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[KO Validated] Ki67 Rabbit PolymAb® (A26755PM)

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 Ki67 knockout (KO) HeLa cells using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at 1:1000 dilution incubated overnight at 4℃. HeLa cells and Ki67 knockout (KO) HeLa cells were treated by Nocodazole (1 μg/mL) at 37℃ for 24 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of lysates from RAW264.7 cells using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at 1:1000 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: 5s.

ABclonal:Immunohistochemistry - [KO Validated] Ki67 Rabbit PolymAb® (A26755PM)

Immunohistochemistry analysis of paraffin-embedded Human esophagus tissue using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at a dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at a dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at a dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at a dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat colon tissue using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at a dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM) at a dilution of 1:1000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.

ABclonal:Immunofluorescence - [KO Validated] Ki67 Rabbit PolymAb® (A26755PM)

Confocal imaging of C2C12 cells using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM, 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 A-431 cells using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM, 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 HeLa cells using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM, 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 PC-12 cells using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM, 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 paraffin-embedded Mouse spleen tissue using [KO Validated] Ki67 Rabbit PolymAb® (A26755PM, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IF staining. Objective: 40x.

Overview

Product name	[KO Validated] Ki67 Rabbit PolymAb®
Catalog No.	A26755PM
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

Enables protein C-terminus binding activity. Involved in regulation of chromosome segregation and regulation of mitotic nuclear division. Located in chromosome; nuclear body; and nucleolus. Colocalizes with condensed chromosome. Implicated in Crohn's disease; breast cancer; human immunodeficiency virus infectious disease; and pancreatic cancer. Biomarker of several diseases, including Barrett's esophagus; autoimmune disease of musculoskeletal system (multiple); endocrine gland cancer (multiple); gastrointestinal system cancer (multiple); and interstitial cystitis.

Immunogen information

Immunogen	A synthetic peptide corresponding to a sequence within amino acids 2271-2370 of human Ki67 (NP_002408.3).
Sequence	PAGGDEKDMKAFMGTPVQKLDLPGNLPGSKRWPQTPKEKAQALEDLAGFKELFQTPGTDKPTTDEKTTKIACKSPQPDPVDTPASTKQRPKRNLRKADVE
Gene ID
Swiss Prot
Synonyms	KIA; MIB-; MIB-1; PPP1R105
Calculated MW	350kDa/359kDa
Observed MW	359kDa/359kDa以下

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 1:1000 - 1:4000 IF/ICC 1:200 - 1:2000 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 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	HeLa, RAW264.7
Cellular location	Chromosome, Nucleus, nucleolus.