CSPG4 Recombinant Rabbit Monoclonal Antibody [PSH24-57]
cat.: HA724453
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, IF-Tissue, FC
Clonality: Monoclonal
Clone number: PSH24-57
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 251 kDa
Isotype: IgG
Immunogen: Recombinant protein within mouse CSPG4 aa 151-600.
Positive control: A375 (Human amelanotic melanoma cells) cell lysate, SK-MEL-28 (Human malignant melanoma cells) cell lysate, HeLa (Human cervical adenocarcinoma cells) cell lysate, Mouse brain tissue lysate, Mouse spinal cord tissue lysate, Rat brain tissue lysate, Rat pancreas tissue lysate.
Subcellular location: Cell membrane, Apical cell membrane, Cell projection, lamellipodium membrane, Cell surface.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  IF-Tissue
  FC
1:5,000
1:200
1:1,000
1:200
1:1,000
Uniprot #: SwissProt: Q6UVK1 Human | Q8VHY0 Mouse | Q00657 Rat
Alternative names: 4732461B14Rik AN2 AN2 proteoglycan Chondroitin sulfate proteoglycan 4 (melanoma-associated) Chondroitin sulfate proteoglycan 4 Chondroitin sulfate proteoglycan NG2 Cspg4 Cspg4 chondroitin sulfate proteoglycan 4 CSPG4_HUMAN HMW-MAA HSN tumor-specific antigen Kiaa4232 MCSP MCSPG MEL-CSPG Melanoma chondroitin sulfate proteoglycan Melanoma-associated chondroitin sulfate proteoglycan MELCSPG MSK16 NG2
Images
HA724453_1.jpg Fig1: Western blot analysis of CSPG4 on different lysates with Rabbit anti-CSPG4 antibody (HA724453) at 1/5,000 dilution.

Lane 1: A375 (Human amelanotic melanoma cells) cell lysate
Lane 2: SK-MEL-28 (Human malignant melanoma cells) cell lysate
Lane 3: MCF7 (Human breast cancer cells) cell lysate
Lane 4: HeLa (Human cervical adenocarcinoma cells) cell lysate
Lane 5: SK-BR-3 (Human breast cancer cells) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 9 seconds; ECL: K1801

MCF7 is a negative control (PMID: 20852124).
SK-BR-3 is a negative control (PMID: 20852124).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724453, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 251 kDa
Observed band size: 280,330 kDa
HA724453_2.jpg Fig2: Western blot analysis of CSPG4 on different lysates with Rabbit anti-CSPG4 antibody (HA724453) at 1/5,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Mouse liver tissue lysate
Lane 3: Mouse spinal cord tissue lysate
Lane 4: Rat brain tissue lysate
Lane 5: Rat liver tissue lysate
Lane 6: Rat pancreas tissue lysate

Lysates/proteins at 30 µg/Lane.
Exposure time: 30 seconds; ECL: K1801

Liver is a negative control (PMID: 23481707).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724453, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 251 kDa
Observed band size: 330 kDa
HA724453_3.jpg Fig3: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: A375 (Human amelanotic melanoma cells)/SK-BR-3 (Human breast cancer cells)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724453, 1/200, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
Negative control: SK-Br-3 (PMID: 20852124).
HA724453_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Choroid plexus
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724453, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724453_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Brain
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724453, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724453_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Liver
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724453, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

Mouse liver tissue is a negative control (PMID: 23481707).
HA724453_7.jpg Fig7: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Brain
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724453, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA724453_8.jpg Fig8: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Liver
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724453, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

Rat liver tissue is a negative control (PMID: 23481707).
HA724453_9.jpg Fig9: Application: Immunofluorescence (IF-tissue)

Species: Mouse
Tissue: Brain
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724453, 1/200, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA724453_10.jpg Fig10: Application: Immunofluorescence (IF-tissue)

Species: Mouse
Tissue: Liver
Sample: Paraffin-embedded section

Liver is a negative control (PMID: 23481707).

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724453, 1/200, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA724453_11.jpg Fig11: Application: Immunofluorescence (IF-tissue)

Species: Rat
Tissue: Brain
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724453, 1/200, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA724453_12.jpg Fig12: Application: Immunofluorescence (IF-tissue)

Species: Rat
Tissue: Liver
Sample: Paraffin-embedded section

Liver is a negative control (PMID: 23481707).

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724453, 1/200, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA724453_13.jpg Fig13: Application: Flow Cytometry

Species: Human
Sample: A375 (Human amelanotic melanoma cells)/SK-BR-3 (Human breast cancer cells)

Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724453 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Negative control: SK-Br-3 (PMID: 20852124).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.