WB
Relative expression (RE). Western blot analysis of NG2/CSPG4 on different lysates with Rabbit anti-NG2/CSPG4 antibody at 1/5,000 dilution. Lysates/proteins at 15 µg/Lane. Exposure time: 3 seconds; MCF7 is a negative control. SK-BR-3 is a negative control. 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: 1/5,000 in primary antibody dilution buffer, overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.WB
Relative expression (RE). Western blot analysis of NG2/CSPG4 on different lysates with Rabbit anti-NG2/CSPG4 antibody at 1/5,000 dilution. Lysates/proteins at 30 µg/Lane. Exposure time: 2 minutes; Rat liver is a negative control. Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: 1/5,000 in primary antibody dilution buffer, overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.ICC/IF
Relative expression (RE). 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: 1/5,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue). Negative control: SK-Br-3.ICC/IF
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: 1/200, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).FC
Relative expression (RE). 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: 1/1,000 (Red) compared with Rabbit IgG Isotype Control (Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (488), 15 minutes at room temperature. Negative control: SK-Br-3.FC
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: 1/1,000 (Red) compared with Rabbit IgG Isotype Control (Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (488), 15 minutes at room temperature.IP
Immunoprecipitation (IP). NG2/CSPG4 was immunoprecipitated in 0.2 mg SK-MEL-28 (Human malignant melanoma cell) cell lysate with NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody at 1/20,000 dilution. Anti-Rabbit IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SK-MEL-28 cell lysate (input) Lane 2: NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody IP in SK-MEL-28 cell lysate Lane 3: Rabbit IgG instead of NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody in SK-MEL-28 cell lysate Exposure time: 10 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody, 1/20,000 in primary antibody dilution buffer, 2 hours at room temperature.| Product Name | NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within mouse CSPG4 aa 151-600. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIPWB |
| WB:1:5000 ICC/IF:1:200-1:5000 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CSPG4 |
|---|---|
| Gene Synonyms | NG2 MCSP MCSPG MSK16 CSPG4A HMW-MAA MEL-CSPG |
| Gene Full Name | chondroitin sulfate proteoglycan 4 |
| Gene Summary | A human melanoma-associated chondroitin sulfate proteoglycan plays a role in stabilizing cell-substratum interactions during early events of melanoma cell spreading on endothelial basement membranes. CSPG4 represents an integral membrane chondroitin sulfate proteoglycan expressed by human malignant melanoma cells. [provided by RefSeq, Jul 2008] |
| Alternative Names | 4732461B14Rik antibody AN2 antibody AN2 proteoglycan antibody Chondroitin sulfate proteoglycan 4 (melanoma-associated) antibody Chondroitin sulfate proteoglycan 4 antibody Chondroitin sulfate proteoglycan NG2 antibody Cspg4 antibody Cspg4 chondroitin sulfate proteoglycan 4 antibody CSPG4_HUMAN antibody HMW-MAA antibody HSN tumor-specific antigen antibody Kiaa4232 antibody MCSP antibody MCSPG antibody MEL-CSPG antibody Melanoma chondroitin sulfate proteoglycan antibody Melanoma-associated chondroitin sulfate proteoglycan antibody MELCSPG antibody MSK16 antibody NG2 antibody |
| Molecular Weight(MW) | 251 kDa(Observed band size:330 kDa) |
| Cellular Localization | Cell membrane, Apical cell membrane, Cell projection, lamellipodium membrane, Cell surface. |
WB
Relative expression (RE). Western blot analysis of NG2/CSPG4 on different lysates with Rabbit anti-NG2/CSPG4 antibody at 1/5,000 dilution. Lysates/proteins at 15 µg/Lane. Exposure time: 3 seconds; MCF7 is a negative control. SK-BR-3 is a negative control. 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: 1/5,000 in primary antibody dilution buffer, overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.
WB
Relative expression (RE). Western blot analysis of NG2/CSPG4 on different lysates with Rabbit anti-NG2/CSPG4 antibody at 1/5,000 dilution. Lysates/proteins at 30 µg/Lane. Exposure time: 2 minutes; Rat liver is a negative control. Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: 1/5,000 in primary antibody dilution buffer, overnight at 4 °C Secondary antibody: Goat anti-Rabbit IgG-HRP, 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature.
ICC/IF
Relative expression (RE). 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: 1/5,000, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue). Negative control: SK-Br-3.
ICC/IF
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: 1/200, overnight at 4°C. Secondary antibody: Goat Anti-Rabbit IgG (488), 45 minutes at room temperature. Counterstain: Beta tubulin (red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
FC
Relative expression (RE). 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: 1/1,000 (Red) compared with Rabbit IgG Isotype Control (Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (488), 15 minutes at room temperature. Negative control: SK-Br-3.
FC
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: 1/1,000 (Red) compared with Rabbit IgG Isotype Control (Green), 15 minutes at room temperature. Secondary antibody: Goat Anti-Rabbit IgG (488), 15 minutes at room temperature.
IP
Immunoprecipitation (IP). NG2/CSPG4 was immunoprecipitated in 0.2 mg SK-MEL-28 (Human malignant melanoma cell) cell lysate with NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody at 1/20,000 dilution. Anti-Rabbit IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SK-MEL-28 cell lysate (input) Lane 2: NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody IP in SK-MEL-28 cell lysate Lane 3: Rabbit IgG instead of NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody in SK-MEL-28 cell lysate Exposure time: 10 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: NG2/CSPG4 Recombinant Rabbit Monoclonal Antibody, 1/20,000 in primary antibody dilution buffer, 2 hours at room temperature.| Application Notes | WB:1:5000 ICC/IF:1:200-1:5000 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
