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YB-1/YBX1 Rabbit mAb (A3534)

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 HeLa cells, using YB-1/YBX1 Rabbit mAb (A3534) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1s.

Western blot analysis of various lysates using YB-1/YBX1 Rabbit mAb (A3534) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

ABclonal:Immunohistochemistry - YB-1/YBX1 Rabbit mAb (A3534)

Immunohistochemistry analysis of paraffin-embedded Human colon using YB-1/YBX1 Rabbit mAb (A3534) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human thyroid cancer using YB-1/YBX1 Rabbit mAb (A3534) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse kidney using YB-1/YBX1 Rabbit mAb (A3534) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat colon using YB-1/YBX1 Rabbit mAb (A3534) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat kidney using YB-1/YBX1 Rabbit mAb (A3534) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - YB-1/YBX1 Rabbit mAb (A3534)

Confocal imaging of U-2 OS cells using YB-1/YBX1 Rabbit mAb (A3534, 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 NIH/3T3 cells using YB-1/YBX1 Rabbit mAb (A3534, 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 C6 cells using YB-1/YBX1 Rabbit mAb (A3534, 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 - YB-1/YBX1 Rabbit mAb (A3534)

Immunoprecipitation of YB-1/YBX1 from 200 µg extracts of HeLa cells was performed using 0.5 µg of YB-1/YBX1 Rabbit mAb (A3534). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using YB-1/YBX1 Rabbit mAb (A3534) at a dilution of 1:1000.

Overview

Product name	YB-1/YBX1 Rabbit mAb
Catalog No.	A3534
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC0797

Background

This gene encodes a highly conserved cold shock domain protein that has broad nucleic acid binding properties. The encoded protein functions as both a DNA and RNA binding protein and has been implicated in numerous cellular processes including regulation of transcription and translation, pre-mRNA splicing, DNA reparation and mRNA packaging. This protein is also a component of messenger ribonucleoprotein (mRNP) complexes and may have a role in microRNA processing. This protein can be secreted through non-classical pathways and functions as an extracellular mitogen. Aberrant expression of the gene is associated with cancer proliferation in numerous tissues. This gene may be a prognostic marker for poor outcome and drug resistance in certain cancers. Alternate splicing results in multiple transcript variants. Pseudogenes of this gene are found on multiple chromosomes.

Immunogen information

Immunogen	A synthetic peptide corresponding to a sequence within amino acids 225-324 of human YB-1/YBX1 (P67809).
Sequence	GAGEQGRPVRQNMYRGYRPRFRRGPPRQRQPREDGNEEDKENQGDETQGQQPPQRRYRRNFNYRRRRPENPKPQDGKETKAADPPAENSSAPEAEQGGAE
Gene ID
Swiss Prot
Synonyms	YB1; BP-8; CSDB; DBPB; YB-1; CBF-A; CSDA2; EFI-A; NSEP1; NSEP-1; MDR-NF1; YB-1/YBX1
Calculated MW	36kDa
Observed MW	49kDa/

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 1:1000 - 1:6000 IHC-P 1:200 - 1:2000 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.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.02% sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence Immunoprecipitation
Positive samples	Mouse testis, Rat heart, HeLa, HeLa
Cellular location	Cytoplasm, Cytoplasmic granule, Nucleus, Secreted.
Customer validation	WB(Homo sapiens)