Invitrogen

SLUG Polyclonal Antibody

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

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Questions & Answers

Questions & Answers

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SLUG Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of SLUG was performed using 70% confluent log phase MCF-7 cells treated with 10 ng/mL TGF-Beta for 4 days. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled Anti-SLUG Polyclonal Antibody (Product # PA5-20289) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents untreated MCF-7 cells having low expression of SLUG. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

SLUG Antibody in Western Blot (WB) — Western blot was performed using Anti-SLUG Polyclonal Antibody (Product # PA5-20289) and a 29kDa band corresponding to Slug was observed across all the cell lines tested and increased upon TGF-Beta treatment in MCF7. Whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), MDA-MB-231 (Lane 2), MCF-7 (Lane 3), MCF-7 treated with TGF-Beta (10ng/ml for 4 days (Lane 4), MCF 10A (Lane 5), U-937 (Lane 6) and Daudi were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

SLUG Antibody in Western Blot (WB) — Western Blot analysis of Slug in human kidney cell lysate with SLUG Polyclonal Antibody (Product # PA5-20289) at in (A) 1 and (B) 2 µg/mL.

SLUG Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissue tested owing to their inherent genetic constitution. Relative expression of Slug was observed in U-87 MG, MDA-MB-231, MCF-7 and MCF 10A cell lines in comparison to U-937 and Daudi cell lines which express Slug protein at lower levels using Anti-SLUG Polyclonal Antibody (Product # PA5-20289) in Western Blot {RE}

SLUG Antibody — Detection of altered expression of the target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis of Slug using Anti-SLUG Polyclonal Antibody (Product # PA5-20289), shows increased expression of Slug in MCF-7 cell line treated with TGF-Beta at a concentration of 10ng/ml for 4 days. {TM}

SLUG Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot of Slug using Anti-SLUG Polyclonal Antibody (Product # PA5-20289), shows increased expression of slug protein in MCF-7 cell line treated with TGF-Beta at a concentration of 10ng/ml for 4 days. {TM}

Product Details

PA5-20289

Applications	Tested Dilution	Publications
Western Blot (WB)	1-2 µg/mL	-
Immunocytochemistry (ICC/IF)	1:100	-

Product Specifications
Species Reactivity	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A 14 amino acid peptide from near the center of human Slug.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS
Contains	0.02% sodium azide
Storage conditions	Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C
RRID	AB_11156388

Product Specific Information

A suggested positive control is human kidney tissue lysate.

PA5-20289 can be used with blocking peptide PEP-0419.

Target Information

Slug, a member of the Snail family of C2H2-type zinc finger transcription factors, was initially identified to be involved in epithelial-mesenchymal transitions as well as the formation of the neural tube during vertebrate embryogenesis. Like Snail, Slug transcription can be induced by growth factors such as FGF, BMP, and TGF-beta. Once expressed, Slug will bind E-box regions of promoters and repress transcription of genes such as E-cadherin and Claudin-1. More recently, its expression in breast, esophogeal, and colorectal carcinomas has been correlated with poor prognosis for survival. Furthermore, Slug can protect hemopoietic progenitor cells from radiation-induced apoptosis by repressing the p53-mediated transcription of Puma, a BH3-only antagonist of the anti-apoptotic members of the Bcl-2 family.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: MGC10182; Neural crest transcription factor Slug; Protein snail homolog 2; slug (chicken homolog), zinc finger protein; slug, chicken homolog; snail family zinc finger 2; snail homolog 2; Zinc finger protein SNAI2

Gene Aliases: SLUG; SLUGH; SLUGH1; SNAI2; SNAIL2; WS2D

UniProt ID: (Human) O43623, (Mouse) P97469

Entrez Gene ID: (Human) 6591, (Mouse) 20583

Molecular Function: C2H2 zinc finger transcription factor

negative regulation of transcription from RNA polymerase II promoter osteoblast differentiation epithelial to mesenchymal transition epithelial to mesenchymal transition involved in endocardial cushion formation cell migration involved in endocardial cushion formation transcription, DNA-templated negative regulation of cell adhesion involved in substrate-bound cell migration Notch signaling pathway sensory perception of sound negative regulation of keratinocyte proliferation negative regulation of vitamin D biosynthetic process neural crest cell development positive regulation of cell migration negative regulation of chondrocyte differentiation regulation of chemokine production negative regulation of cell adhesion mediated by integrin positive regulation of histone acetylation negative regulation of catenin import into nucleus desmosome disassembly pigmentation negative regulation of DNA damage response, signal transduction by p53 class mediator cellular response to fibroblast growth factor stimulus positive regulation of fat cell differentiation regulation of osteoblast differentiation white fat cell differentiation palate development canonical Wnt signaling pathway epithelium development cartilage morphogenesis regulation of branching involved in salivary gland morphogenesis negative regulation of vitamin D receptor signaling pathway cellular response to epidermal growth factor stimulus cellular response to ionizing radiation negative regulation of canonical Wnt signaling pathway negative regulation of cell-cell adhesion by negative regulation of transcription from RNA polymerase II promoter negative regulation of intrinsic apoptotic signaling pathway in response to DNA damage negative regulation of stem cell proliferation regulation of bicellular tight junction assembly negative regulation of anoikis negative regulation of extrinsic apoptotic signaling pathway in absence of ligand regulation of transcription, DNA-templated regulation of transcription from RNA polymerase II promoter transcription from RNA polymerase II promoter multicellular organism development response to radiation cell migration cellular response to platelet-derived growth factor stimulus regulation of apoptotic process negative regulation of apoptotic process wound healing, spreading of cells positive regulation of endothelial cell chemotaxis

Process(es)

transcriptional repressor activity, RNA polymerase II core promoter proximal region sequence-specific binding chromatin binding transcription factor activity, RNA polymerase II distal enhancer sequence-specific binding protein binding sequence-specific DNA binding metal ion binding nucleic acid binding DNA binding transcription factor activity, sequence-specific DNA binding