CSPG4 Recombinant Rabbit Monoclonal Antibody [PSH24-56]
cat.: HA724452
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: PSH24-56
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, NIH/3T3 (Mouse fibroblasts) cell lysate, Mouse brain tissue lysate, Mouse spinal cord tissue lysate, Mouse pancreas 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
  FC
  IP
1:5,000
1:200-1:5,000
1:1,000
1-2μg/sample
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
HA724452_1.jpg Fig1: Western blot analysis of CSPG4 on different lysates with Rabbit anti-CSPG4 antibody (HA724452) 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
Lane 6: NIH/3T3 (Mouse fibroblasts) cell lysate
Lane 7: N9 (Mouse microglial cells) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 3 seconds; ECL: K1801

MCF7 is a negative control (PMID: 20852124).
SK-BR-3 is a negative control (PMID: 20852124).
Negative expression of Cspg4 protein in N9 cells is consistent with the predicted expression pattern.

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724452, 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
HA724452_2.jpg Fig2: Western blot analysis of CSPG4 on different lysates with Rabbit anti-CSPG4 antibody (HA724452) 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: Mouse pancreas tissue lysate

Lysates/proteins at 30 µg/Lane.
Exposure time: 15 seconds; ECL: K1801
Mouse liver is a negative control (PMID: 23481707).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724452, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
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
HA724452_3.jpg Fig3: Western blot analysis of CSPG4 on different lysates with Rabbit anti-CSPG4 antibody (HA724452) at 1/5,000 dilution.

Lane 1: Rat brain tissue lysate
Lane 2: Rat liver tissue lysate
Lane 3: Rat pancreas tissue lysate

Lysates/proteins at 30 µg/Lane.
Exposure time: 2 minutes; ECL: K1801

Rat liver is a negative control (PMID: 23481707).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724452, 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
HA724452_4.jpg Fig4: 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: HA724452, 1/5,000, 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).
HA724452_5.jpg Fig5: Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblasts)

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: HA724452, 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).
HA724452_6.jpg Fig6: 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: HA724452 (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).
HA724452_7.jpg Fig7: Application: Flow Cytometry

Species: Mouse
Sample: NIH/3T3 (Mouse fibroblasts)

Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724452 (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.
HA724452_8.jpg Fig8: Immunoprecipitation (IP)

CSPG4 was immunoprecipitated in 0.2 mg SK-MEL-28 (Human malignant melanoma cell) cell lysate with HA724452 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724452 at 1/20,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: SK-MEL-28 cell lysate (input)
Lane 2: HA724452 IP in SK-MEL-28 cell lysate
Lane 3: Rabbit IgG instead of HA724452 in SK-MEL-28 cell lysate

Exposure time: 10 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724452, 1/20,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 251 kDa
Observed band size: 330 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.