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NG2/CSPG4 Rabbit mAb (A24955)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse

Flow cytometry: 1X10^6 Jurkat cells (negative control, left) and SK-MEL-28 cells (right) were surface-stained with NG2/CSPG4 Rabbit mAb (A24955, 2 μg/mL, orange line) or ABflo® 647 Rabbit IgG isotype control (A22070, 5 μl/Test, blue line), followed by Alexa Fluor® 647 conjugated goat anti-rabbit pAb staining. Non-fluorescently stained cells were used as blank control (red line).

Flow cytometry: 1X10^6 SK-MEL-28 cells were surface-stained with ABflo® 647 Rabbit IgG isotype control (A22070, 5 μl/Test, left) or NG2/CSPG4 Rabbit mAb (A24955, 2 μg/mL, right).

ABclonal:Western blot - NG2/CSPG4 Rabbit mAb (A24955)

Western blot analysis of lysates from Mouse ovary using NG2/CSPG4 Rabbit mAb (A24955) at 1:1000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.

Western blot analysis of various lysates using NG2/CSPG4 Rabbit mAb (A24955)at 1:1000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Negative control (NC): HL-60
Exposure time: 10s.

ABclonal:Immunohistochemistry - NG2/CSPG4 Rabbit mAb (A24955)

Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using NG2/CSPG4 Rabbit mAb (A24955) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human liver tissue using NG2/CSPG4 Rabbit mAb (A24955) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse brain tissue using NG2/CSPG4 Rabbit mAb (A24955) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - NG2/CSPG4 Rabbit mAb (A24955)

Confocal imaging of SK-MEL-28 cells using NG2/CSPG4 Rabbit mAb (A24955, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Overview

Product name	NG2/CSPG4 Rabbit mAb
Catalog No.	A24955
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC64580

Background

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.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 1583-2224 of human NG2 (NP_001888.2).
Sequence	SLKGSQTLTVCPGSVQPLSSQTLRASSSAGTDPQLLLYRVVRGPQLGRLFHAQQDSTGEALVNFTQAEVYAGNILYEHEMPPEPFWEAHDTLELQLSSPPARDVAATLAVAVSFEAACPQRPSHLWKNKGLWVPEGQRARITVAALDASNLLASVPSPQRSEHDVLFQVTQFPSRGQLLVSEEPLHAGQPHFLQSQLAAGQLVYAHGGGGTQQDGFHFRAHLQGPAGASVAGPQTSEAFAITVRDVNERPPQPQASVPLRLTRGSRAPISRAQLSVVDPDSAPGEIEYEVQRAPHNGFLSLVGGGLGPVTRFTQADVDSGRLAFVANGSSVAGIFQLSMSDGASPPLPMSLAVDILPSAIEVQLRAPLEVPQALGRSSLSQQQLRVVSDREEPEAAYRLIQGPQYGHLLVGGRPTSAFSQFQIDQGEVVFAFTNFSSSHDHFRVLALARGVNASAVVNVTVRALLHVWAGGPWPQGATLRLDPTVLDAGELANRTGSVPRFRLLEGPRHGRVVRVPRARTEPGGSQLVEQFTQQDLEDGRLGLEVGRPEGRAPGPAGDSLTLELWAQGVPPAVASLDFATEPYNAARPYSVALLSVPEAARTEAGKPESSTPTGEPGPMASSPEPAVAKGGFLSFLEANMFS
Gene ID
Swiss Prot
Synonyms	NG2; MCSP; MCSPG; MSK16; CSPG4A; HMW-MAA; MEL-CSPG; NG2/CSPG4
Calculated MW	251kDa
Observed MW	330kDa

Applications

Reactivity	Human, Mouse
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:1000 - 1:4000 IHC-P 1:200 - 1:800 IF/ICC 1:200 - 1:800 FC 1:500 - 1:1000 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence Flow Cytometry
Positive samples	Mouse ovary, HeLa
Cellular location	Apical cell membrane, Cell projection, Extracellular side, Single-pass type I membrane protein, lamellipodium membrane.
Customer validation	IHC(Rattus norvegicus) IF(Rattus norvegicus)

Documents

Certificate of Compliance

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Research Area

NG2/CSPG4 Rabbit mAb images

Flow CytoMetry - NG2/CSPG4 Rabbit mAb (A24955)

Flow cytometry: 1X10^6 SK-MEL-28 cells were surface-stained with ABflo® 647 Rabbit IgG isotype control (A22070, 5 μl/Test, left) or NG2/CSPG4 Rabbit mAb (A24955, 2 μg/mL, right).

Western blot - NG2/CSPG4 Rabbit mAb (A24955)