of PLC (phospholipase C, the cellular integrator that receives input from GPCRs/ RTK). Major cellular function of TRPC channels is to regulate Ca2+ influx across plasma membrane induced by stimulation of these receptors by hormones, neurotransmitters, and growth factors. Localized in the cell membrane and nuclear envelope as multi-pass membrane protein, TRPC7 interacts with with MX1, RNF24, PRKG1 etc. and PRKG1 mediated TRPC7 phosphorylation at Thr-15 site negatively regulates TRPC7's activity. TRP7 has been suggested to form a receptor-activated non-selective calcium permeant cation channel and is operated by a phosphatidylinositol second messenger system activated by RTKs or GPCRs. TRP7 is activated by DAG (diacylglycerol) and also upon intracellular calcium store depletion.

Alternate Names

EC 3.6.1.13, EREG1MGC133383, Estrogen-responsive element-associated gene 1 protein, KNP3LTrpC-2, Long transient receptor potential channel 2, LTrpC2, LTRPC2TRPC7transient receptor potential cation channel subfamily M member 2, NUDT9H, NUDT9L1, transient receptor potential cation channel, subfamily M, member 2, Transient receptor potential channel 7, TrpC7

Gene Symbol

TRPM2

Product Documents for TRPM2 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 TRPM2 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 TRPM2 Antibody - BSA Free (NB110-81601):

TRPM2 Antibody: https://www.novusbio.com/products/trpm2-antibody_nb110-81601
Immunocytochemistry Protocol

Culture cells to appropriate density 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 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
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 (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.
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