LBIS® 血蓝蛋白(KLH)(T细胞依赖性抗原) 大鼠免疫球蛋白G(IgG)ELISA试剂盒 LBIS® KLH(TDAR) Rat-IgG ELISA Kit

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

LBIS® 血蓝蛋白(KLH)(T细胞依赖性抗原)  大鼠免疫球蛋白G(IgG)ELISA试剂盒                              LBIS® KLH(TDAR) Rat-IgG ELISA Kit血蓝蛋白(KLH)(T细胞依赖性抗原)

大鼠免疫球蛋白G(IgG)ELISA试剂盒

 


  药品的免疫毒性实验相关方针ICH S8中推荐在无特定免疫毒性标靶情况下进行的T细胞依赖性抗体生产实验(TDAR、T cell Dependent Antibody Reaction)。TDAR被称为T细胞依赖性抗原。例如:通过KLH(Keyhole limpet hemocyanin)的投放对一次抗原刺激而生产IgM性状抗体。接下来进行二次抗原刺激后通过类别转换作用可观察到产生IgG性状抗体。由于本试剂盒可简便测定大鼠血液中IgM性状的抗KLH浓度,所以十分符合上文所述的目的。可与大鼠抗KLH-IgG检测试剂盒配套使用。


◆特点

LBIS® 血蓝蛋白(KLH)(T细胞依赖性抗原)  大鼠免疫球蛋白G(IgG)ELISA试剂盒                              LBIS® KLH(TDAR) Rat-IgG ELISA Kit


● 测定时间短(总反应时间:2小时20分钟)

● 微量样本即可测定。

● 使用无害的防腐剂。

● 全部试剂为溶液即用类型。

● 高测定精度和高重复性。

 

试剂盒组成


组成品

状态

包装

KLH包被96孔板

清洗后使用

96 wells(8×12)/1个

抗KLH大鼠IgG标准溶液(300 ng/mL)

稀释后使用

200 μL/1瓶

缓冲液

直接使用

100 mL/1瓶

HRP结合抗大鼠IgG抗体

稀释后使用

100 μL/1瓶

显色液(TMB)

直接使用

12 mL/1瓶

终止液(1M H2SO4

※小心轻放

直接使用

12 mL/1瓶

浓缩清洗液(10×)

稀释后使用

100 mL/1瓶

孔板密封膜

3个

产品说明书

1本

 


◆物种交叉性


3000 ng/mL时数据—:无交叉性


动物种类

对象物质

反应性及反应率(%)

大鼠

IgG

100

IgM

IgA

IgE

小鼠

IgG

IgM

IgE

 


◆样本信息


● 大鼠血清·血浆

● 50 μL/well(稀释样本)

※ 用附带的缓冲液将样品稀释至标准曲线范围内。

※ 为了避免非特异反应发生,请将样本稀释500倍以上。

 


◆测定范围


0.47~30 ng/mL(标准曲线范围)

 


实验数据


精密度实验(实验内变化)


样本

A

B

1

11.2

3.93

2

11.0

3.74

3

11.3

3.83

4

11.1

3.70

5

11.5

3.99

Mean

11.2

3.84

SD

0.190

0.121

CV(%)

1.7

3.2

单位:ng/mL



重复性实验(实验间变化)


测定日/样本

E

F

G

0天

15.3

3.84

0.969

1天

15.3

3.84

0.954

2天

15.4

3.83

0.958

3天

15.2

3.82

0.976

Mean

15.3

3.83

0.965

SD

0.0615

0.0067

0.0102

CV(%)

0.40

0.18

1.1

单位:ng/mL,n=4



添加回收实验


样本C


添加量

实测值

回收量

回收率(%)

0.00

1.18

0.466

1.65

0.470

101

1.40

2.53

1.35

96.4

2.33

3.48

2.30

98.7

单位:ng/mL,n=2


样本D


添加量

实测值

回收量

回收率(%)

0.00

12.5

4.90

17.2

4.70

95.9

10.7

23.1

10.6

99.1

15.4

27.2

14.7

95.5

单位:ng/mL,n=2



稀释直线性实验


2个血清样本连续用稀释缓冲液稀释3个梯度测定结果,直线回归值R2=0.9992~0.9998

产品编号 产品名称 产品规格 产品等级
636-13759 LBIS® KLH(TDAR) Rat-IgG ELISA Kit 
血蓝蛋白(KLH)(T细胞依赖性抗原)大鼠免疫球蛋白G(IgG) ELISA试剂盒
96 tests

IdeZ Protease (IgG-specific) |NEB酶试剂 New England Biolabs

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

产品信息

IdeZ Protease (IgG-specific) |
 
IdeZ Protease (IgG-specific) is a recombinant antibody specific protease that recognizes all human, sheep, monkey, and rabbit IgG subclasses, specifically cleaving at a single recognition site below the hinge region, yielding a homogenous pool of F(ab´)2 and Fc fragments. IdeZ Protease more effectively cleaves murine IgG2a than IdeS.

IdeZ Protease (IgG-specific) |
Digestion of lgG with IdeZ Protease (lgG-specific), followed by denaturation.

产品来源

Cloned from Streptococcus equi subspecies zooepidemicus and expressed in E. coli. 

产品类别:
Proteases Products,
Proteome Analysis Products

应用:
Proteomics

  • 产品组分信息

    本产品提供以下试剂或组分:

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P0770S     -20    
        IdeZ Protease (IgG-specific) P0770SVIAL -20 1 x 0.05 ml 80,000 units/ml
        GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to cleave > 95% of 1 µg of human IgG, in 15 minutes at 37°C in a total reaction volume of 10 µl.

    反应条件

    1X GlycoBuffer 2
    Incubate at 37°C

    1X GlycoBuffer 2
    50 mM Sodium Phosphate
    (pH 7.5 @ 25°C)

    贮存溶液

    20 mM Tris-HCl
    50 mM NaCl
    1 mM EDTA
    pH 7.5 @ 25°C

    热失活

    65°C for 10 minutes

    分子量

    理论上的: 35578 daltons

    单位活性检测条件

    Two fold dilutions of IdeZ Protease (IgG-specific) are incubated with 1 μg of human IgG and 1X GlycoBuffer 2 in a 10 µl reaction. The reaction mix is incubated for 15 minutes at 37°C. Separation of reaction products are visualized by SDS-PAGE.

    存储注意事项

    • Avoid repeated freeze/thaw cycles.

  • 相关产品

    相关产品

    • Rapid 快速 PNGase F
    • Endo S
    • Trypsin-ultra™, Mass Spectrometry Grade
    • p6043-rapid-pngase-f-antibody-standard
    • p0711-rapid-pngase-f-non-reducing-format

  • 注意事项

    1. Store at -20°C for up to two years.
    2. IdeZ Protease efficiently cleaves human, humanized, chimeric, sheep,rabbit and monkey IgG as well as mouse IgG2a and IgG3. IdeZ Proteasewill also cleave Fc-fusion proteins, such as Enbrel.
    3. IdeZ Protease does not cleave mouse IgG1 or IgG2b, rat, porcine, bovineor goat IgG. It also does not cleave non-IgG isotypes including IgA,IgM, IgD and IgE.

操作说明、说明书 & 用法

  • 操作说明

    1. Reaction Conditions for IdeZ Protease (IgG-specific) (P0770)
    2. Reaction conditions for Simultaneous Digestion of IgG with IdeZ Protease (IgG-specific) and PNGase F (fragmentation and deglycosylation) (P0770)

FAQs & 问题解决指南

  • FAQs

    1. What is the advantage of IdeZ over IdeS?
    2. Is IdeZ Protease active in Phosphate Buffered Saline (PBS)?
    3. Can protein A magnetic beads (NEB# S1425S) be used to create Fc and Fab fragment pools after cleavage with IdeZ Protease?
    4. Can IdeZ Protease cleave IgGs other than human?
    5. Can IdeZ Protease cleave Ig isotypes other than IgG?
    6. Can PNGase F be used together with IdeZ Protease under native conditions to deglycosylate the Fc portion of an antibody?
    7. What is the cleavage site for IdeZ Protease (IgG-specific)?

LBIS® 小鼠卵清蛋白特异性免疫球蛋白G1(OVA-IgG1)ELISA试剂盒 LBIS® OVA-IgG1 Mouse

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

LBIS® 小鼠卵清蛋白特异性免疫球蛋白G1(OVA-IgG1)ELISA试剂盒                              LBIS® OVA-IgG1 MouseLBIS® 小鼠卵清蛋白特异性免疫球蛋白G1(OVA-IgG1)ELISA试剂盒

 


  IgG是血清中免疫球蛋白中含量最高的二次免疫应答主要抗体。占据三分之二IgG的是具有γ1的H链,分子量146 kDa的IgG1。以OVA(卵清蛋白)作为特殊化抗原,通过简化测定抗OVA- IgG1抗体值来阐明小鼠免疫系统作用机理。

  小鼠卵清蛋白特异性免疫球蛋白G1(OVA-IgG1)ELISA试剂盒是仅特异性测定Anit-OVA-IgG1的试剂盒。

 

◆特点

LBIS® 小鼠卵清蛋白特异性免疫球蛋白G1(OVA-IgG1)ELISA试剂盒                              LBIS® OVA-IgG1 Mouse


● 测定时间短(总反应时间:1小时50分钟)

● 微量样本即可测定。

● 使用无害的防腐剂。

● 全部试剂为溶液即用类型。

● 高测定精度和高重复性。

● 操作简便,无特殊前处理。

 



试剂盒组成


组成品

状态

包装

OVA包被96孔板(干燥板)

清洗后使用

96 wells(8×12)/1个

标准溶液(Anti OVA- IgG1:1,200U/mL)(单抗)

稀释后使用

100 μL/1瓶

缓冲液

直接使用

60 mL/1瓶

生物素结合抗小鼠IgG1抗体(单抗)

稀释后使用

200 μL/1瓶

过氧化物酶·抗生素结合物

稀释后使用

200 μL/1瓶

显色液(TMB)

直接使用

12 mL/1瓶

终止液(1M H2SO4

※小心轻放

直接使用

12 mL/1瓶

浓缩清洗液(10×)

稀释后使用

100 mL/1瓶

孔板密封膜

3个

产品说明书

1本

 


样本信息

● 小鼠血清·血浆

● 10 μL/well(稀释样本)

※ 样本必须用附带的缓冲液稀释100倍以上。

 


◆测定范围


1.88~120 mU/mL(标准曲线范围)

(本试剂盒中1 U/mL定义为抗原结合常数(Ka)为6.9×107 M-1的抗体160ng/mL)

 


实验数据


精密度实验(实验内变化)


样本

A

B

1

105

18.2

2

100

16.9

3

96.2

16.7

4

100

17.4

5

106

17.4

Mean

101

17.3

SD

4.0

0.59

CV(%)

4.0

3.4

单位:mU/mL



重复性实验(实验间变化)


测定日/样本

C

D

E

0天

60.1

15.0

3.75

1天

58.5

14.7

3.70

2天

59.9

15.3

3.83

3天

61.4

15.7

3.82

Mean

60.0

15.2

3.77

SD

1.2

0.44

0.06

CV(%)

2.0

2.9

1.6

单位:mU/mL



添加回收实验


样本F


添加量

实测值

回收量

回收率(%)

0.00

8.86

7.29

15.8

6.94

95.2

11.0

19.5

10.6

96.4

14.6

23.0

14.1

96.6

单位:mU/mL,n=3


样本G


添加量

实测值

回收量

回收率(%)

0.00

51.5

26.9

76.8

25.3

97.1

31.4

80.6

29.1

92.5

47.1

100

48.5

103

单位:mU/mL,n=3



稀释直线性实验


2个血清样本连续用稀释缓冲液稀释3个梯度测定结果,直线回归值R2=0.9994~0.9998

参考文献



 1.

Pinocembrin attenuates allergic airway inflammation via inhibition of NF-κB pathway in mice. Gu X, Zhang Q, Du Q, Shen H, Zhu Z. Int Immunopharmacol. 2017 Oct 18;53:90-95.


 2.

A dichloromethane fraction of Triticum aestivum sprouts reduces allergic immune response through inhibiting Th2 differentiation in ovalbumin‑immunized mice. Ki HH, Hwang SW, Lee JH, Kim YH, Kim DK, Lee YM. Mol Med Rep. 2017 Sep;16(3):3535-3541.


 3.

Urban PM2.5 exacerbates allergic inflammation in the murine lung via a TLR2/TLR4/MyD88-signaling pathway. He M, Ichinose T, Yoshida Y, Arashidani K, Yoshida S, Takano H, Sun G, Shibamoto T. Sci Rep. 2017 Sep 8;7(1):11027.


 4.

Activation of group 2 innate lymphoid cells exacerbates and confers corticosteroid resistance to mouse nasal type 2 inflammation. Morikawa T, Fukuoka A, Matsushita K, Yasuda K, Iwasaki N, Akasaki S, Fujieda S, Yoshimoto T. Int Immunol. 2017 May 1;29(5):221-233.


 5.

Aquaporin-3 potentiates allergic airway inflammation in ovalbumin-induced murine asthma. Ikezoe K, Oga T, Honda T, Hara-Chikuma M, Ma X, Tsuruyama T, Uno K, Fuchikami J, Tanizawa K, Handa T, Taguchi Y, Verkman AS, Narumiya S, Mishima M, Chin K. Sci Rep. 2016 May 11;6:25781.


 6.

Exposure to bisphenol A enhanced lung eosinophilia in adult male mice. He M, Ichinose T, Yoshida S, Takano H, Nishikawa M, Shibamoto T, Sun G. Allergy Asthma Clin Immunol. 2016 Apr 14;12:16.


 7.

Differences in allergic inflammatory responses between urban PM2.5 and fine particle derived from desert-dust in murine lungs. He M, Ichinose T, Kobayashi M, Arashidani K, Yoshida S, Nishikawa M, Takano H, Sun G, Shibamoto T. Toxicol Appl Pharmacol. 2016 Apr 15;297:41-55.


 8.

Desert dust induces TLR signaling to trigger Th2-dominant lung allergic inflammation via a MyD88-dependent signaling pathway. He M, Ichinose T, Song Y, Yoshida Y, Bekki K, Arashidani K, Yoshida S, Nishikawa M, Takano H, Shibamoto T, Sun G. Toxicol Appl Pharmacol. 2016 Apr 1;296:61-72.


 9.

Administration of Pigment Epithelium-Derived Factor Inhibits Airway Inflammation and Remodeling in Chronic OVA-Induced Mice via VEGF Suppression. Zha W, Su M, Huang M, Cai J, Du Q. Allergy Asthma Immunol Res. 2016 Mar;8(2):161-9.


10.

Low-dose benzo[a]pyrene aggravates allergic airway inflammation in mice. Yanagisawa R, Koike E, Win-Shwe TT, Ichinose T, Takano H. J Appl Toxicol. 2016 Feb 25.


11.

Prevention of allergic rhinitis by ginger and the molecular basis of immunosuppression by 6-gingerol through T cell inactivation. Kawamoto Y, Ueno Y, Nakahashi E, Obayashi M, Sugihara K, Qiao S, Iida M, Kumasaka MY, Yajima I, Goto Y, Ohgami N, Kato M, Takeda K. J Nutr Biochem. 2016 Jan;27:112-22.


12.

Immunotoxic Effect of Low-Dose Methylmercury Is Negligible in Mouse Models of Ovalbumin or Mite-Induced Th2 Allergy. Nakamura R, Takanezawa Y, Sone Y, Uraguchi S, Sakabe K, Kiyono M.

Biol Pharm Bull. 2016;39(8):1353-8.


13.

Prevention of allergic rhinitis by ginger and the molecular basis of immunosuppression by 6-gingerol through T cell inactivation. Yoshiyuki Kawamoto, Yuki Ueno, Emiko Nakahashi, Momoko Obayashi, Kento Sugihara, Shanlou Qiao, Machiko Iida, Mayuko Y. Kumasaka, Ichiro Yajima, Yuji Goto, Nobutaka Ohgami, Masashi Kato, Kozue Takeda. The Journal of Nutritional Biochemistry, Volume 27, Jan. 2016, Pages 112–122


14.

Effects of Sohamhyoong-Tang on Ovalbumin-Induced Allergic Reaction in BALB/c Mice. Jo SH, Lee YJ, Kang DG, Lee HS, Kim DK, Park MC. Evid Based Complement Alternat Med. 2016;2016:6286020.


15.

Effect of diosmetin on airway remodeling in a murine model of chronic asthma. Ge A, Liu Y, Zeng X, Kong H, Ma Y, Zhang J, Bai F, Huang M. Acta Biochim Biophys Sin (Shanghai). Vol.47(8), p604-11, Aug 2015.


16.

PM2.5-rich dust collected from the air in Fukuoka, Kyushu, Japan, can exacerbate murine lung eosinophilia. He M, Ichinose T, Ren Y, Song Y, Yoshida Y, Arashidani K, Yoshida S, Nishikawa M, Takano H, Sun G. Inhal Toxicol. Vol.27(6), p287-99, May 2015.


17.

Anti-asthma potential of crocin and its effect on MAPK signaling pathway in a murine model of allergic airway disease. Xiong Y, Wang J, Yu H, Zhang X, Miao C. Immunopharmacol Immunotoxicol. Vol.10, p1-8, Mar 2015.


18.

Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin. Kong X, Wu SH, Zhang L, Chen XQ. Mol Med Rep. 2015 Mar 5


19.

Pharyngeal aspiration of metal oxide nanoparticles showed potential of allergy aggravation effect to inhaled ovalbumin. Horie M, Stowe M, Tabei M, Kuroda E. Inhal Toxicol. Vol.27(3), p181-90, Feb 2015.


20.

Oxidized dietary oils enhance immediate- and/or delayed-type allergic reactions in BALB/c mice. Ogino H, Sakazaki F, Okuno T, Arakawa T, Ueno H. Allergol Int. Vol.64(1), p66-72, Jan 2015.


21.

The effects of nodakenin on airway inflammation, hyper-responsiveness and remodeling in a murine model of allergic asthma. Xiong Y, Wang J, Yu H, Zhang X, Miao C, Ma S. Immunopharmacol Immunotoxicol. Vol.36(5), p341-348, Oct 2014.


22.

Allogeneic pluripotent stem cells suppress airway inflammation in murine model of acute asthma. Ogulur I, Gurhan G, Kombak FE, Filinte D, Barlan I, Akkoc T. International Immunopharmacology, Vol.22(1), p31-40 Sep 2014.


23.

Effects of prior oral exposure to combinations of environmental immunosuppressive agents on ovalbumin allergen-induced allergic airway inflammation in Balb/c mice. Fukuyama T, Nishino R, Kosaka T, Watanabe Y, Kurosawa Y, Ueda H, Harada T.. Immunopharmacol Immunotoxicol. Vol.36(4), p261-70, Aug 2014.


24.

Enhancement of OVA-induced murine lung eosinophilia by co-exposure to contamination levels of LPS in Asian sand dust and heated dust. Ren Y, Ichinose T, He M, Song Y, Yoshida Y, Yoshida S, Nishikawa M, Takano H, Sun G, Shibamoto T. Allergy, Asthma & Clinical Immunology, Vol.10(1), Jun 2014.


25.

A bacterial extract of OM-85 Broncho-Vaxom prevents allergic rhinitis in mice. Han L, Zheng CP, Sun YQ, Xu G, Wen W, Fu QL. American Journal of Rhinology & Allergy, Vol.28(2), p110-116, Mar-Apr 2014.


26.

Broncho-Vaxom Attenuates Allergic Airway Inflammation by Restoring GSK3β-Related T Regulatory Cell Insufficiency. Fu R, Li J, Zhong H, Yu D, Zeng X, Deng M, Sun Y, Wen W, Li H. PLoS One. 2014 Mar 25;9(3):e92912


27.

Lung inflammation by fungus, Bjerkandera adusta isolated from Asian sand dust (ASD) aerosol and enhancement of ovalbumin-induced lung eosinophilia by ASD and the fungus in mice. Liu B, Ichinose T, He M, Kobayashi F, Maki T, Yoshida S, Yoshida Y, Arashidani K, Takano H, Nishikawa M, Sun G, Shibamoto T. Allergy, Asthma & Clinical Immunology, Vol.10(1), Feb 2014.


28.

Midazolam inhibits IgE production in mice via suppression of class switch recombination. Kusama H, Kobayashi R, Kurita-Ochiai T. Journal of Oral Science, Vol.56(1), p77-83, 2014.


29.

Induction of immune tolerance and reduction of aggravated lung eosinophilia by co-exposure to Asian sand dust and ovalbumin for 14 weeks in mice. He M., Ichinose T., Yoshida S., Takano H., Nishikawa M., Sun G. and Shibamoto T. Allergy, Asthma & Clinical Immunology, Vol.9(19), 2013.


30.

Galangin Abrogates Ovalbumin-Induced Airway Inflammation via Negative Regulation of NF-B. Zha W-J., Qian Y., Shen Y., Du Q., Chen F-F., Wu Z-Z., Li X. and Huang M. Evidence-Based Complementary and Alternative Medicine, Vol.2013 (2013), p14.


31.

Effects of two Asian sand dusts transported from the dust source regions of Inner Mongolia and northeast China on murine lung eosinophilia. M.He, T.Ichinose, Y.Song, Y.Yoshida, K.Arashidani, S.Yoshida, B.Liu, M.Nishikawa, H.Takano, G.Sun. Toxicology and Applied Pharmacology, Available online 26 July 2013.


32.

Effect of the size of receptor in allergy detection using field effect transistor biosensor. S.Hideshima, S.Kuroiwa, M.Kimura, S.Cheng, T.Osaka. Electrochimica Acta, Available online 24 July 2013.


33.

Elevated Macrophage Inflammatory Protein 1α and Interleukin-17 Production in an Experimental Asthma Model Infected with Respiratory Syncytial Virus. T.Ishioka, Y.Yamada, H.Kimura, M.Yoshizumi, H.Tsukagoshi, K.Kozawa, K.Maruyama, Y.Hayashi, M.Kato. Int Arch Allergy Immunol, Vol.161(suppl 2), p129-137, May 2013.


34.

Leukotriene B4 receptor BLT2 negatively regulates allergic airway eosinophilia . Y.Matsunaga, S.Fukuyama, T.Okuno, F.Sasaki, T. Matsunobu, Y.Asai, K.Matsumoto, K.Saeki, M.Oike, Y.Sadamura, K.Machida, Y.Nakanishi, M.Kubo, T.Yokomizo and H.Inoue. The FASEB Journal, Published online before print April 19, 2013.


35.

Effects of exposure to nanoparticle-rich or -depleted diesel exhaust on allergic pathophysiology in the murine lung. Tanaka M., Aoki Y., Takano H., Fujitani Y., Hirano S., Nakamura R., Sone Y., Kiyono M., Ichinose T., Itoh T., Inoue K. Journal of Toxicological Sciences, Vol.38(1), p35-48, Feb 2013.


36.

HIF-1α Inhibition Reduces Nasal Inflammation in a Murine Allergic Rhinitis Model. Zhou H, Chen X, Zhang W-M, Zhu L-P, Cheng L. PLOS one, 2012.


37.

Human Pluripotent Stem Cell-Derived Mesenchymal Stem Cells Prevent Allergic Airway Inflammation in Mice. Sun Y-Q, Deng M-X, He J, Zeng Q-X, Wen W, Wong D S.H, Tse H-F, Xu G, Lian Q, Shi J, Fu Q-L. STEM CELLS, Vol.30(12), p2692-2699, Dec 2012.


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WB DAB Chromogenic Kit (Mouse IgG)-Yellow WB DAB 显色试剂盒(小鼠 IgG)(黄色)


WB DAB Chromogenic Kit (Mouse IgG)-Yellow

WB DAB 显色试剂盒(小鼠 IgG)(黄色)

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WB DAB Chromogenic Kit (Mouse IgG)-Yellow                                                                                                                       WB DAB 显色试剂盒(小鼠 IgG)(黄色)


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