720262

antibody from Invitrogen Antibodies

Targeting: MAFA hMafA, RIPE3b1

Western blot (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

720262

Provider

Invitrogen Antibodies

Product name

MAFA Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

These Polyclonal antibodies are of rabbit origin developed by immunizing animals with proteins or peptides. The polyclonal antibody is purified by affinity purification from the rabbit sera generated after immunizing the rabbits with a specific type of protein or peptide. The purified antibody is tested for its functionality in various relevant research applications. The antibody is developed for Research Use Only and is non-hazardous or non-infectious in nature.

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on nuclear extracts (30 µg lysate) of MCF-7 (Lane 1), U-2 OS (Lane 2), HEK-293 (Lane 3) and tissue extracts of Rat brain (Lane 4). The blots were probed with Anti-MAFA Rabbit Polyclonal antibody (Product # 720262, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A ~ 48 kDa band corresponding to MAFA was observed. The molecular weight of MAFA has been reported in literature to be different from its apparent molecular weight of 37 kDa. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot®2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Enrichment of endogenous MAFA protein at specific gene loci using Anti-MAFA Rabbit Polyclonal Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-MAFA Rabbit Polyclonal Antibody (Product # 720262, 3 µg) on sheared chromatin from 2 million PANC1 cells treated with 20mM of glucose for 24 hours using the MAGnify ChIP system kit (Product # 49-2024). Normal Rabbit IgG (1 µg) was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the promoters of the active INS2, SLC2A2 region used as positive control target genes, and the region of the inactive SAT2 satellite repeat, used as negative control target gene. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.