Revision 1

#26118

Store at -20C

Hedgehog Signaling Antibody Sampler Kit

1 Kit

(8 x 20 microliters)

CST Logo
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.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Shh (C9C5) Rabbit Monoclonal Antibody220720 µl19, (45 precursor) kDaRabbit IgG
Smoothened (E6Z5T) Rabbit Monoclonal Antibody9298120 µl85 kDaRabbit IgG
PTCH1 (C53A3) Rabbit Monoclonal Antibody246820 µl180-210 kDaRabbit IgG lambda
PTCH2 (G1191) Antibody247020 µl130 kDaRabbit
SUFU (C54G2) Rabbit Monoclonal Antibody252020 µl54 kDaRabbit IgG
GLI1 (C68H3) Rabbit Monoclonal Antibody353820 µl160 kDaRabbit IgG
GLI2 (E5V8N) Rabbit Monoclonal Antibody1877320 µl220 kDaRabbit IgG
GLI3 (E6E2K) Rabbit Monoclonal Antibody7110720 µl200 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Hedgehog Signaling Antibody Sampler Kit provides an economical means of evaluating key members of the Hedgehog signaling pathway. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

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 antibodies.

Background

The Hedgehog (Hh) signaling pathway plays critical roles in the regulation of cell fate, tissue patterning, and growth during embryonic development. It is downregulated during postnatal development, but can be reactivated to promote tissue repair and regeneration. Aberrant Hh signaling activity during prenatal development is associated with numerous birth defects (e.g., holoprosencephaly), while uncontrolled Hh pathway activation postnatally is linked to the development of several cancer types (1,2). There are three canonical Hh ligands: Sonic hedgehog (SHH), Indian hedgehog (IHH), and Desert hedgehog (DHH), all of which have distinct as well as overlapping roles and expression patterns (3-5). Patched1 and 2 (PTCH1 and PTCH2) are partially redundant 12-pass transmembrane proteins that function as receptors for Hh ligands (6-8). Smoothened (SMO) is a 7-pass transmembrane G protein-coupled receptor (GPCR) that functions as the key transducer of Hh signaling. In the absence of Hh ligands (off-state), PTCH proteins are localized to cilia, and function to suppress SMO activity (1,2). Suppressor of Fused (SUFU) simultaneously contributes to suppression of the pathway by sequestering the glioma-associated oncogene (GLI) family of transcription factors (9,10). Binding of Hh ligands to PTCH receptors results in derepression of SMO, in part by promoting its translocation to cilia; this leads to downregulation of SUFU, resulting in the stabilization and nuclear translocation of GLI transcription factors that regulate the transcription of genes involved in cell proliferation, migration, and survival (1).

Trademarks and Patents

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

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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 1

Cell Signaling Technology Logo
Immunoprecipitation of GLI2 protein from SNU-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is GLI2 (E5V8N) Rabbit mAb. Western blot analysis was performed using GLI2 (E7R1N) Rabbit mAb #66670. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
undefined Image 1: Hedgehog Signaling Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using GLI2 (E5V8N) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). As expected, BEN cells are negative for GLI2 expression.
undefined Image 2: Hedgehog Signaling Antibody Sampler Kit
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged GLI2 protein (lane 2), using GLI2 (E5V8N) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 3: Hedgehog Signaling Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from HT29 and GH3 cells, and GH3 cell conditioned medium (CM), using Shh (C9C5) Rabbit mAb.
undefined Image 4: Hedgehog Signaling Antibody Sampler Kit
Western blot analysis of total cell lysates from COS cells, untransfected or transiently transfected with a human PTCH1 expression construct, using PTCH1 (C53A3) Rabbit mAb.
undefined Image 5: Hedgehog Signaling Antibody Sampler Kit
Western blot analysis of extracts from COS cells, untransfected or transiently transfected with a construct expressing human PTCH2, using PTCH2 (G1191) Antibody.
undefined Image 6: Hedgehog Signaling Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Western blot analysis of total cell lysates from various cell types using SUFU (C54G2) Rabbit mAb.
undefined Image 7: Hedgehog Signaling Antibody Sampler Kit
Western blot analysis of extracts from RMS-13 and TOV-112D cells using GLI1 (C68H3) Rabbit mAb.
undefined Image 8: Hedgehog Signaling Antibody Sampler Kit
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 9: Hedgehog Signaling Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using GLI3 (E6E2K) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of GLI3 protein in NCI-H146 and Hep G2 cells is consistent with predicted expression patterns, confirming specificity of the antibody for GLI3.
undefined Image 10: Hedgehog Signaling Antibody Sampler Kit
Immunoprecipitation of Smoothened protein from extracts of 293T cells transiently transfected with plasmid encoding Myc/DDK-tagged Smoothened protein. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Smoothened (E6Z5T) Rabbit mAb. Western blot analysis was performed using Smoothened (E6Z5T) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
undefined Image 11: Hedgehog Signaling Antibody Sampler Kit
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged Smoothened protein (lane 2), using Smoothened (E6Z5T) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). *The antibody detects 25 kDa and 38 kDa proteins of unknown identity in some cell extracts.
undefined Image 12: Hedgehog Signaling Antibody Sampler Kit
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 1

Cell Signaling Technology Logo
Immunoprecipitation of endogenous PTCH1 from 293 cell lysates using PTCH1 (C53A3) Rabbit mAb. Western blot detection was performed using the same antibody.
undefined Image 13: Hedgehog Signaling Antibody Sampler Kit
Immunoprecipitation of GLI3 protein from RD cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is GLI3 (E6E2K) Rabbit mAb. Western blot analysis was performed using GLI3 (E6E2K) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
undefined Image 14: Hedgehog Signaling Antibody Sampler Kit
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.