LRP2 Antikörper (Low Density Lipoprotein Receptor-Related Protein 2) (AA 3411-3450) (Cy3)

Details for Product anti-LRP2 Antibody No. ABIN750991, Anbieter: Anmelden zum Anzeigen
Antigen
  • im:680590
  • lrp2
  • dbs
  • gp330
  • megalin
  • DBS
  • GP330
  • AI315343
  • AW536255
  • D230004K18Rik
  • Gp330
  • Megalin
  • low density lipoprotein receptor-related protein 2
  • low density lipoprotein receptor-related protein 2a
  • low-density lipoprotein receptor-related protein 2
  • low-density lipoprotein receptor-related protein 2-like
  • LRP2
  • lrp2a
  • LRP2.3
  • lrp2
  • LOC100183506
  • Lrp2
Epitop
AA 3411-3450
19
14
3
1
1
1
1
Reaktivität
Human, Maus, Ratte (Rattus)
63
19
19
Wirt
Kaninchen
57
4
1
Klonalität
Polyklonal
Konjugat
Dieser LRP2 Antikörper ist konjugiert mit Cy3
3
3
3
2
2
2
1
1
1
1
1
1
1
1
1
1
Applikation
Immunofluorescence (Paraffin-embedded Sections) (IF (p))
31
29
16
13
12
5
4
3
2
1
1
Optionen
Hersteller
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Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Chargennummer YEYY9
Validierte Anwendung Immunofluorescence
Positivkontrolle MCF7 cells
Negativkontrolle HeLa cells
Bewertung Strong signal was detected in positive control tissues and not in negative control tissues. Note that there was a small amount of signal generated in the negative control sample.
Primärantikörper
Isotypkontrolle
Sekundärantikörper
Kontrollen
  • Positive control: MCF7, LRP2 positive breast carcinoma cells (specimen known to contain the target protein) from ATCC.
  • Negative Control: HeLa, LRP2 negative cervical carcinoma cells (specimen known to not contain the target protein) from http://www.atcc.org/products/all/CCL-2.aspx.
  • Isotype Control: Normal rabbit serum IgG (cat # sc-2338) from SantaCruz Biotechnology.
  • Secondary antibody only control: MCF7, LRP2 positive breast carcinoma cells incubated with Goat anti-Rabbit CY3 secondary antibody only.
Protokoll
  • MCF7 and HeLa cell lines were grown directly on coverslips and fixed with 4% formaldehyde in PBS for 15 min at room temperature (RT).
  • Fixed cells were rinsed three times in PBS for 5 min each at RT.
  • Cells were blocked in 1X PBS/1% BSA/0.3% Triton™ X-100 to block unspecific binding of the antibodies for 60 min at RT.
  • Cells were incubated with fluorophore conjugated primary antibody diluted 1:250 and 1:100 in 1X PBS/1% BSA/0.3% Triton™ X-100 overnight at 4°C.
  • Cells were rinsed three times in PBS for 5 min each at RT.
  • Coverslips were mounted on slides with ProLong® Gold Antifade Reagent with DAPI.
  • Isotype control staining:
  • All the steps are done same as previously described until cells were blocked in 1X PBS/1% BSA/0.3% Triton™ X-100 to block unspecific binding of the antibodies for 60 min at RT. - Cells were incubated with 10% normal rabbit serum overnight at 4°C.
  • Cells were rinsed three times in PBS for 5 min each at RT.
  • Cells were incubated with goat anti rabbit CY3 conjugated secondary antibody for 60 min in dark at RT.
  • Cells were rinsed three times in PBS for 5 min each at RT.
  • Coverslips were mounted on slides with ProLong® Gold Antifade Reagent with DAPI. Secondary only staining:
  • All the steps are done same as previously described until cells were blocked in 1X PBS/1% BSA/0.3% Triton™ X-100 overnight at 4°C. - Cells were incubated with goat anti rabbit CY3 conjugated secondary antibody for 60 min in dark at RT.
  • Cells were rinsed three times in PBS for 5 min each at RT.
  • Coverslips were mounted on slides with ProLong® Gold Antifade Reagent with DAPI.
  • Stained cells were imaged with a Nikon C2+ confocal microscope.
Anmerkungen We did the first set of experiment with 1:250 dilution of the primary antibody but have observed lower level of expression of LRP2 in MCF7 (positive) cells (Figure 2). We repeated experiment with higher concentration of the primary antibody (1:100) and the expression level increased as shown in Figure 1. We have noticed lower level of expression of LRP2 in HeLa (negative) cells (Figure 3). We have used rabbit normal serum for isotype control to match the species with primary antibody (rabbit) and also we used bovine albumin serum (BSA) as blocking and antibody diluent buffer to reduce species cross reactivity.
Validierungsbilder
Immunofluorescence validation image for anti-Low Density Lipoprotein Receptor-Related Protein 2 (LRP2) (AA 3411-3450) antibody (Cy3) (ABIN750991) Figure 1: MCF7 cells stained at 1:250 with LRP2 (red) and a nuclear counterstain (blue).
Immunofluorescence validation image for anti-Low Density Lipoprotein Receptor-Related Protein 2 (LRP2) (AA 3411-3450) antibody (Cy3) (ABIN750991) Figure 2: MCF7 cells stained at 1:100 with LRP2 (red) and a nuclear counterstain (blue).
Immunofluorescence validation image for anti-Low Density Lipoprotein Receptor-Related Protein 2 (LRP2) (AA 3411-3450) antibody (Cy3) (ABIN750991) Figure 3: HeLa cells stained with LRP2 (red) and a nuclear counterstain (blue).
Immunofluorescence validation image for anti-Low Density Lipoprotein Receptor-Related Protein 2 (LRP2) (AA 3411-3450) antibody (Cy3) (ABIN750991) Figure 4: MCF7 cells stained isotype control (red) and a nuclear counterstain (blue).
Immunofluorescence validation image for anti-Low Density Lipoprotein Receptor-Related Protein 2 (LRP2) (AA 3411-3450) antibody (Cy3) (ABIN750991) Figure 1: MCF7 cells with only a secondary antibody (red) and a nuclear counterstain ...
Immunogen KLH conjugated synthetic peptide derived from human Megalin
Isotyp IgG
Reinigung Purified by Protein A.
Andere Bezeichnung Megalin (LRP2 Antibody Abstract)
Hintergrund

Acts together with cubilin to mediate HDL endocytosis (By similarity). May participate in regulation of parathyroid-hormone and para-thyroid-hormone-related protein release.

Subcellular location: Extracellular

Synonyms: DBS, GP33, Low-density lipoprotein receptor-related protein 2, LRP-2, Glycoprotein 33, Megalin, LRP2

Gen-ID 4036
UniProt P98164
Pathways Metabolism of Steroid Hormones and Vitamin D, Thyroid Hormone Synthesis, Hormone Transport
Applikationshinweise IF(IHC-P) 1:50-200
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Lagerung 4 °C
Informationen zur Lagerung Store at 4°C
Haltbarkeit 12 months
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