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Key Product Details

Species Reactivity

Human, Mouse, Rat, Porcine, Canine, Xenopus, Zebrafish

Applications

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

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

1 mg/ml

Product Summary for SLC31A1/CTR1 Antibody

Immunogen

A synthetic peptide derived from a C-terminal sequence of human SLC31A1/CTR1 [Uniprot: O15431]

Reactivity Notes

Canine reactivity reported in scientific literature (PMID: 29901089).

Localization

Cell Membrane

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for SLC31A1/CTR1 Antibody

Immunocytochemistry/Immunofluorescence: SLC31A1/CTR1 Antibody [NB100-402] - CTR1 antibody was tested in NIH-3T3 cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).
Immunohistochemistry: SLC31A1/CTR1 Antibody [NB100-402] - Panel 1: human CTR1 staining of breast cancer tissue. Panel 2: human CTR1-antigen competition in breast cancer tissue.
Immunohistochemistry: SLC31A1/CTR1 Antibody [NB100-402] - Left panel: Antibody staining of hCTR1 overexpressing cell line. Right panel: Antibody plus peptide competition staining.

Applications for SLC31A1/CTR1 Antibody

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry

1:250

Immunohistochemistry-Paraffin

1:250
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 33 publications using NB100-402 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Preservative

0.02% Sodium Azide

Concentration

1 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. Store at -20C long term. Avoid freeze-thaw cycles.

Background: SLC31A1

SLC31A1/CTR1 (solute carrier family 31 member 1/copper transporter 1) is a homotrimeric high-affinity, saturable copper importer implicated in dietary copper uptake. Homeostasis of copper is tightly controlled by inter-regulatory circuitry containing copper, Sp1, and CTR1, wherein Sp1 transcription factor acts as a copper sensor in modulating CTR1 expression which in turn controls cellular copper and Sp1 levels in a 3-way mutual regulatory loop. CTR1 mediated copper uptake is time, temperature, and pH dependent and specific for reduced form of Cu (I) and copper reduction has been proposed to be caused by Steap protein family or Dcytb protein (Cybrd1) both of which are reported to possess cupric reductase activity. The intracellular localization of CTR1 is variable in different cell lines: CTR1 is localized at the plasma membrane in HEK293, CaCo-2, and A2780 cells, whereas, in other cells such as HeLa, it is predominantly localized to vesicular compartments, and variability in trafficking rates between vesicular and plasma membrane compartments has been suggested as the cause of this differential localization. Post-translational O-linked glycosylation of CTR1 protects it against proteolytic cleavage of N terminal 17-kD fragment and the biological function of this cleaved variant or protease responsible is largely unknown. Selective knockout of Ctr1 in murine intestinal epithelial cells results in severe systemic copper deficiency, ataxia, and death prior to weaning.

Long Name

Solute Carrier Family 31 Member 1

Alternate Names

COPT1, CTR1, HCTR1

Gene Symbol

SLC31A1

Product Documents for SLC31A1/CTR1 Antibody

Certificate of Analysis

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

Product Specific Notices for SLC31A1/CTR1 Antibody

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 SLC31A1/CTR1 Antibody (NB100-402):

Immunocytochemistry Protocol

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

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.
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 at all times).

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.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three 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 manufacturer's instructions.

FAQs for SLC31A1/CTR1 Antibody

Showing  1 - 4 of 4 FAQs Showing All
  • Q: Could you provide the immunogen sequence of this antibody?

    A: The immunogen sequence for this product is considered proprietary, but I can tell you it falls within amino acids 160-190 of CTR1.

  • Q: We have bought and used the SLC31A1/CTR1 antibody (Novus Biologicals; catnr. NB100-402) successfully for our Western blot and IHC analysis. We want to publish this, but the reviewers urged us to do a control experiment using a corresponding blocking peptide. Do you have the peptide available by which we can block the specific binding of this anti-SLC31A1 antibody?

    A: Thank you for contacting us regarding your successful use of our product. In regards to your inquiry, I have done a search of our products, and unfortunately, we do not have the blocking peptide for that particular antibody (NB100-402). I apologize for any inconvenience this may cause you.

  • Q: Could you please tell me the targeted amino acids of the CTR1 antibody NB100-402?

    A: Unfortunately, we cannot disclose that information. However, since human SLC31A1 is a small protein (http://www.uniprot.org/uniprot/O15431#sequences), I anticipate that so called ".. A synthetic peptide derived from a C-terminal sequence .. " should be located within a.a. 140 - 190.

  • Q: What is the molecular weight of CTR1?  

    A: The theoretical molecular weight of CTR1 is 21kDa. The protein can run slightly higher in western blot due to PTMs.

Showing  1 - 4 of 4 FAQs Showing All
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