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29650
Ferroptosis Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Ferroptosis Antibody Sampler Kit #29650

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Ferroptosis Antibody Sampler Kit: Image 1
Western blot analysis of extracts from Huh7 and Hep G2 cells using xCT/SLC7A11 (D2M7A) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 2
Western blot analysis of extracts from MEF wt and U-2 OS cells, untreated (-) or treated with MG-132 #2194 (10 μM, 10 hr; +), using NRF2 (D1Z9C) XP®Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 3
Western blot analysis of extracts from SK-MEL-5 and SK-MEL-2 cells using DMT1/SLC11A2 (D3V8G) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 4
Western blot analysis of extracts from HeLa and HT-29 cells using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 5
Western blot analysis of extracts from various cell lines using 4F2hc/CD98 (D3F9D) XP® Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower).
Ferroptosis Antibody Sampler Kit: Image 6
Western blot analysis of extracts from various cell lines using GPX4 Antibody.
Ferroptosis Antibody Sampler Kit: Image 7
Western blot analysis of extracts from various cell lines using NCOA4 (E8H8Z) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, HT-1080 and OVCAR-4 cells are low for NCOA4 expression.
Ferroptosis Antibody Sampler Kit: Image 8
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.
Ferroptosis Antibody Sampler Kit: Image 9
Western blot analysis of extracts from various cell lines using KEAP1 (D6B12) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 10
Immunoprecipitation of NRF2 from MEF wt cell extracts treated with MG-132 #2194 (10 μM, 10 hr) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or NRF2 (D1Z9C) XP®Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using NRF2 (D1Z9C) XP®Rabbit mAb (lane 3).
Ferroptosis Antibody Sampler Kit: Image 11
Western blot analysis of extracts from human SK-MEL-5 cells, untreated (-) or treated with PNGase F (+), using DMT1/SLC11A2 (D3V8G) Rabbit mAb (upper) and β-Actin (13E5) Rabbit mAb #4970 (lower).
Ferroptosis Antibody Sampler Kit: Image 12
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using 4F2hc/CD98 (D3F9D) XP®Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 14
Western blot analysis of extracts from mouse testis using GPX4 Antibody.
Ferroptosis Antibody Sampler Kit: Image 15
Immunoprecipitation of NCOA4 protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NCOA4 (E8H8Z) Rabbit mAb. Western blot analysis was performed using NCOA4 (E8H8Z) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Ferroptosis Antibody Sampler Kit: Image 16
Western blot analysis of extracts from OVCAR8 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence®KEAP1 siRNA I #5285 (+) or SignalSilence® KEAP1 siRNA II #5289 (+), using KEAP1 (D6B12) Rabbit mAb (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The KEAP1 (D6B12) Rabbit mAb confirms silencing of KEAP1 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.
Ferroptosis Antibody Sampler Kit: Image 17
Confocal immunofluorescent analysis of wild-type NRF2 MEF cells, untreated (left) or treated with MG-132 #2194 (10 μM, 8 hr; center), and NRF2 knock-out MEF cells treated with MG-132 #2194 (10 μM, 8 hr; right), using NRF2 (D1Z9C) XP® Rabbit mAb (green pseudocolor). Actin filaments were labeled with Alexa Fluor® 488 Phalloidin #8878 (red pseudocolor).
Ferroptosis Antibody Sampler Kit: Image 18
Immunoprecipitation of DMT1 from SK-MEL-5 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or DMT1/SLC11A2 (D3V8G) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using DMT1/SLC11A2 (D3V8G) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded HeLa cell pellet (left, high-expressing) or HL-60 cell pellet (right, low-expressing) using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using 4F2hc/CD98 (D3F9D) XP®Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 21
Confocal immunofluorescent analysis of OVCAR8 cells using KEAP1 (D6B12) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Ferroptosis Antibody Sampler Kit: Image 22
Flow cytometric analysis of MEF wt cells, untreated (blue) or treated with MG-132 #2194 (10uM, 4 hrs; green) using NRF2 (D1Z9C) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Ferroptosis Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using FTH1 (D1D4) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Ferroptosis Antibody Sampler Kit: Image 24
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using 4F2hc/CD98 (D3F9D) XP®Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 25
Immunohistochemical analysis of paraffin-embedded normal human lung using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 26
Chromatin immunoprecipitations were performed with cross-linked chromatin from MEF cells treated with DEM (50 μM, 3 hr) and NRF2 (D1Z9C) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across NQO1, a known target gene of NRF2 (see additional figure containing ChIP-qPCR data).
Ferroptosis Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 28
Confocal immunofluorescent analysis of HeLa (left, high expressing) or SH-SY5Y (right, low expressing) cells using 4F2hc/CD98 (D3F9D) XP®Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Ferroptosis Antibody Sampler Kit: Image 29
Chromatin immunoprecipitations were performed with cross-linked chromatin from MEF cells treated with DEM (50 μM, 3 hr) and NRF2 (D1Z9C) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 8 (upper), including NQO1 (lower), a known target gene of NRF2 (see additional figure containing ChIP-qPCR data).
Ferroptosis Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded normal mouse colon using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 31
Flow cytometric analysis of SH-SY5Y cells (blue) and HeLa cells (green) using 4F2hc/CD98 (D3F9D) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Ferroptosis Antibody Sampler Kit: Image 32
Chromatin immunoprecipitations were performed with cross-linked chromatin from MEF NRF2 wild-type (left) and NRF2 knock-out (right) cells, both treated with DEM (50 μM, 3 hr), and NRF2 (D1Z9C) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using mouse MafG intron 1 primers, SimpleChIP® Mouse NQO1 Promoter Primers #12635, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Ferroptosis Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded normal mouse liver using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 34
Immunohistochemical analysis of paraffin-embedded normal mouse spleen using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 35
Immunohistochemical analysis of paraffin-embedded normal mouse testis using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 36
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 37
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using FTH1 (D1D4) Rabbit mAb.
Ferroptosis Antibody Sampler Kit: Image 38
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using FTH1 (D1D4) Rabbit mAb.
To Purchase # 29650T
Product # Size Price
29650T
1 Kit  (8 x 20 microliters) $ 570

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
GPX4 Antibody 52455 20 µl
  • WB
H M Mk 20, 22 Rabbit 
NCOA4 (E8H8Z) Rabbit mAb 66849 20 µl
  • WB
  • IP
H 80 Rabbit IgG
KEAP1 (D6B12) Rabbit mAb 8047 20 µl
  • WB
  • IF
H M R 60-64 Rabbit IgG
NRF2 (D1Z9C) XP®Rabbit mAb 12721 20 µl
  • WB
  • IP
  • IF
  • F
  • ChIP
H M Mk 97-100 Rabbit IgG
4F2hc/CD98 (D3F9D) XP® Rabbit mAb 47213 20 µl
  • WB
  • IHC
  • IF
  • F
H 75-120 Rabbit IgG
FTH1 (D1D4) Rabbit mAb 4393 20 µl
  • WB
  • IP
  • IHC
H M R Mk 21 Rabbit IgG
xCT/SLC7A11 (D2M7A) Rabbit mAb 12691 20 µl
  • WB
  • IP
H 35 Rabbit IgG
DMT1/SLC11A2 (D3V8G) Rabbit mAb 15083 20 µl
  • WB
  • IP
H 55, 70-100 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Ferroptosis Antibody Sampler Kit provides an economical means of detecting proteins involved in ferroptosis. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Ferroptosis Antibody Sampler Kit detects endogenous levels of its target protein. 4F2hc/CD98 (D3F9D) XP® Rabbit mAb is predicted to detect multiple isoforms of 4F2hc/CD98.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Gly213 of human NCOA4, Ala275 of human NRF2, Val509 of human 4F2hc/CD98, Ala224 of human xCT/SLC7A11, residues near the carboxy terminus of human KEAP1 and human FTH1, and residues near the amino terminus of human DMT1. GPX4 Antibody is a polyclonal antibody produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human GPX4 protein. Antibodies are purified by peptide affinity chromatography.

Background

Ferroptosis is an iron-dependent form of regulated cell death associated with an increase in lipid peroxides (reviewed in 1,2). Free divalent iron (Fe2+) can lead to spontaneous lipid peroxidation through a Fenton reaction. Ferroptosis is regulated by signaling pathways that control iron storage and oxidative stress. Iron homeostasis is controlled, in part, by ferritin, an iron storage protein consisting of a complex of heavy (FTH1) and light (FTL) chains. Levels of ferritin may be regulated by a selective autophagy process targeting ferritin, termed ferritinophagy. This pathway is mediated by nuclear receptor coactivator 4 (NCOA4), a selective cargo receptor for ferritin (3,4). The divalent metal transporter SLC11A2/DMT1/NRAMP2 regulates iron homeostasis through non-heme absorption in the intestine (5). The glutathione peroxidase pathway has been identified as a key antioxidant defense pathway triggering ferroptosis. The compound RSL3, which directly inhibits GPX4, was identified as an activator of ferroptosis (6). GPX4 converts GSH into oxidized glutathione (GSSH) and reduces cytotoxic lipid peroxides. The glutathione peroxidase pathway is further regulated by System Xc-, an amino acid antiporter consisting of a disulfide-linked heterodimer of SLC7A11/xCT and SLC3A2/4F2hc/CD98, and is inhibited by the ferroptosis inducer erastin (7). Regulation of genes involved in oxidative stress, including GPX4, are largely controlled by the transcription factor NRF2 and serves as a defense against ferroptosis (8). Under normal conditions, expression of NRF2 is inhibited through interaction with KEAP1, part of a ubiquitin E3 ligase complex that leads to NRF2 proteasomal degradation. Oxidative stress leads to conformational changes in KEAP1 that disrupts this interaction, resulting in stabilization of NRF2. This process is further regulated through the autophagy pathway in which the autophagy cargo receptor p62/SQSTM1 can competitively inhibit the KEAP1-NRF2 complex, leading to upregulation of NRF2.
  1. Cao, J.Y. and Dixon, S.J. (2016) Cell Mol Life Sci 73, 2195-209.
  2. Xie, Y. et al. (2016) Cell Death Differ 23, 369-79.
  3. Mancias, J.D. et al. (2014) Nature 509, 105-9.
  4. Dowdle, W.E. et al. (2014) Nat Cell Biol 16, 1069-79.
  5. Gunshin, H. et al. (1997) Nature 388, 482-8.
  6. Yang, W.S. et al. (2014) Cell 156, 317-31.
  7. Dixon, S.J. et al. (2014) Elife 3, e02523.
  8. Fan, Z. et al. (2017) Oncogenesis 6, e371.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
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