Name: Anti-HNRPAB antibody [EPR16944]
Brand: RabMAb
SKU: ab199724
Rating: 4 (1 reviews)

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Western blot - Anti-HNRPAB antibody [EPR16944] (ab199724)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16944] to HNRPAB
Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IF
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 647 Carrier Free

Product name

Anti-HNRPAB antibody [EPR16944]
See all HNRPAB primary antibodies
Description

Rabbit monoclonal [EPR16944] to HNRPAB
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), IP, IHC-P, WB, ICC/IFmore 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, K562, Jurkat, HeLa, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal brain lysate; Mouse brain and rat brain lysates. IHC-P: Human cervix carcinoma, mouse liver and rat cardiac muscle tissues. ICC/IF: Jurkat cells. IP: K562 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

EPR16944
Isotype

IgG
Research areas

Alternative Versions
- Alexa Fluor® 647 Anti-HNRPAB antibody [EPR16944] (ab210053)
- Anti-HNRPAB antibody [EPR16944] - BSA and Azide free (ab251289)
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 HNRNPAB knockout HEK-293T cell line (ab266459)
KO cell lysates
- Human HNRNPAB knockout HEK-293T cell lysate (ab257467)
Recombinant Protein
- Recombinant Human HNRPAB protein (ab171508)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab199724 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)		Use a concentration of 1 µg/ml.
IP		1/30.
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB	(1)	1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 36 kDa).
ICC/IF		1/500.

Notes
Flow Cyt (Intra) Use a concentration of 1 µg/ml.
IP 1/30.
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 42 kDa (predicted molecular weight: 36 kDa).
ICC/IF 1/500.

Function

Binds single-stranded RNA. Has a high affinity for G-rich and U-rich regions of hnRNA. Also binds to APOB mRNA transcripts around the RNA editing site.
Tissue specificity

Ubiquitous.
Sequence similarities

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

Dimethylation at Arg-322 is probably asymmetric.
Cellular localization

Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
Target information above from: UniProt accession Q99729 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3182 Human
- Entrez Gene: 15384 Mouse
- Entrez Gene: 83498 Rat
- Omim: 602688 Human
- SwissProt: Q99729 Human
- SwissProt: Q99020 Mouse
- Unigene: 591731 Human
- Unigene: 715055 Human
- Unigene: 256875 Mouse
- Unigene: 280842 Mouse
see all
Alternative names
- ABBP 1 antibody
- ABBP-1 antibody
- ABBP1 antibody
- Apobec 1 binding protein 1 antibody
- APOBEC1 binding protein 1 antibody
- APOBEC1-binding protein 1 antibody
- Apolipoprotein B mRNA editing enzyme catalytic polypeptide 1- protein 1 antibody
- FLJ40338 antibody
- Heterogeneous nuclear ribonucleoprotein A/B antibody
- hnRNP A/B antibody
- hnRNP type A/B protein antibody
- HNRNPAB antibody
- HNRPAB antibody
- ROAA_HUMAN antibody
see all

Western blot - Anti-HNRPAB antibody [EPR16944] (ab199724)

All lanes : Anti-HNRPAB antibody [EPR16944] (ab199724) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : HNRNPAB knockout HEK293T cell lysate
Lane 3 : K-562 cell lysate
Lane 4 : Jurkat 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: 36 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab199724 observed at 48 kDa. Red - loading control ab8245 observed at 36 kDa.

ab199724 Anti-HNRPAB antibody [EPR16944] was shown to specifically react with HNRPAB in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266459 (knockout cell lysate ab257467) was used. Wild-type and HNRPAB knockout samples were subjected to SDS-PAGE. ab199724 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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.
Western blot - Anti-HNRPAB antibody [EPR16944] (ab199724)

All lanes : Anti-HNRPAB antibody [EPR16944] (ab199724) at 1/5000 dilution

Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell 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: 36 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

The expression profile observed is consistent with what has been described in the literature PMID: 22332140.
Blocking/Dilution buffer: 5% NFDM/TBST.
Flow Cytometry (Intracellular) - Anti-HNRPAB antibody [EPR16944] (ab199724)

ab199724 staining HNRPAB in Jurkat (human acute T cell leukemia) cellsby intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/2700. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Western blot - Anti-HNRPAB antibody [EPR16944] (ab199724)

Anti-HNRPAB antibody [EPR16944] (ab199724) at 1/5000 dilution + Human fetal brain lysate at 20 µg

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

Predicted band size: 36 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

The expression profile observed is consistent with what has been described in the literature PMID: 22332140.
Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HNRPAB antibody [EPR16944] (ab199724)

All lanes : Anti-HNRPAB antibody [EPR16944] (ab199724) at 1/1000 dilution

Lane 1 : Mouse brain lysates
Lane 2 : Rat brain lysates
Lane 3 : C6 (Rat glial tumor cells) whole cell lysate
Lane 4 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 6 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell 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: 36 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

The expression profile observed is consistent with what has been described in the literature PMID: 22332140.
Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNRPAB antibody [EPR16944] (ab199724)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling HNRPAB with ab199724 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human cervix carcinoma tissue 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).

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-HNRPAB antibody [EPR16944] (ab199724)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling HNRPAB with ab199724 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on mouse liver tissue 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).

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-HNRPAB antibody [EPR16944] (ab199724)

Immunohistochemical analysis of paraffin-embedded rat cardiac muscle tissue labeling HNRPAB with ab199724 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on rat cardiac muscle tissue 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).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-HNRPAB antibody [EPR16944] (ab199724)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling HNRPAB with ab199724 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on Jurkat cell line is observed. The nuclear counterstain 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: ab199724 at 1/500 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 1/500 dilution.
Immunoprecipitation - Anti-HNRPAB antibody [EPR16944] (ab199724)

HNRPAB was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab199724 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199724 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: K562 whole cell lysate 10 µg (Input). Lane 2: ab199724 IP in K562 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199724 in K562 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Anti-HNRPAB antibody [EPR16944] (ab199724)

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

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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Publishing research using ab199724? Please let us know so that we can cite the reference in this datasheet.

ab199724 has been referenced in 1 publication.

Zhou JM et al. High hnRNP AB expression is associated with poor prognosis in patients with colorectal cancer. Oncol Lett 18:6459-6468 (2019). PubMed: 31819776

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Application

Western blot

Sample

Xenopus laevis Cell lysate - other (oocytes)

Gel Running Conditions

Reduced Denaturing (8% TrisGlycine Gel)

Loading amount

0.1 µg

Specification

oocytes

Blocking step

Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°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.

DR. Samantha Jeschonek

Verified customer

Submitted Jan 09 2018

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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

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

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

Western blot abreview for Anti-HNRPAB antibody [EPR16944]

Post-translational
modifications