Validation Data Gallery

Filter: AllWBIPIHCIF/ICC

WB analysis using 15194-1-AP
Various lysates were subjected to SDS PAGE followed by western blot with 15194-1-AP (DHFR antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.
WB analysis of HeLa using 15194-1-AP
WB result of DHFR antibody (15194-1-AP; 1:5000; incubated at room temperature for 1.5 hours) with sh-Control and sh-DHFR transfected HeLa cells.
WB analysis of HeLa using 15194-1-AP
HeLa cells were subjected to SDS PAGE followed by western blot with 15194-1-AP (DHFR antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of Raji using 15194-1-AP
Raji cells were subjected to SDS PAGE followed by western blot with 15194-1-AP (DHFR antibody) at dilution of 1:300 incubated at room temperature for 1.5 hours.
WB analysis of Jurkat using 15194-1-AP
Jurkat cells were subjected to SDS PAGE followed by western blot with 15194-1-AP (DHFR antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
WB analysis of K-562 using 15194-1-AP
K-562 cells were subjected to SDS PAGE followed by western blot with 15194-1-AP (DHFR antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.
IP experiment of Raji using 15194-1-AP
IP result of anti-DHFR (IP:15194-1-AP, 3ug; Detection:15194-1-AP 1:500) with Raji cells lysate 2000ug.
IHC staining of human liver cancer using 15194-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 15194-1-AP (DHFR antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human liver cancer using 15194-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 15194-1-AP (DHFR antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of human breast cancer using 15194-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 15194-1-AP (DHFR Antibody) at dilution of 1:50 (under 10x lens).
IHC staining of human breast cancer using 15194-1-AP
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 15194-1-AP (DHFR Antibody) at dilution of 1:50 (under 40x lens).
IHC staining of human liver cancer using 15194-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 15194-1-AP (DHFR Antibody) at dilution of 1:50 (under 10x lens).
IHC staining of human liver cancer using 15194-1-AP
Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 15194-1-AP (DHFR Antibody) at dilution of 1:50 (under 40x lens).
IF Staining of HeLa using 15194-1-AP
Immunofluorescent analysis of HeLa cells using 15194-1-AP (DHFR antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

Featured Product
KD/KO Validated

DHFR Polyclonal antibody

DHFR Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 15194-1-AP

Publications(26)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, IP, ELISA

Synonyms

DHFRP1, EC:1.5.1.3

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (26)
Protocols

データシート［PDF］を印刷

MSDS

Tested Applications

Positive WB detected in	HEK-293 cells, HeLa cells, Jurkat cells, K-562 cells, Raji cells, mouse kidney tissue, mouse liver tissue, rat kidney tissue
Positive IP detected in	Raji cells
Positive IHC detected in	human liver cancer tissue, human breast 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

Application	Dilution
Western Blot (WB)	WB : 1:1000-1:6000
Immunoprecipitation (IP)	IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
Immunohistochemistry (IHC)	IHC : 1:50-1:500
Immunofluorescence (IF)/ICC	IF/ICC : 1:20-1:200
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

KD/KO	See 5 publications below
WB	See 21 publications below
IF	See 3 publications below

Product Information

15194-1-AP targets DHFR in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human, mouse, rat samples.

Tested Reactivity	human, mouse, rat
Cited Reactivity	human, mouse, zebrafish
Host / Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	CatNo: Ag7331 Product name: Recombinant human DHFR protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 1-187 aa of BC000192 Sequence: MVGSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND 相同性解析による交差性が予測される生物種
Full Name	dihydrofolate reductase
Calculated molecular weight	21 kDa
Observed molecular weight	21 kDa
GenBank accession number	BC000192
Gene Symbol	DHFR
Gene ID (NCBI)	1719
RRID	AB_2091587
Conjugate	Unconjugated
Form
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	P00374
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

DHFR (dihydrofolate reductase) catalyzes the subsequent NADPH-dependent reduction of the H2folate produced by TS to regenerate the tetrahydrofolate (H4folate) necessary for continued dTMP synthesis and transfer of 1-carbon units (PMID:3461439). It belongs to the dihydrofolate reductase family and can exist as a homodimer (PMID:2248959). DHFR is widely expressed in fetal and adult human brain and whole blood, expression is higher in adult brain than fetal brain (PMID:21310276).

Protocols

Product Specific Protocols
IF protocol for DHFR antibody 15194-1-AP	Download protocol
IHC protocol for DHFR antibody 15194-1-AP	Download protocol
IP protocol for DHFR antibody 15194-1-AP	Download protocol
WB protocol for DHFR antibody 15194-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

Species	Application	Title
human	WB	Nat Cancer ENO1 suppresses cancer cell ferroptosis by degrading the mRNA of iron regulatory protein 1. Authors - Tong Zhang View Article
		Nat Commun Coupling cellular drug-target engagement to downstream pharmacology with CeTEAM Authors - Nicholas C K Valerie View Article
human	WB	Mol Cell mTORC1 stimulates cell growth through SAM synthesis and m6A mRNA-dependent control of protein synthesis. Authors - Elodie Villa View Article
		Mol Cell Accumulation of succinate suppresses de novo purine synthesis through succinylation-mediated control of the mitochondrial folate cycle. Authors - Mushtaq A. Nengroo View Article
human	WB	Genes Dis CYP3A5 unexpectedly regulates glucose metabolism through the AKT-TXNIP-GLUT1 axis in pancreatic cancer Authors - Ming Shao View Article
human	WB,IF	Cell Death Dis Metformin sensitises hepatocarcinoma cells to methotrexate by targeting dihydrofolate reductase. Authors - Yinghui Wang KD Validated View Article

Validation Data Gallery

WB analysis using 15194-1-AP

WB analysis of HeLa using 15194-1-AP

WB analysis of HeLa using 15194-1-AP

WB analysis of Raji using 15194-1-AP

WB analysis of Jurkat using 15194-1-AP

WB analysis of K-562 using 15194-1-AP

IP experiment of Raji using 15194-1-AP

IHC staining of human liver cancer using 15194-1-AP

IHC staining of human liver cancer using 15194-1-AP

IHC staining of human breast cancer using 15194-1-AP

IHC staining of human breast cancer using 15194-1-AP

IHC staining of human liver cancer using 15194-1-AP

IHC staining of human liver cancer using 15194-1-AP

IF Staining of HeLa using 15194-1-AP

DHFR Polyclonal antibody

DHFR Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

ENO1 suppresses cancer cell ferroptosis by degrading the mRNA of iron regulatory protein 1.

Coupling cellular drug-target engagement to downstream pharmacology with CeTEAM

mTORC1 stimulates cell growth through SAM synthesis and m6A mRNA-dependent control of protein synthesis.

Accumulation of succinate suppresses de novo purine synthesis through succinylation-mediated control of the mitochondrial folate cycle.

CYP3A5 unexpectedly regulates glucose metabolism through the AKT-TXNIP-GLUT1 axis in pancreatic cancer

Metformin sensitises hepatocarcinoma cells to methotrexate by targeting dihydrofolate reductase.

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