Name: Anti-HCE antibody [EPR19384]
Brand: RabMAb
SKU: ab201046
Rating: 5 (5 reviews)

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Western blot - Anti-HCE antibody [EPR19384] (ab201046)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19384] to HCE
Suitable for: IHC-P, WB, IP, ICC/IF
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-HCE antibody [EPR19384]
See all HCE primary antibodies
Description

Rabbit monoclonal [EPR19384] to HCE
Host species

Rabbit
Tested applications

Suitable for: IHC-P, WB, IP, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal liver and fetal kidney lysates; Ramos, HepG2, SH-SY5Y, HeLa, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain and kidney lysates. IHC-P: Mouse cerebral cortex and rat spleen tissues. ICC/IF: HeLa and NIH/3T3 cells. IP: SH-SY5Y whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19384
Isotype

IgG
Research areas

Alternative Versions
- Anti-HCE antibody [EPR19384] - BSA and Azide free (ab251333)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
Recombinant Protein
- Recombinant Human HCE protein (ab183246)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab201046 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
IHC-P		1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. The IHC application is recommended for mouse and rat only.
WB	(5)	1/2000. Detects a band of approximately 69 kDa (predicted molecular weight: 69 kDa).
IP		1/50.
ICC/IF		1/500.

Notes
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. The IHC application is recommended for mouse and rat only.
WB 1/2000. Detects a band of approximately 69 kDa (predicted molecular weight: 69 kDa).
IP 1/50.
ICC/IF 1/500.

Function

Bifunctional mRNA-capping enzyme exhibiting RNA 5'-triphosphatase activity in the N-terminal part and mRNA guanylyltransferase activity in the C-terminal part. Catalyzes the first two steps of cap formation: by removing the gamma-phosphate from the 5'-triphosphate end of nascent mRNA to yield a diphosphate end, and by transferring the gmp moiety of GTP to the 5'-diphosphate terminus.
Tissue specificity

Isoform 1 and isoform 4 (at a lesser extent) are expressed in cerebrum, cerebellum, thyroid, lung, heart, liver, kidney, spleen, large intestine, testis, skin and muscle.
Sequence similarities

In the N-terminal section; belongs to the non-receptor class of the protein-tyrosine phosphatase family.
In the C-terminal section; belongs to the eukaryotic GTase family.
Cellular localization

Nucleus.
Target information above from: UniProt accession O60942 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 8732 Human
- Entrez Gene: 24018 Mouse
- Entrez Gene: 313131 Rat
- Omim: 603512 Human
- SwissProt: O60942 Human
- SwissProt: O55236 Mouse
- Unigene: 567378 Human
- Unigene: 240024 Mouse
- Unigene: 464346 Mouse
see all
Alternative names
- CAP1A antibody
- GTase antibody
- GTP--RNA guanylyltransferase antibody
- hCAP antibody
- HCAP1 antibody
- HcE antibody
- HCE1 antibody
- MCE1 antibody
- MCE1_HUMAN antibody
- mRNA 5''-triphosphatase antibody
- mRNA capping enzyme antibody
- mRNA guanyltransferase antibody
- mRNA guanylyltransferase antibody
- Polynucleotide 5' trisphosphatase antibody
- RNA guanylyltransferase and 5' phosphatase antibody
- RNGTT antibody
- TPase antibody
see all

Western blot - Anti-HCE antibody [EPR19384] (ab201046)

All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 69 kDa
Observed band size: 69 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HCE antibody [EPR19384] (ab201046)

All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

Lane 1 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 3 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 69 kDa
Observed band size: 69 kDa

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HCE antibody [EPR19384] (ab201046)

All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 69 kDa
Observed band size: 69 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HCE antibody [EPR19384] (ab201046)

All lanes : Anti-HCE antibody [EPR19384] (ab201046) at 1/2000 dilution

Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 69 kDa
Observed band size: 69 kDa

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCE antibody [EPR19384] (ab201046)

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling HCE with ab201046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on neurons of mouse cerebral cortex is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HCE antibody [EPR19384] (ab201046)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling HCE with ab201046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on lymphocytes of rat spleen is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-HCE antibody [EPR19384] (ab201046)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HCE with ab201046 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [EPR19384] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab201046 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-HCE antibody [EPR19384] (ab201046)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HCE with ab201046 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [EPR19384] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab201046 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Immunoprecipitation - Anti-HCE antibody [EPR19384] (ab201046)

HCE was immunoprecipitated from 1mg of SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate with ab201046 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab201046 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: SH-SY5Y whole cell lysate 10µg (Input).
Lane 2: ab201046 IP in SH-SY5Y whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR19384] -Isotype Control (ab172730) instead of ab201046 in SH-SY5Y whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
Anti-HCE antibody [EPR19384] (ab201046)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab201046? Please let us know so that we can cite the reference in this datasheet.

ab201046 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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1-5 of 5 Abreviews

Application

Western blot

Sample

Dog Cell lysate - whole cell (MDCK cells)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

MDCK cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 05 2022

Application

Western blot

Sample

Monkey Cell lysate - whole cell (COS-1 cells)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

COS-1 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 03 2022

Application

Western blot

Sample

Rat Cell lysate - whole cell (3611-RF cells)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

3611-RF cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 03 2022

Application

Western blot

Sample

Mouse Cell lysate - whole cell (MEFs)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

MEFs

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 03 2022

Application

Western blot

Sample

Human Cell lysate - whole cell (HeLa cells)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

HeLa cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 03 2022

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

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