Revision 3

#80075

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP, IF-IC

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
80, 90, 130

Source/Isotype:
Rabbit IgG

UniProt ID:
#P23497

Entrez-Gene Id:
6672

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:6400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

SP100 (F1H6X) Rabbit Monoclonal Antibody recognizes endogenous levels of total SP100 protein. This antibody recognizes 7 isoforms of SP100, including SP100-HMG, SP100-A, SP100-B, and SP100-C. This antibody detects a 20 kDa protein of unknown identity in some cell lines.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro382 of human SP100 protein.

Background

Speckled protein 100 kDa (SP100), also known as SP100 nuclear antigen, was first identified as an antigen reactive with antibodies from patients with autoimmune disorders (1,2). The nuclear distribution of SP100 predominantly colocalizes with promyelocytic leukemia-nuclear bodies (PML-NBs) (3), which function as structures that regulate several diverse cellular functions, including gene regulation, DNA-damage responses, and apoptosis (4-6). This protein contains several isoforms, and the major isoform SP100-A has a calculated molecular weight of 54 kDa but exerts aberrant electrophoretic mobilities in SDS-polyacrylamide gel electrophoresis of approximately 100 kDa (1,2). SP100 has also been shown to undergo extensive post-translational modifications (PTM), including phosphorylation, ubiquitination, and SUMOylation. Moreover, PML-NBs play an important role in the PTM status of SP100, since they are thought to serve as hotspots for SUMOylation reactions, and PML depletion reduces SP100 SUMOylation (7-9).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using SP100 (F1H6X) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Negative expression of SP100 protein in 293T cells is consistent with the predicted expression pattern.
Western Blotting Image 1: SP100 (F1H6X) Rabbit Monoclonal Antibody
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc-tagged full-length human SP100-HMG (hSP100-Myc; +) using SP100 (F1H6X) Rabbit mAb, Myc-Tag (71D10) Rabbit mAb #2278 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: SP100 (F1H6X) Rabbit Monoclonal Antibody
Simple WesternTM analysis of lysates (0.1 mg/mL) from Raji using SP100 (F1H6X) Rabbit mAb #80075. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12 - 230 kDa separation module.
Western Blotting Image 1: SP100 (F1H6X) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Immunoprecipitation of SP100 protein from SK-MEL-30 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SP100 (F1H6X) Rabbit mAb. Western blot analysis was performed using SP100 (F1H6X) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: SP100 (F1H6X) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of SK-MEL-30 cells (left, positive) and TOV-112D cells (right, negative) using SP100 (F1H6X) Rabbit mAb (green) and DAPI #4083 (blue).
Immunofluorescence Image 1: SP100 (F1H6X) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.