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

Details for Product anti-LRP2 Antibody No. ABIN750991, Supplier: Log in to see
Antigen
  • im:680590
  • lrp2
  • dbs
  • gp330
  • megalin
  • DBS
  • GP330
  • AI315343
  • AW536255
  • D230004K18Rik
  • Gp330
  • Megalin
  • LDL 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
  • LDL receptor related protein 2 L homeolog
  • LRP2
  • lrp2a
  • LRP2.3
  • lrp2.L
  • LOC100183506
  • lrp2
  • Lrp2
Alternatives
anti-Human LRP2 antibody for ELISA
Epitope
AA 3411-3450
19
14
9
8
2
2
1
1
1
Reactivity
Human, Mouse (Murine), Rat (Rattus)
86
24
24
Host
Rabbit
71
13
1
Clonality
Polyclonal
Conjugate
This LRP2 antibody is conjugated to Cy3
5
5
5
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
Application
Immunofluorescence (Paraffin-embedded Sections) (IF (p))
43
41
20
15
12
9
4
3
2
2
2
2
1
1
Options
Supplier
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Supplier Product No.
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'Independent Validation' Badge
Lot Number YEYY9
Method validated Immunofluorescence
Positive Control MCF7 cells
Negative Control HeLa cells
Notes 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.
Primary Antibody
  • Antigen: Low Density Lipoprotein Receptor-Related Protein 2 (LRP2) antibody (Cy3)
  • Catalog number: ABIN750991
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  • Supplier catalog number: Log in to see
  • Lot number: YEYY9
Isotype Control Antibody
Secondary Antibody
  • Used only for the secondary only control, as primary antibody was directly conjugated to Cy3
  • Antibody: Cy3 Goat Anti-Rabbit IgG (H+L)
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Controls
  • 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.
Protocol
  • 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.
Experimental Notes 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.
Validation Images
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
Isotype IgG
Purification Purified by Protein A.
Plasmids, Primers & others Plasmids, Primers & others LRP2 products on genomics-online (e.g. as negative or positive controls)
Alternative Name Megalin (LRP2 Antibody Abstract)
Background

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

Gene ID 4036
UniProt P98164
Pathways Metabolism of Steroid Hormones and Vitamin D, Thyroid Hormone Synthesis, Hormone Transport
Application Notes IF(IHC-P) 1:50-200
Restrictions For Research Use only
Format Liquid
Concentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
Storage 4 °C
Storage Comment Store at 4°C
Expiry Date 12 months
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