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Key Product Details

Species Reactivity

Human, Chicken

Applications

ELISA, Gel Super Shift Assays, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 1D2

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for SREBP2 Antibody (1D2) - BSA Free

Immunogen

Human SREBP2 protein (amino acids 48-403). [UniProt# Q12772]

Reactivity Notes

Chicken was reported in PMIDs 15333705 and 11907029.

Localization

Endoplasmic reticulum and Golgi apparatus membranes,cytoplasmic vesicle. Mature/processed form localized to nucleus.

Specificity

Recognizes the precursor and mature forms of human SREBP2.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for SREBP2 Antibody (1D2) - BSA Free

Western Blot: SREBP2 Antibody (1D2) [NBP1-54446] - Lane1: HEp-2 cells. Lane 2: MCF-7 cells. Lane 3: LNCaP cells. Lane 4: HT1080 cells. Lane 5: ZR75-1 cells.
Immunocytochemistry/Immunofluorescence: SREBP2 Antibody (1D2) [NBP1-54446] - SREBP2 (1D2) antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).

Applications for SREBP2 Antibody (1D2) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

10 ug/mL

Immunoprecipitation

10 ug/mg protein

Western Blot

2 - 5 ug/mL
Application Notes
In WB, this antibody is expected to detect the precursor as well as the mature forms of SREBP2. Use of this antibody in EMSA/GS (Gel Supershift Assay) and IP applications has been reported in PMIDs 15333705 and 15213220, respectively.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 20 publications using NBP1-54446 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G 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

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

Background: SREBP2

Sterol regulatory element-binding proteins (SREBPs) compose a family of transcriptional factors that regulate the expression of various genes required for the synthesis of phospholipids, fatty acids, and cholesterol. Out of the three isoforms present (SREBP-1a, SREBP-1c and SREBP-2), SREBP-2 has been implicated more directly in the synthesis of cholesterol during metabolism. Similar to SREBP-1a and SREBP-1c, SREBP-2 acts by first binding to sterol regulatory DNA sequences, and subsequently up-regulating the synthesis of enzymes involved in sterol biosynthesis. SREBP-2 is expressed in both the cytoplasm and nucleus on a subcellular level, depending on the state of the protein. It has been known to interact and form a complex with the SCAP protein at the ER, in turn inhibiting interaction between SCAP and SEC24B. This interaction prevents the cleavage of SREBP-2, which results in the synthesis of cholesterol and uptake of LDL despite high levels of present sterols. In recent research, new functions for the SREBP family have offered insight on the effect of lipid metabolism on various pathophysiological diseases (cancers, steatosis and immunity, PMID: 22154484).

Long Name

Sterol Regulatory Element Binding Transcription Factor 2

Alternate Names

bHLhd2, SREBF2

Entrez Gene IDs

6721 (Human)

Gene Symbol

SREBF2

UniProt

Product Documents for SREBP2 Antibody (1D2) - 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 SREBP2 Antibody (1D2) - 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 SREBP2 Antibody (1D2) - BSA Free (NBP1-54446):

[[URL:https://www.novusbio.com/products/srebp2-antibody-1d2_nbp1-54446]][[Cap… Antibody (1D2)]]
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.
[[URL:https://www.novusbio.com/products/srebp2-antibody-1d2_nbp1-54446]][[Cap… Antibody (1D2)]]
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 25 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute anti-SREBP2 (1D2) primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted 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 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 manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

FAQs for SREBP2 Antibody (1D2) - BSA Free

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  • Q: The species reactivity of this antibody is only human, right? I have done IHC staining for the tissue of SQ prostate cancer in nude mice using SREBP-2 antibody from other companies. But we can detect high background and non-specific staining. So we try to find a new antibody just against human SREBP-2.

    A: As long as the tissue you use is human there should not be any back ground noise. If you are using this Ab on mouse tissue you will have non specific binding as the host of this Ab is mouse.

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