Name: Anti-HRAS + KRAS antibody [EPR18713]
Brand: RabMAb
SKU: ab191595
Rating: 3 (5 reviews)

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Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18713] to HRAS + KRAS
Suitable for: ICC/IF, WB, IP
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-HRAS + KRAS antibody [EPR18713]
See all HRAS + KRAS primary antibodies
Description

Rabbit monoclonal [EPR18713] to HRAS + KRAS
Host species

Rabbit
Specificity

IP: Immunoprecipitation requires a mild lysis buffer to achieve an acceptable enrichment.
Tested applications

Suitable for: ICC/IF, WB, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human KRAS and HRAS full length proteins; Human fetal kidney and fetal brain lysates; Mouse and Rat brain lysates; A549, C6, A431 and RAW 264.7 whole cell lysates. ICC/iF: HeLa and NIH/3T3 cells. IP: A549 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

EPR18713
Isotype

IgG
Research areas

Alternative Versions
- Anti-HRAS + KRAS antibody [EPR18713] - BSA and Azide free (ab240323)
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)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab191595 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
ICC/IF		1/500.
WB	(2)	1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 22, 21 kDa).
IP	(2)	1/70. Immunoprecipitation requires a mild lysis buffer to achieve an acceptable enrichment.

Notes
ICC/IF 1/500.
WB 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 22, 21 kDa).
IP 1/70. Immunoprecipitation requires a mild lysis buffer to achieve an acceptable enrichment.

Relevance

KRAS: Ras proteins bind GDP/GTP and possess intrinsic GTPase activity. Plays an important role in the regulation of cell proliferation (PubMed:23698361, PubMed:22711838). Enzyme regulation: Alternates between an inactive form bound to GDP and an active form bound to GTP. Activated by a guanine nucleotide-exchange factor (GEF) and inactivated by a GTPase-activating protein (GAP). Interaction with SOS1 promotes exchange of bound GDP by GTP. HRAS: Ras proteins bind GDP/GTP and possess intrinsic GTPase activity. Enzyme regulation: Alternates between an inactive form bound to GDP and an active form bound to GTP. Activated by a guanine nucleotide-exchange factor (GEF) and inactivated by a GTPase-activating protein (GAP).
Database links
- Entrez Gene: 3265 Human
- Entrez Gene: 3845 Human
- Omim: 190020 Human
- Omim: 190070 Human
- SwissProt: P01112 Human
- SwissProt: P01116 Human
- SwissProt: P32883 Mouse
- SwissProt: Q61411 Mouse
- SwissProt: P08644 Rat
- SwissProt: P20171 Rat
see all
Alternative names
- c-H-ras antibody
- c-K-ras antibody
- c-Ki-ras antibody
- GTPase HRas antibody
- GTPase KRas antibody
- H-Ras-1 antibody
- Ha-Ras antibody
- HRAS antibody
- HRAS1 antibody
- K-Ras 2 antibody
- Ki-Ras antibody
- KRAS antibody
- KRAS2 antibody
- p21ras antibody
- RASK2 antibody
- Transforming protein p21 antibody
see all

Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

All lanes : Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/10000 dilution

Lane 1 : Human KRAS full length protein
Lane 2 : Human HRAS full length protein
Lane 3 : Human NRAS full length protein

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.
Human KRAS full length protein contains aa1-189 with His-Tag® at N-Terminus; Human HRAS full length protein contains aa1-189 with His-Tag® at N-Terminus; Human NRAS full length protein contains aa1-189 with His-Tag® at N-Terminus. These are all made in-house.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/2000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/5000 dilution + Human fetal brain lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

All lanes : Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/2000 dilution + A549 (Human lung carcinoma) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

All lanes : Anti-HRAS + KRAS antibody [EPR18713] (ab191595) at 1/5000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysate
Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 22, 21 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling KRAS + HRAS with ab191595 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasm and nuclear staining on HeLa cell line.
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 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab191595 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 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 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling KRAS + HRAS with ab191595 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasm and nuclear staining on NIH/3T3 cells line.
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 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab191595 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 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 1/1000 dilution.
Immunoprecipitation - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Immunoprecipitation analysis shows milder lysis buffer is required to get desirable enrichment.

Panel A: RIPA buffer

Panel B: IP lysis buffer (see protocol for details)

HRAS + KRAS was immunoprecipitated from Hepa1-6 (Mouse hepatoma epithelial cell) whole cell lysate 10µg with ab191595 at 1/30 dilution (2ug). Western blot was performed on the immunoprecipitate using ab191595 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 & 4: Hepa1-6 (Mouse hepatoma epithelial cell) whole cell lysate 10µg

Lane 2 & 5: ab191595 IP in Hepa1-6 whole cell lysate

Lane 3 & 6: Rabbit monoclonal IgG (ab172730) instead of ab191595 in Hepa1-6 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes
Immunoprecipitation - Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

KRAS was immunoprecipitated from 1mg of A549 (Human lung carcinoma) whole cell lysate with ab191595 at 1/70 dilution.
Western blot was performed from the immunoprecipitate using ab191595 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: A549 whole cell lysate 10ug (Input).
Lane 2: ab191595 IP in A549 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191595 in A549 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Anti-HRAS + KRAS antibody [EPR18713] (ab191595)

Western blot protocols
Immunoprecipitation protocols
Immunocytochemistry & immunofluorescence protocols
IP lysis buffer preparation

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab191595? Please let us know so that we can cite the reference in this datasheet.

ab191595 has been referenced in 1 publication.

Thu PM et al. Phellodendrine chloride suppresses proliferation of KRAS mutated pancreatic cancer cells through inhibition of nutrients uptake via macropinocytosis. Eur J Pharmacol 850:23-34 (2019). PubMed: 30716311

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Application

Immunoprecipitation

Sample

Human Cell lysate - whole cell (PANC-1 cells)

Total protein in input

250 µg

Immuno-precipitation step

Protein A/G

Specification

PANC-1 cells

Abcam user community

Verified customer

Submitted May 21 2021

Application

Immunoprecipitation

Sample

Mouse Cell lysate - whole cell (MEFs)

Total protein in input

250 µg

Immuno-precipitation step

Protein A/G

Specification

MEFs

Abcam user community

Verified customer

Submitted May 21 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (MEF)

Gel Running Conditions

Reduced Denaturing (12%)

Loading amount

25 µg

Specification

MEF

Blocking step

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

Abcam user community

Verified customer

Submitted Jan 26 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (PANC-1 cells)

Gel Running Conditions

Reduced Denaturing

Loading amount

20 µg

Specification

PANC-1 cells

Blocking step

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

Abcam user community

Verified customer

Submitted Jan 06 2021

Application

ELISA

Sample

Human Recombinant protein (KRAS and HRAS protein)

Specification

KRAS and HRAS protein

Blocking step

Superblock (Thermo Fisher) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Type

Direct

Abcam user community

Verified customer

Submitted Jan 05 2021

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

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

Applications

The Abpromise guarantee

Target

Relevance

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Immunoprecipitation abreview for Anti-HRAS + KRAS antibody [EPR18713]

Immunoprecipitation abreview for Anti-HRAS + KRAS antibody [EPR18713]

Western blot abreview for Anti-HRAS + KRAS antibody [EPR18713]

Western blot abreview for Anti-HRAS + KRAS antibody [EPR18713]

ELISA abreview for Anti-HRAS + KRAS antibody [EPR18713]