Revision 4

#38493

Store at -20C

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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 M R

Sensitivity:
Endogenous

MW (kDa):
80

Source/Isotype:
Rabbit IgG

UniProt ID:
#O60488

Entrez-Gene Id:
2182

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:200

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

ACSL4 (F6T3Z) Rabbit Monoclonal Antibody recognizes endogenous levels of total ACSL4 protein. Species reactivity for immunofluorescence is human only.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human ACSL4 protein.

Background

The long-chain fatty acyl-CoA synthetase (ACSL) family of enzymes convert free long-chain fatty acids into their active form, acyl-coenzyme A (acyl-CoA), and play dominant roles in both anabolic (fatty acid synthesis and lipogenesis) and catabolic (lipolysis and fatty acid β-oxidation) pathways (1). Acyl-CoA synthetase long-chain family member 4 (ACSL4) preferentially catalyzes the formation of arachidonoyl-coenzyme A (AA-CoA) by inserting coenzyme A (CoA) into arachidonic acid (AA) (1). ACSL4-catalyzed acyl-CoAs also participate in the regulation of steroidogenesis (2), eicosanoid biosynthesis (3), and phospholipid remodeling (4). ACSL4 is an essential enzyme for the conversion of two key ferroptosis-inducing signals, oxidized arachidonic and adrenic phosphatidylethanolamines (5,6). Genome-wide CRISPR/Cas9-based genetic screens have demonstrated that ACSL4 is an essential component for ferroptosis execution (6), and genetic or pharmacological inhibition of ACSL4 can initiate a specific anti-ferroptotic rescue pathway (5). ACSL4 expression is upregulated in ferroptosis-sensitive cancer cells compared with ferroptosis-resistant cells (7) and has been shown to predict sensitivity to ferroptosis in a panel of basal-like breast cancer cell lines (6). Moreover, high ACSL4 protein expression in hepatocellular carcinoma (HCC) clinical tissue specimens is associated with improved patient outcomes after sorafenib treatment and could serve as a predictive biomarker (8).

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 nonfat dry milk, 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 M: Mouse R: Rat

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.

使用に関する制限

使用に関する制限

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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 4

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using ACSL4 (F6T3Z) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of ACSL4 protein in MCF7 and T-47D cells is consistent with the predicted expression pattern.
Western Blotting Image 1: ACSL4 (F6T3Z) Rabbit Monoclonal Antibody
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human ACSL4 protein (hACSL4; +), using ACSL4 (F6T3Z) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: ACSL4 (F6T3Z) Rabbit Monoclonal Antibody
Simple Western™️ analysis of lysates (0.05mg/mL) from Huh-7 cells using ACSL4 (F6T3Z) Rabbit mAb #38493. 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 Jess™️ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12 – 230 kDa separation module.
Western Blotting Image 1: ACSL4 (F6T3Z) 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 4

Cell Signaling Technology Logo
Immunoprecipitation of ACSL4 protein from HuH-7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is ACSL4 (F6T3Z) Rabbit mAb. Western blot analysis was performed using ACSL4 (F6T3Z) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Immunoprecipitation Image 1: ACSL4 (F6T3Z) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of Caki-1 cells (left, positive) and MCF7 cells (right, negative) using ACSL4 (F6T3Z) Rabbit mAb (green), DyLight 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
Immunofluorescence Image 1: ACSL4 (F6T3Z) 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.