Recombinant Anti-HNRNPA0 antibody [EP16085] KO Tested (ab197023)

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Western blot - Anti-HNRNPA0 antibody [EP16085] (ab197023)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP16085] to HNRNPA0
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-HNRNPA0 antibody [EP16085]
See all HNRNPA0 primary antibodies
Description

Rabbit monoclonal [EP16085] to HNRNPA0
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK293T, HeLa, Jurkat, A549, 293 Mouse brain, Rat brain, C6, RAW 264.7, PC-12 and NIH/3T3 lysates. IHC-P: Human cerebral cortex and Mouse spleen tissues. ICC/IF and Flow Cyt (intra): HeLa cells.
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

EP16085
Isotype

IgG
Research areas

Alternative Versions
- Anti-HNRNPA0 antibody [EP16085] - BSA and Azide free (ab236121)
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)
KO cell lines
- Human HNRNPA0 knockout HEK-293T cell line (ab266314)
KO cell lysates
- Human HNRNPA0 knockout HEK-293T cell lysate (ab257989)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab197023 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
Flow Cyt (Intra)		1/150.
ICC/IF		1/1000.
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Detects a band of approximately 32, 34 kDa (predicted molecular weight: 31 kDa).

Notes
Flow Cyt (Intra) 1/150.
ICC/IF 1/1000.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 32, 34 kDa (predicted molecular weight: 31 kDa).

Function

This protein is a component of ribonucleosomes.
Sequence similarities

Contains 2 RRM (RNA recognition motif) domains.
Post-translational
modifications

Arg-291 is dimethylated, probably to asymmetric dimethylarginine.
Cellular localization

Nucleus. Component of ribonucleosomes.
Target information above from: UniProt accession Q13151 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10949 Human
- Entrez Gene: 77134 Mouse
- Entrez Gene: 498696 Rat
- Omim: 609409 Human
- SwissProt: Q13151 Human
- SwissProt: Q9CX86 Mouse
- Unigene: 645902 Human
- Unigene: 96996 Human
Alternative names
- Heterogeneous nuclear ribonucleoprotein A0 antibody
- hnRNA binding protein antibody
- hnRNP A0 antibody
- HNRNPA0 antibody
- HNRPA0 antibody
- ROA0_HUMAN antibody
see all

Western blot - Anti-HNRNPA0 antibody [EP16085] (ab197023)

All lanes : Anti-HNRNPA0 antibody [EP16085] (ab197023) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : HNRNPA0 knockout HEK293T cell lysate
Lane 3 : A549 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 31 kDa
Observed band size: 32,34 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab197023 observed at 32,34 kDa. Red - loading control ab8245 observed at 36 kDa.

ab197023 Anti-HNRNPA0 antibody [EP16085] was shown to specifically react with HNRNPA0 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266314 (knockout cell lysate ab257989) was used. Wild-type and HNRNPA0 knockout samples were subjected to SDS-PAGE. ab197023 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-HNRNPA0 antibody [EP16085] (ab197023)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling HNRNPA0 with ab197023 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
-ve control 1: ab197023 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNRNPA0 antibody [EP16085] (ab197023)

Immunohistochemical analysis of paraffin-embedded Human cerebral cortex labeling HNRNPA0 with ab197023 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human cerebral cortex tissue is observed. Counter stained with Hematoxylin.

Negative 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-HNRNPA0 antibody [EP16085] (ab197023)

Immunohistochemical analysis of paraffin-embedded Mouse spleen labeling HNRNPA0 with ab197023 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Mouse spleen tissue is observed. Counter stained with Hematoxylin.

Negative 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.
Flow Cytometry (Intracellular) - Anti-HNRNPA0 antibody [EP16085] (ab197023)

Intracellular Flow Cytometry analysis of HeLa cells labelling HNRNPA0 (red) with purified ab197023 at dilution of 1/150. The secondary antibody used was Alexa Fluorr^® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.
Western blot - Anti-HNRNPA0 antibody [EP16085] (ab197023)

All lanes : Anti-HNRNPA0 antibody [EP16085] (ab197023) at 1/10000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) lysate
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) lysate
Lane 3 : A549 (Human lung carcinoma) lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 31 kDa
Observed band size: 32,34 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HNRNPA0 antibody [EP16085] (ab197023)

Anti-HNRNPA0 antibody [EP16085] (ab197023) at 1/1000 dilution + 293 (Human embryonic kidney) lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 31 kDa
Observed band size: 32,34 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HNRNPA0 antibody [EP16085] (ab197023)

All lanes : Anti-HNRNPA0 antibody [EP16085] (ab197023) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : C6 (Rat glial tumor cells) lysate
Lane 4 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma) lysate
Lane 6 : NIH/3T3 (Mouse embyro fibroblast cells) lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 31 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-HNRNPA0 antibody [EP16085] (ab197023)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab197023? Please let us know so that we can cite the reference in this datasheet.

ab197023 has been referenced in 1 publication.

Zhang T et al. Silence of S1 RNA binding domain 1 represses cell growth and promotes apoptosis in human non-small cell lung cancer cells. Transl Lung Cancer Res 8:760-774 (2019). PubMed: 32010555

Customer reviews and Q&As

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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

KO cell lines

KO cell lysates

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Post-translational
modifications