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EIF4E2 antibody [N1C3]

Anti-EIF4E2 antibody [N1C3] used in IHC (Paraffin sections) (IHC-P). GTX103977
Anti-EIF4E2 antibody [N1C3] used in Western Blot (WB). GTX103977
Anti-EIF4E2 antibody [N1C3] used in Western Blot (WB). GTX103977
Anti-EIF4E2 antibody [N1C3] used in Western Blot (WB). GTX103977

Cat No. GTX103977

Host Rabbit
Clonality Polyclonal
Isotype IgG
Application WB, IHC-P
Reactivity Human, Mouse
Package
100 μl ($319),
25 μl ($169)
APPLICATION

Application Note

*Optimal dilutions/concentrations should be determined by the researcher.
Application Dilution
WB Assay dependent
IHC-P 1:100-1:1000
Not tested in other applications.

Calculated MW

28 kDa. ( Note )

Positive Control

Target recombinant protein

Predict Reactivity

Zebrafish, Bovine, Dog, Pig, Rhesus Monkey(>80% identity)
PROPERTIES

Form

Liquid

Buffer

0.1M Tris, 0.1M Glycine, 10% Glycerol (pH7). 0.01% Thimerosal was added as a preservative.

Storage

Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.

Concentration

1 mg/ml (Please refer to the vial label for the specific concentration.)

Antigen Species

Human

Immunogen

Recombinant protein encompassing a sequence within the center region of human EIF4E2. The exact sequence is proprietary.

Purification

Purified by antigen-affinity chromatography.

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET

Synonyms

eukaryotic translation initiation factor 4E family member 2 , 4E-LP , 4EHP , EIF4EL3 , IF4e , h4EHP

Background

Recognizes and binds the 7-methylguanosine-containing mRNA cap during an early step in the initiation of protein synthesis and facilitates ribosome binding by inducing the unwinding of the mRNAs secondary structures.

Database

Research Area

DATA IMAGES
Anti-EIF4E2 antibody [N1C3] used in IHC (Paraffin sections) (IHC-P). GTX103977

GTX103977 IHC-P Image

EIF4E2 antibody [N1C3] detects EIF4E2 protein at cytoplasm on human breast carcinoma by immunohistochemical analysis.
Sample: Paraffin-embedded human breast carcinoma.
EIF4E2 antibody [N1C3] (GTX103977) diluted at 1:500.

Antigen Retrieval: Trilogy™ (EDTA based, pH 8.0) buffer, 15min

Anti-EIF4E2 antibody [N1C3] used in Western Blot (WB). GTX103977

GTX103977 WB Image

Depletion of 4EHP expression affects cell proliferation, survival, and ERK1/2 phosphorylation.(A) Cell proliferation assay. WT and 4EHP-KO MEFs were seeded in 6-well plates and trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (B) Cell proliferation assay. U251 cells with stable expression of shCTR (control), sh4EHP#1, and sh4EHP#2 were seeded in 6-well plates. Cells were trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (C) Quantitation of cell death by FACS assay; Sub-G population was considered as ‘Dead’ and G0/1, S and G2/M population was combined as ‘Live’. Data are mean ± SD (n = 3). (D) WB for the indicated proteins in the WT and 4EHP-KO MEFs. (E) Polysome profiling/RT-PCR; RNA was extracted from each fraction (collected as described in Figure 2—figure supplement 1J), subjected to electrophoresis on agarose gel and visualized, using Ethidium Bromide (EtBr) staining. RT-PCR analyses of total RNA in each fraction was carried out with primers specific for Dusp6 and Gapdh mRNAs. (F) WB on the indicated proteins in WT and 4EHP-KO MEFs. (G) WB for the indicated proteins in the WT and 4EHP-KO MEFs, expressing a v5-tagged GFP (GFP-v5) or v5-tagged 4EHP (4EHP-v5).Cell proliferation and translational regulation of DUSP6 expression is affected by 4EHP depletion.(A) WB for the indicated proteins in the WT and 4EHP-KO MEFs. (B) Cell proliferation was assessed using Sulforhodamine B (SRB ) assay . Data are mean ± SD (n = 3). (C) Top; Representative cell cycle profiles of the WT and 4EHP-KO MEFs stained with Propidium Iodide and analyzed by FACS. Bottom; quantitation of cell cycle profiles. Data are mean ± SD (n = 3). (D) WB for the indicated proteins in control and stable 4EHP-knockdown U251 cells. (E) WB for the indicated proteins in the control and stable 4EHP-knockdown U87 cells. (F) Cell proliferation assay; U87 cells with stable expression of shCTR, sh4EHP#1, and sh4EHP#2 were seeded in 6-well plates. Cells were trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (G) FACS assay. Representative cell cycle profiles of shCTR, sh4EHP#1, and sh4EHP#2 U251 cells stained with Propidium Iodide and analyzed by FACS. (H) WB for the indicated proteins in the control and stable 4EHP-knockdown U251 cells. (I) WB for the indicated proteins in the control and stable Dusp6-knockdown U251 cells. (J) Polysome profiling; cytoplasmic extract from WT and 4EHP-KO MEFs was fractioned by centrifugation on a 10–50% sucrose gradient. Fourteen fractions were collected while 254 nm absorbance was recorded. (K) WB for the indicated proteins in control (shCTR) and 4EHP-knockdown (sh4EHP) U251 cells. (L) WB for the indicated proteins in the control and stable 4EHP-knockdown U251 cells. (M) RT-qPCR analysis of Dusp6 mRNA in shCTR and sh4EHP U251 cells. Values are normalized to β-actin. Data are mean ± SD (n = 3). (N) RNA stability assay of Dusp6 mRNA in shCTR and sh4EHP U251 cells. The amount of RNA at different time points was determined by RT-qPCR. Values are normalized to 28S rRNA. Data are mean ± SD (n = 3).

Anti-EIF4E2 antibody [N1C3] used in Western Blot (WB). GTX103977

GTX103977 WB Image

Depletion of 4EHP expression affects cell proliferation, survival, and ERK1/2 phosphorylation.(A) Cell proliferation assay. WT and 4EHP-KO MEFs were seeded in 6-well plates and trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (B) Cell proliferation assay. U251 cells with stable expression of shCTR (control), sh4EHP#1, and sh4EHP#2 were seeded in 6-well plates. Cells were trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (C) Quantitation of cell death by FACS assay; Sub-G population was considered as ‘Dead’ and G0/1, S and G2/M population was combined as ‘Live’. Data are mean ± SD (n = 3). (D) WB for the indicated proteins in the WT and 4EHP-KO MEFs. (E) Polysome profiling/RT-PCR; RNA was extracted from each fraction (collected as described in Figure 2—figure supplement 1J), subjected to electrophoresis on agarose gel and visualized, using Ethidium Bromide (EtBr) staining. RT-PCR analyses of total RNA in each fraction was carried out with primers specific for Dusp6 and Gapdh mRNAs. (F) WB on the indicated proteins in WT and 4EHP-KO MEFs. (G) WB for the indicated proteins in the WT and 4EHP-KO MEFs, expressing a v5-tagged GFP (GFP-v5) or v5-tagged 4EHP (4EHP-v5).Cell proliferation and translational regulation of DUSP6 expression is affected by 4EHP depletion.(A) WB for the indicated proteins in the WT and 4EHP-KO MEFs. (B) Cell proliferation was assessed using Sulforhodamine B (SRB ) assay . Data are mean ± SD (n = 3). (C) Top; Representative cell cycle profiles of the WT and 4EHP-KO MEFs stained with Propidium Iodide and analyzed by FACS. Bottom; quantitation of cell cycle profiles. Data are mean ± SD (n = 3). (D) WB for the indicated proteins in control and stable 4EHP-knockdown U251 cells. (E) WB for the indicated proteins in the control and stable 4EHP-knockdown U87 cells. (F) Cell proliferation assay; U87 cells with stable expression of shCTR, sh4EHP#1, and sh4EHP#2 were seeded in 6-well plates. Cells were trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (G) FACS assay. Representative cell cycle profiles of shCTR, sh4EHP#1, and sh4EHP#2 U251 cells stained with Propidium Iodide and analyzed by FACS. (H) WB for the indicated proteins in the control and stable 4EHP-knockdown U251 cells. (I) WB for the indicated proteins in the control and stable Dusp6-knockdown U251 cells. (J) Polysome profiling; cytoplasmic extract from WT and 4EHP-KO MEFs was fractioned by centrifugation on a 10–50% sucrose gradient. Fourteen fractions were collected while 254 nm absorbance was recorded. (K) WB for the indicated proteins in control (shCTR) and 4EHP-knockdown (sh4EHP) U251 cells. (L) WB for the indicated proteins in the control and stable 4EHP-knockdown U251 cells. (M) RT-qPCR analysis of Dusp6 mRNA in shCTR and sh4EHP U251 cells. Values are normalized to β-actin. Data are mean ± SD (n = 3). (N) RNA stability assay of Dusp6 mRNA in shCTR and sh4EHP U251 cells. The amount of RNA at different time points was determined by RT-qPCR. Values are normalized to 28S rRNA. Data are mean ± SD (n = 3).

Anti-EIF4E2 antibody [N1C3] used in Western Blot (WB). GTX103977

GTX103977 WB Image

Depletion of 4EHP expression affects cell proliferation, survival, and ERK1/2 phosphorylation.(A) Cell proliferation assay. WT and 4EHP-KO MEFs were seeded in 6-well plates and trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (B) Cell proliferation assay. U251 cells with stable expression of shCTR (control), sh4EHP#1, and sh4EHP#2 were seeded in 6-well plates. Cells were trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (C) Quantitation of cell death by FACS assay; Sub-G population was considered as ‘Dead’ and G0/1, S and G2/M population was combined as ‘Live’. Data are mean ± SD (n = 3). (D) WB for the indicated proteins in the WT and 4EHP-KO MEFs. (E) Polysome profiling/RT-PCR; RNA was extracted from each fraction (collected as described in Figure 2—figure supplement 1J), subjected to electrophoresis on agarose gel and visualized, using Ethidium Bromide (EtBr) staining. RT-PCR analyses of total RNA in each fraction was carried out with primers specific for Dusp6 and Gapdh mRNAs. (F) WB on the indicated proteins in WT and 4EHP-KO MEFs. (G) WB for the indicated proteins in the WT and 4EHP-KO MEFs, expressing a v5-tagged GFP (GFP-v5) or v5-tagged 4EHP (4EHP-v5).Cell proliferation and translational regulation of DUSP6 expression is affected by 4EHP depletion.(A) WB for the indicated proteins in the WT and 4EHP-KO MEFs. (B) Cell proliferation was assessed using Sulforhodamine B (SRB ) assay . Data are mean ± SD (n = 3). (C) Top; Representative cell cycle profiles of the WT and 4EHP-KO MEFs stained with Propidium Iodide and analyzed by FACS. Bottom; quantitation of cell cycle profiles. Data are mean ± SD (n = 3). (D) WB for the indicated proteins in control and stable 4EHP-knockdown U251 cells. (E) WB for the indicated proteins in the control and stable 4EHP-knockdown U87 cells. (F) Cell proliferation assay; U87 cells with stable expression of shCTR, sh4EHP#1, and sh4EHP#2 were seeded in 6-well plates. Cells were trypsinized after the indicated time points and cell numbers determined using a hematocytometer. Data are mean ± SD (n = 3). (G) FACS assay. Representative cell cycle profiles of shCTR, sh4EHP#1, and sh4EHP#2 U251 cells stained with Propidium Iodide and analyzed by FACS. (H) WB for the indicated proteins in the control and stable 4EHP-knockdown U251 cells. (I) WB for the indicated proteins in the control and stable Dusp6-knockdown U251 cells. (J) Polysome profiling; cytoplasmic extract from WT and 4EHP-KO MEFs was fractioned by centrifugation on a 10–50% sucrose gradient. Fourteen fractions were collected while 254 nm absorbance was recorded. (K) WB for the indicated proteins in control (shCTR) and 4EHP-knockdown (sh4EHP) U251 cells. (L) WB for the indicated proteins in the control and stable 4EHP-knockdown U251 cells. (M) RT-qPCR analysis of Dusp6 mRNA in shCTR and sh4EHP U251 cells. Values are normalized to β-actin. Data are mean ± SD (n = 3). (N) RNA stability assay of Dusp6 mRNA in shCTR and sh4EHP U251 cells. The amount of RNA at different time points was determined by RT-qPCR. Values are normalized to 28S rRNA. Data are mean ± SD (n = 3).

REFERENCE
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Application Reference
SDS
Glycerol.pdf
Thimerosal.pdf