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[KO Validated] YAP1 Rabbit mAb (A21216)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of lysates from wild type (WT) and YAP1 knockout (KO) HeLa cells using [KO Validated] YAP1 Rabbit mAb (A21216) at 1:20000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Western blot analysis of lysates from NIH/3T3 cells using [KO Validated] YAP1 Rabbit mAb (A21216) at 1:20000 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

ABclonal:Immunohistochemistry - [KO Validated] YAP1 Rabbit mAb (A21216)

Immunohistochemistry analysis of paraffin-embedded human breast cancer tissue using [KO Validated] YAP1 Rabbit mAb (A21216) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded human thyroid cancer tissue using [KO Validated] YAP1 Rabbit mAb (A21216) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded human tonsil tissue using [KO Validated] YAP1 Rabbit mAb (A21216) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded mouse spleen tissue using [KO Validated] YAP1 Rabbit mAb (A21216) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded rat spleen tissue using [KO Validated] YAP1 Rabbit mAb (A21216) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - [KO Validated] YAP1 Rabbit mAb (A21216)

Confocal imaging of HeLa cells using [KO Validated] YAP1 Rabbit mAb (A21216, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of C2C12 cells using [KO Validated] YAP1 Rabbit mAb (A21216, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Immunoprecipitation - [KO Validated] YAP1 Rabbit mAb (A21216)

Immunoprecipitation analysis of 300 μg extracts of HeLa cells using 3 μg YAP1 antibody (A21216). Western blot was performed from the immunoprecipitate using [KO Validated] YAP1 Rabbit mAb (A21216) at a dilution of1:2000.

ABclonal:Chromatin Immunoprecipitation - [KO Validated] YAP1 Rabbit mAb (A21216)

Chromatin immunoprecipitation analysis of extracts of HeLa cells, using [KO Validated] YAP1 Rabbit mAb (A21216) and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.

Overview

Product name	[KO Validated] YAP1 Rabbit mAb
Catalog No.	A21216
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC53479

Background

This gene encodes a downstream nuclear effector of the Hippo signaling pathway which is involved in development, growth, repair, and homeostasis. This gene is known to play a role in the development and progression of multiple cancers as a transcriptional regulator of this signaling pathway and may function as a potential target for cancer treatment. Alternative splicing results in multiple transcript variants encoding different isoforms.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 155-504 of human YAP1 (NP_001123617.1).
Sequence	PTAQHLRQSSFEIPDDVPLPAGWEMAKTSSGQRYFLNHIDQTTTWQDPRKAMLSQMNVTAPTSPPVQQNMMNSASGPLPDGWEQAMTQDGEIYYINHKNKTTSWLDPRLDPRFAMNQRISQSAPVKQPPPLAPQSPQGGVMGGSNSNQQQQMRLQQLQMEKERLRLKQQELLRQAMRNINPSTANSPKCQELALRSQLPTLEQDGGTQNPVSSPGMSQELRTMTTNSSDPFLNSGTYHSRDESTDSGLSMSSYSVPRTPDDFLNSVDEMDTGDTINQSTLPSQQNRFPDYLEAIPGTNVDLGTLEGDGMNIEGEELMPSLQEALSSDILNDMESVLAATKLDKESFLTWL
Gene ID
Swiss Prot
Synonyms	YAP; YKI; COB1; YAP2; YAP-1; YAP65; P1
Calculated MW	54kDa
Observed MW	70kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:20000 - 1:80000 IHC-P 1:200 - 1:800 IF/ICC 1:200 - 1:800 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements. ChIP 5μg antibody for 10μg-15μg of Chromatin
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence Immunoprecipitation
Positive samples	HeLa, NIH/3T3
Cellular location	Cytoplasm, Nucleus.
Customer validation	WB(Homo sapiens, Other) IF(Mus musculus) IHC(Mus musculus) WB(Mus musculus)