KRAS-2A-specific Polyclonal antibody

KRAS-2A-specific Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

3

Cat no : 16156-1-AP

Synonyms

KRAS-2A, KRAS, KRAS4A, KRAS4B, pan RAS

---

Validation Data Gallery

• 

  WB analysis of NIH/3T3 using 16156-1-AP

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 16156-1-AP (KRAS-2A-specific antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 16156-1-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.

• 

  IHC staining of human stomach using 16156-1-AP

  Immunohistochemical analysis of paraffin-embedded human stomach tissue slide using 16156-1-AP (KRAS-2A-specific antibody) at dilution of 1:200 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 16156-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using KRAS-2A-specific antibody (16156-1-AP) at dilution of 1:400 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).

• 

  FC experiment of HeLa using 16156-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human KRAS-2A-specific (16156-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP, Clone: ) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

---

Tested Applications

• Positive WB detected in: NIH/3T3 cells
• Positive IHC detected in: human stomach 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
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:1000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

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 3 publications below

Product Information

16156-1-AP targets KRAS-2A-specific in WB, IHC, IF/ICC, FC (Intra), 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: 21-24 kDa
• GenBank Accession Number: NM_033360
• Gene Symbol: KRAS
• Gene ID (NCBI): 3845
• RRID: AB_2234477
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• 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 (derived from 'RAt Sarcoma' virus) gene family includes KRAS, HRAS, and NRAS. These genes encode very similar 21 kDa proteins and are homologs of rodent sarcoma virus genes that have transforming abilities. KRAS has two isoforms, 2A and 2B. This antibody is specific for KRAS-2A, which also named as KRAS-4A .

Publications

Species	Application	Title
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