ASM24

antibody from Cytoskeleton, Inc.

Targeting: SUMO3 SMT3A, SMT3H1

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

ASM24

Provider

Cytoskeleton, Inc.

Product name

Anti-SUMO-2/3

Antibody type

Monoclonal

Antigen

Full-length recombinant SUMO-2 protein (Uniprot: P61956) combined with a proprietary mix of peptides that include CQIRFRFDGQPINE.

Description

Anti-SUMO-2/3 antibody is a mouse monoclonal antibody. The antibody was raised against full-length recombinant SUMO-2 protein (Uniprot: P61956) combined with a proprietary mix of peptides that include CQIRFRFDGQPINE. The antibody has been shown to immunoprecipitate a wide range of SUMO-2/3 targeted proteins in a HeLa cell lysate (Fig. 1A). A linear epitope has not been identified and ASM24 appears to recognize a conformational epitope. Each Lot of antibody is quality controlled to provide a high batch to batch consistency. The Lot specific µg per tube can be found in the Lot specific COA documents at www.cytoskeleton.com. ASM24 is purified by Protein G affinity chromatography and is supplied as a lyophilized white powder. Each Lot of antibody is quality controlled to provide a high batch to batch consistency. The Lot specific µg per tube can be found in the Lot specific COA documents at www.cytoskeleton.com.

Host

Mouse

Conjugate

Unconjugated

Antigen sequence

Full-length recombinant SUMO-2 prot
ein (Uniprot: P61956) combined with
a proprietary mix of peptides that
include CQIRFRFDGQPINE.


Epitope

unidentified

Isotype

IgG

Antibody clone number

11G2

Vial size

2 x 200µl

Concentration

Specified in COA

Storage

Lyophilized 4 °C. Reconstituted -20 °C.

Handling

Shipped at room temperture. The lyophilized protein can be stored desiccated at 4 °C for 6 months. Store at -20 °C when reconstituted.

Immunocytochemistry

Supportive validation

Submitted by

Cytoskeleton, Inc. (provider)

Main image

Experimental details

HeLa cells were stained and visualized by widefield fluorescence microscopy as de-scribed in the IF method below. The cells were stained against β-tubulin (sheep anti-tubulin Ab, Cat# ATN02, green) and SUMO-2/3 (11G2, red). DNA was stained with DAPI. Cells in interphase were imaged with a Zeiss Axio Observer.Z1 microscope (1.4 NA 63X objective). PML nuclear bodies (nuclear dots) were visible in SUMO-2/3 staining as has been previously reported5. To see the full Immunofluorescence protocol, see the product datasheet.

Sample type

HeLa cells

Primary Ab dilution

1:500

Secondary Ab

fluorescently-labeled donkey anti-mouse

Secondary Ab dilution

1:500

Protocol

Protocol

Supportive validation

Submitted by

Cytoskeleton, Inc. (provider)

Main image

Experimental details

HeLa cells were stained and visualized by widefield fluorescence microscopy as de-scribed in the IF method below. The cells were stained against α/β-tubulin (sheep anti-tubulin Ab, Cat# ATN02, green) and SUMO-2/3 (11G2, red). DNA was stained with DAPI. Mitotic cells in metaphase were imaged with a Zeiss Axio Observer.Z1 microscope (1.4 NA 63X objective). The localization of SUMO 2/3-conjugates at chromosomes can be observed during mitosis as has been previously reported12. To see the full Immunofluorescence protocol, see the product datasheet.

Sample type

HeLa cells

Primary Ab dilution

1:500

Secondary Ab

fluorescently-labeled donkey anti-mouse

Secondary Ab dilution

1:500

Protocol

Protocol

Immunoprecipitation

Supportive validation

Submitted by

Cytoskeleton, Inc. (provider)

Main image

Experimental details

Denatured cell lysates were prepared from HS43, CT37 and KD S210 (HS43: Heat Shock treated (43°C for 10min), CT37: untreated and KD S2: shRNA SUMO-2 knock down). 1mg of lysate was used for the immunoprecipitation of SUMO-2/3 conjugates. IP experiments were performed by the protocol presented in IP and WB Method. Western blots of immunopre-cipitated proteins were developed using anti-SUMO-2/3 (Cytoskeleon cat# ASM23) (A) or anti-TFII-I antibody (B). (A) Star (*) and circle (o) indicate heavy and light chains of antibodies. Un-conjugated free SUMO is denoted by a triangle. (B) Unconjugated TFII-I is visible near 120kDa. Multiple bands indicate that TFII-I is SUMOylated by several SUMO-2/3 proteins. TFII-I has previously been reported to be a target for Sumoylation 10,11. To see the full Immunoprecipitation protocol, see the product datasheet.

Sample type

Denatured cell lysates were prepared from HS43, CT37 and KD S2

Validation comment

Supportive

Primary Ab dilution

Assay Dependent

Secondary Ab

Secondary Ab

Secondary Ab dilution

1:20000

Protocol

Protocol