Knockout validation as standard antibody validation

One of the most accepted and trusted validation processes for antibody specificity is through knockout validation (Madhusoodanan, 2014; Rhodes & Trimmers, 2008). In a knockout validation, antibody specificity is confirmed by testing the antibody of interest on a knockout sample or cell line which doesn't express the target protein.  In a knockout cell line, the antibody target is not present as the gene encoding the protein of interest is eliminated or "knocked out". This data is compared side-by-side against a normal cell line (or wild type) and if the antibody is truly specific, the antibody should have no detection in the knockout cell line, but instead a specific signal in the normal cell line. Knockout validations offers a true negative control to confirm the antibody specificity to the protein of interest (Bordeaux, et al., 2010).

See all the knockout validation data below

ABCF2 Knockout 293T Cell Lysate [L0046099004]

Gene Symbol: ABCF2

NCBI Gene ID: 10061

Other Namesp: ABC28; HUSSY18; HUSSY-18; EST133090

Parental Cell Line: 293T

Application: WB; KO Validation

L0046099004-01

Anti-ABCF2  Antibody from OriGene
SKU: TA811105

Lanes WT: Wild-type HeLa cell lysate (10 µg)
Lanes KO: CDK4 knockout HeLa cell lysate (10 µg)

Predicted band size : 80 kDa
Observed band size : 80 kDa

Equivalent amounts of cell lysates (10 µg per lane) of wild-type 293T cells and ABCF2-Knockout 293T cells (KO, Cat# L0046099004) were separated by SDS-PAGE and immunoblotted with anti-ABCF2 monoclonal antibody from OriGene (Cat# TA811105),(1:500). Then the blotted membrane was stripped and reprobed with anti-b-actin antibody as a loading control.

L0046099004-02

Anti-ABCF2  Antibody from OriGene
SKU: TA811212

Lanes WT: Wild-type HeLa cell lysate (10 µg)
Lanes KO: CDK4 knockout HeLa cell lysate (10 µg)

Predicted band size : 80 kDa
Observed band size : 80 kDa

Equivalent amounts of cell lysates (10 µg per lane) of wild-type 293T cells and ABCF2-Knockout 293T cells (KO, Cat# L0046099004) were separated by SDS-PAGE and immunoblotted with anti-ABCF2 monoclonal antibody from OriGene (Cat# TA811212),(1:500). Then the blotted membrane was stripped and reprobed with anti-b-actin antibody as a loading control.

 

 

ADAM17 Knockout HeLa Cell Lysate [L0015573702]
ADAM17 Knockout HeLa Cell Lysate [L0015573702]

Gene Symbol: ADAM17

NCBI Gene ID: 6868

Other Namesp: CSVP; TACE; NISBD; ADAM18; CD156B; NISBD1

Parental Cell Line: Hela

Application: WB; KO Validation

 

L0015573702

Anti-ADAM17  Antibody from GeneTex
SKU: GTX101358

 

 

 

AHSA1 Knockout HeLa Cell Lysate [L0019478704]

Gene Symbol: AHSA1

NCBI Gene ID: 10598

Other Namesp: p38; AHA1; hAha1; C14orf3

Parental Cell Line: HeLa

Application: WB; KO Validation

L0019478704-01-N

Anti-AHSA1  Antibody from OriGene
SKU: TA808924

Equivalent amounts of cell lysates (10 ug per lane) of wild-type Hela cells (WT, Cat# LC810HELA) and AHSA1-Knockout Hela cells (KO, Cat# L0019478704) were separated by SDS-PAGE and immunoblotted with anti-AHSA1 monoclonal antibody TA808924. Then the blotted membrane was stripped and reprobed with anti-HSP90AB1 antibody as a loading control.(1:500)

L0019478704

Anti-AHSA1  Antibody from GeneTex
SKU: GTX102312

 

 

AKR1B1 Knockout Hela Cell Lysate [L0014159205]

Gene Symbol: AKR1B1

NCBI Gene ID: 231

Other Namesp: AR; ADR; ALR2; ALDR1

Parental Cell Line: 293T

Application: WB; KO Validation

GTX113381 40128 20170427 WB KO

Anti-AKR1B1  Antibody from GeneTex
SKU: GTX113381

Wild-type (WT) and AKR1B1 knockout (KO) HeLa cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with AKR1B1 antibody [N1C3] (GTX113381) diluted at 1:2500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

 

 

ANXA1 Knockout HeLa Cell Lysate [L0018809501]

 

Gene Symbol: ANXA1

NCBI Gene ID: 301

Other Namesp: ANX1; LPC1

Parental Cell Line: HeLa

Application: WB; KO Validation

L0018809501-01

Anti-ANXA1 Antibody from OriGene
SKU: TA500967

Equivalent amounts of cell lysates (10 ug per lane) of wild-type HeLa cells (WT, Cat# LC810HELA) and ANXA1-Knockout hela cells (KO,Cat# L0018809501) were separated by SDS-PAGE and immunoblotted with anti-ANXA1 monoclonal antibody TA500967. Then the blotted membrane was stripped and reprobed with anti-HSP90AB1 antibody as a loading control.(1:500)

 

 

 

CAST Knockout HeLa Cell Lysate [L0013017003]
 

Gene Symbol: CAST

NCBI Gene ID: 831

Other Namesp: BS-17; PLACK

Parental Cell Line: HeLa

Application: WB; KO Validation

 

L0013017003-01-N

Anti-CAST  Antibody from OriGene
SKU: TA500967

Equivalent amounts of cell lysates (10 ug per lane) of wild-type Hela cells (WT, Cat# LC810HELA) and CAST-Knockout Hela cells (KO, Cat# L0013017003) were separated by SDS-PAGE and immunoblotted with anti-CAST monoclonal antibody TA504351,(1:500). Then the blotted membrane was stripped and reprobed with anti-HSP90AB1 antibody as a loading control.

 

 

 

The data is continues to expending……

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