KRAS-4B-specific Polyclonal antibody

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  WB analysis of HeLa using 16155-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 16155-1-AP (KRAS-2B-specific antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis using 16155-1-AP

  Recombinant proteins(ag27444 (6*His tag-KRAS-2A), ag21645(6*His tag-KRAS-2B), ag29746(6*His tag-NRAS), ag23265(GST-HRAS))were subjected to SDS PAGE followed by western blot with 16155-1-AP (KRAS-2B-specific antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 16155-1-AP

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 16155-1-AP (KRAS-2B-specific antibody) at dilution of 1:1200 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 16155-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 16155-1-AP (KRAS-2B-specific antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 16155-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 16155-1-AP (KRAS-2B-specific antibody) at dilution of 1:1500 incubated at room temperature for 1.5 hours.

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  IHC staining of human colon cancer using 16155-1-AP

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 16155-1-AP (KRAS-2B-specific antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon cancer using 16155-1-AP

  Immunohistochemical analysis of paraffin-embedded human colon cancer tissue slide using 16155-1-AP (KRAS-2B-specific antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 16155-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using KRAS-2B-specific antibody (16155-1-AP) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).

• Featured Product
• KD/KO Validated

KRAS-4B-specific Polyclonal Antibody for WB, IHC, IF/ICC, ELISA

Cat No. 16155-1-AP

Publications(6)

Reviews (1)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

KRAS-4B, KRAS, pan RAS, C K RAS, c Ki ras

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, MCF-7 cells, mouse brain tissue, Recombinant protein
• Positive IHC detected in: human colon cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 4 publications below

Product Information

16155-1-AP targets KRAS-4B-specific in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog
• Calculated Molecular Weight: 21 kDa
• Observed Molecular Weight: 21 kDa
• GenBank Accession Number: NM_004985
• Gene Symbol: KRAS
• Gene ID (NCBI): 3845
• RRID: AB_2134119
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P01116
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

RAS gene family encodes very similar 21KD proteins and are homologs of rodent sarcoma virus genes that have transforming abilities. KRAS has two isoforms, 4A and 4B. This antibody is specific for KRAS-4B, which is also named as KRAS-2B.

Publications

Species	Application	Title
human		Nat Biotechnol; Conformation-locking antibodies for the discovery and characterization of KRAS inhibitors.; Authors - Christopher W Davies
human		Nat Biomed Eng; Tracking the expression of therapeutic protein targets in rare cells by antibody-mediated nanoparticle labelling and magnetic sorting.; Authors - Mahmoud Labib
human	WB	Cancer Res; Targeting eIF4A-Dependent Translation of KRAS Signaling Molecules.; Authors - Kamini Singh
human	WB	Cell Chem Biol; Targeting Endogenous K-RAS for Degradation through the Affinity-Directed Protein Missile System.; Authors - Sascha Röth
human	WB	Sci Signal; Evaluation of the selectivity and sensitivity of isoform- and mutation-specific RAS antibodies.; Authors - Andrew M Waters
human	WB	Life Sci Alliance; A new ferrocene derivative blocks K-Ras localization and function by oxidative modification at His95; Authors - Kristen M Rehl

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Christopher (Verified Customer) (12-14-2018)

Antibody is very specific but also quite weak. Maybe you can increase the primary antibody dilution to 1:500 to increase signal. There is no unspecific recognition of H-Ras or N-Ras (see picture). Also endogenous Ras only exhibits one band and not two which is a hint for the recognition of endogenous N-Ras next to K-Ras.In immunofluorescence experiments antibody works awesome at a dilution of 1:100 for pancreatic cancer cell lines. In total I'm very satisfied, but you have to use more antibody than written in the data sheet.

• Applications: Western Blot, Immunofluorescence, Immunoprecipitation,
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: pancreatic cancer cells (PANC-1) and HEK293 overexpressing EGFP-tagged K-Ras, H-Ras and N-Ras

KRAS-4B-specific Antibody Western Blot,Immunofluorescence,Immunoprecipitation, validation (1:1000 dilution) in pancreatic cancer cells (PANC-1) and HEK293 overexpressing EGFP-tagged K-Ras, H-Ras and N-Ras (Cat no:16155-1-AP)