Rabbit Anti-HPV16 E7 antibody_Primary Antibody_Antibody_SUNLONG BIOTECH CO., LTD-杭州圣隆生物科技有限公司

Rabbit Anti-HPV16 E7 antibody

E7; HPV16 E7 protein; Human Papilloma Virus; Human papillomavirus type 16 E7; Human papillomavirus type 16 E7; Protein E7; Human Papillomavirus 16 (E7); HPV16 E7; E7 protein (HPV16); VE7_HPV16; HPV16-E7.

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NO.:SL10446R

Clonality:Polyclonal

Immunogen Species:Rabbit

Applications:ELISA IHC-P IHC-F Flow-Cyt ICC IF

concentration:1mg/ml
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Product Name HPV16 E7

Chinese Name 人类乳头状瘤病毒16-E7抗体

Alias E7; HPV16 E7 protein; Human Papilloma Virus; Human papillomavirus type 16 E7; Human papillomavirus type 16 E7; Protein E7; Human Papillomavirus 16 (E7); HPV16 E7; E7 protein (HPV16); VE7_HPV16; HPV16-E7.

literatures
Specific References  (6)     |     SL10446R has been referenced in 6 publications.

[IF=6.852] Yang JH et al. HPV16 E6/E7 upregulate hTERC mRNA and gene amplification levels by relieving the effect of LKB1 on Sp1 phosphorylation in lung cancer cells:. Ther Adv Med Oncol.2020 May 12;12:1758835920917562.  WB ;  Human.

PubMed:32499837

[IF=6.126] Paola Matarrese. et al. Physical Interaction between HPV16E7 and the Actin-Binding Protein Gelsolin Regulates Epithelial-Mesenchymal Transition via HIPPO-YAP Axis. Cancers. 2021 Jan;13(2):353  FC ;  Human.

PubMed:33477952

[IF=3.998] Koji K et al. Evaluation of HPV16 E7 expression in head and neck carcinoma cell lines and clinical specimensSci Rep.2020 Dec 17;10(1):22138.  WB ;  Human.

PubMed:33335126

[IF=3.65] Shao, Jian-Shuang, et al. "HPV16 E6/E7 upregulates HIF-2α and VEGF by inhibiting LKB1 in lung cancer cells." Tumor Biology 39.7 (2017): 1010428317717137.  WB ;  Human.

PubMed:28720067

[IF=3.24] Prabakaran D. S.. et al. Fused toes homolog, a potential molecular regulator of human papillomavirus type 16 E6 and E7 oncoproteins in cervical cancer. PLOS ONE. 2022 Apr;17(4):e0266532  IF ;  Human.

PubMed:10.1371/journal.pone.0266532

[IF=3.08] Liu, Yuan, et al. "Targeting hexokinase 2 inhibition promotes radiosensitization in HPV16 E7-induced cervical cancer and suppresses tumor growth." International Journal of Oncology 50.6 (2017): 2011-2023.  WB ;  Human.

PubMed:28498475

Research Area Tumour

Immunogen Species Rabbit

Clonality Polyclonal

Applications ELISA=1:5000-10000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1ug/Test ICC=1:100-500 IF=1:100-500 （Paraffin sections need antigen repair）
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.

Theoretical molecular weight 11kDa

Cellular localization The nucleus

Form Liquid

Concentration 1mg/ml

immunogen KLH conjugated synthetic peptide derived from human HPV16 E7: 21-98/98

Lsotype IgG

Purification affinity purified by Protein A

Buffer Solution 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.

Storage Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Attention This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

PubMed PubMed

Product Detail Human papilloma viruses (HPVs) can be classified as either high risk or low risk according to their association with cancer. HPV16 and HPV18 are the most common of the high risk group while HPV6 and HPV11 are among the low risk types. Approximately 90% of cervical cancers contain HPV DNA of the high risk types. Mutational analysis have shown that the E6 and E7 genes of the high risk HPVs are necessary and sufficient for HPV transforming function. The specific interactions of the E6 and E7 proteins with p53 and pRB, respectively, correlate with HPV high and low risk classifications. The high risk HPV E7 proteins bind to pRB with a higher affinity than do the low risk HPV proteins, and only the high risk HPV E6 proteins form detectable complexes with p53 in vitro.

Function:
E7 protein has both transforming and trans-activating activities. Disrupts the function of host retinoblastoma protein RB1/pRb, which is a key regulator of the cell cycle. Induces the disassembly of the E2F1 transcription factors from RB1, with subsequent transcriptional activation of E2F1-regulated S-phase genes. Inactivation of the ability of RB1 to arrest the cell cycle is critical for cellular transformation, uncontrolled cellular growth and proliferation induced by viral infection. Stimulation of progression from G1 to S phase allows the virus to efficiently use the cellular DNA replicating machinery to achieve viral genome replication. Interferes with histone deacetylation mediated by HDAC1 and HDAC2, leading to activation of transcription.

Subunit:
Homodimer. Homooligomer. Interaction with host RB1 induces the aberrant dissociation of RB1-E2F1 complex thereby disrupting RB1's activity. Binds to CHD3 through its zinc-finger domain. Forms a complex with CHD3 and HDAC1, thereby altering the action of host histone deacetylation. A similar complex involving E7, CHD3 and HDAC2 might also form. Interacts with E2; this interaction inhibits E7 oncogenic activity.

Similarity:
Belongs to the papillomaviridae E7 protein family.

SWISS:
P03129

Gene ID:
1489079

Database links:

Entrez Gene: 1489079 HPV16

SwissProt: P03129 HPV16

Product Picture

Tissue/cell: Human parotid tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-HPV16 E7 Polyclonal Antibody, Unconjugated(SL10446R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-HPV16 E7 antibody (SL10446R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Black line : Positive blank control (Hela); Negative blank control (RSC96）
Green line : Primary Antibody (Rabbit Anti-HPV16 E7 antibody (SL10446) )
Orange line：Isotype Control Antibody (Rabbit IgG) .
Blue line : Secondary Antibody (Goat anti-rabbit IgG-AF647)
Hela（Positive）and RSC96（Negative control）cells (black) were fixed with 4% PFA for 10min at room temperature, permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HPV16 E7 Antibody(SL10446R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

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Product Name	HPV16 E7
Chinese Name	人类乳头状瘤病毒16-E7抗体
Alias	E7; HPV16 E7 protein; Human Papilloma Virus; Human papillomavirus type 16 E7; Human papillomavirus type 16 E7; Protein E7; Human Papillomavirus 16 (E7); HPV16 E7; E7 protein (HPV16); VE7_HPV16; HPV16-E7.
literatures	Specific References (6) \| SL10446R has been referenced in 6 publications. [IF=6.852] Yang JH et al. HPV16 E6/E7 upregulate hTERC mRNA and gene amplification levels by relieving the effect of LKB1 on Sp1 phosphorylation in lung cancer cells:. Ther Adv Med Oncol.2020 May 12;12:1758835920917562. WB ; Human. PubMed:32499837 [IF=6.126] Paola Matarrese. et al. Physical Interaction between HPV16E7 and the Actin-Binding Protein Gelsolin Regulates Epithelial-Mesenchymal Transition via HIPPO-YAP Axis. Cancers. 2021 Jan;13(2):353 FC ; Human. PubMed:33477952 [IF=3.998] Koji K et al. Evaluation of HPV16 E7 expression in head and neck carcinoma cell lines and clinical specimensSci Rep.2020 Dec 17;10(1):22138. WB ; Human. PubMed:33335126 [IF=3.65] Shao, Jian-Shuang, et al. "HPV16 E6/E7 upregulates HIF-2α and VEGF by inhibiting LKB1 in lung cancer cells." Tumor Biology 39.7 (2017): 1010428317717137. WB ; Human. PubMed:28720067 [IF=3.24] Prabakaran D. S.. et al. Fused toes homolog, a potential molecular regulator of human papillomavirus type 16 E6 and E7 oncoproteins in cervical cancer. PLOS ONE. 2022 Apr;17(4):e0266532 IF ; Human. PubMed:10.1371/journal.pone.0266532 [IF=3.08] Liu, Yuan, et al. "Targeting hexokinase 2 inhibition promotes radiosensitization in HPV16 E7-induced cervical cancer and suppresses tumor growth." International Journal of Oncology 50.6 (2017): 2011-2023. WB ; Human. PubMed:28498475
Research Area	Tumour
Immunogen Species	Rabbit
Clonality	Polyclonal
Applications	ELISA=1:5000-10000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1ug/Test ICC=1:100-500 IF=1:100-500 （Paraffin sections need antigen repair） not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user.
Theoretical molecular weight	11kDa
Cellular localization	The nucleus
Form	Liquid
Concentration	1mg/ml
immunogen	KLH conjugated synthetic peptide derived from human HPV16 E7: 21-98/98
Lsotype	IgG
Purification	affinity purified by Protein A
Buffer Solution	0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Storage	Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Attention	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
PubMed	PubMed
Product Detail	Human papilloma viruses (HPVs) can be classified as either high risk or low risk according to their association with cancer. HPV16 and HPV18 are the most common of the high risk group while HPV6 and HPV11 are among the low risk types. Approximately 90% of cervical cancers contain HPV DNA of the high risk types. Mutational analysis have shown that the E6 and E7 genes of the high risk HPVs are necessary and sufficient for HPV transforming function. The specific interactions of the E6 and E7 proteins with p53 and pRB, respectively, correlate with HPV high and low risk classifications. The high risk HPV E7 proteins bind to pRB with a higher affinity than do the low risk HPV proteins, and only the high risk HPV E6 proteins form detectable complexes with p53 in vitro. Function: E7 protein has both transforming and trans-activating activities. Disrupts the function of host retinoblastoma protein RB1/pRb, which is a key regulator of the cell cycle. Induces the disassembly of the E2F1 transcription factors from RB1, with subsequent transcriptional activation of E2F1-regulated S-phase genes. Inactivation of the ability of RB1 to arrest the cell cycle is critical for cellular transformation, uncontrolled cellular growth and proliferation induced by viral infection. Stimulation of progression from G1 to S phase allows the virus to efficiently use the cellular DNA replicating machinery to achieve viral genome replication. Interferes with histone deacetylation mediated by HDAC1 and HDAC2, leading to activation of transcription. Subunit: Homodimer. Homooligomer. Interaction with host RB1 induces the aberrant dissociation of RB1-E2F1 complex thereby disrupting RB1's activity. Binds to CHD3 through its zinc-finger domain. Forms a complex with CHD3 and HDAC1, thereby altering the action of host histone deacetylation. A similar complex involving E7, CHD3 and HDAC2 might also form. Interacts with E2; this interaction inhibits E7 oncogenic activity. Similarity: Belongs to the papillomaviridae E7 protein family. SWISS: P03129 Gene ID: 1489079 Database links: Entrez Gene: 1489079 HPV16 SwissProt: P03129 HPV16
Product Picture	Tissue/cell: Human parotid tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min; Incubation: Anti-HPV16 E7 Polyclonal Antibody, Unconjugated(SL10446R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (Human uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (Human uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (SL10446R-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Blank control: Hela. Primary Antibody (green line): Rabbit Anti-HPV16 E7 antibody (SL10446R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PE Dilution: 1μg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed. Black line : Positive blank control (Hela); Negative blank control (RSC96） Green line : Primary Antibody (Rabbit Anti-HPV16 E7 antibody (SL10446) ) Orange line：Isotype Control Antibody (Rabbit IgG) . Blue line : Secondary Antibody (Goat anti-rabbit IgG-AF647) Hela（Positive）and RSC96（Negative control）cells (black) were fixed with 4% PFA for 10min at room temperature, permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HPV16 E7 Antibody(SL10446R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).