Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸

Order now and get it on Tuesday March 02, 2021

Anti-SPARC antibody [EPR20121] (ab207743)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20121] to SPARC
Suitable for: WB, Flow Cyt, IP
Knockout validated
Reacts with: Human

Product name

Anti-SPARC antibody [EPR20121]
See all SPARC primary antibodies
Description

Rabbit monoclonal [EPR20121] to SPARC
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: SH-SY5Y and A-375 whole cell lysates; Human bone giant cell tumor lysate; Human serum and brain lysate. Flow Cyt: A-375 cells. IP: A-375 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20121
Isotype

IgG
Research areas

Western blot - Anti-SPARC antibody [EPR20121] (ab207743)

All lanes : Anti-SPARC antibody [EPR20121] (ab207743) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SPARC knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 35 kDa
Observed band size: 35 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab207743 observed at 35 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab207743 was shown to recognize SPARC in wild-type HAP1 cells as signal was lost at the expected MW in SPARC knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SPARC knockout samples were subjected to SDS-PAGE. Ab207743 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-SPARC antibody [EPR20121] (ab207743)

Flow cytometric analysis of 4% paraformaldehyde-fixed A-375 (Human malignant melanoma cell line) cells labeling SPARC with ab207743 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-SPARC antibody [EPR20121] (ab207743)

All lanes : Anti-SPARC antibody [EPR20121] (ab207743) at 1/1000 dilution

Lane 1 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 2 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lane 3 : Human bone giant cell tumor lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Predicted band size: 35 kDa
Observed band size: 43 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 30 seconds; Lane 2/3: 3 minutes.

The observed molecular weight is consistent with what has been described in the literature (PMID: 18442048).
Western blot - Anti-SPARC antibody [EPR20121] (ab207743)

All lanes : Anti-SPARC antibody [EPR20121] (ab207743) at 1/1000 dilution

Lane 1 : Human serum
Lane 2 : Human brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Predicted band size: 35 kDa
Observed band size: 43 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-SPARC antibody [EPR20121] (ab207743)

SPARC was immunoprecipitated from 0.35 mg of A-375 (Human malignant melanoma cell line) whole cell lysate with ab207743 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab207743 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: A-375 whole cell lysate, 10µg (Input).

Lane 2: ab207743 IP in A-375 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207743 in A-375 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.
Anti-SPARC antibody [EPR20121] (ab207743)

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