The STORM super-resolution (SR) imaging of NIH/3T3 cells using [KD Validated] LAMP1/CD107a Rabbit PolymAb®（A24804PM, Abclonal at dilution of 1:200 with 3% paraformaldehyde (PFA) +0.1% glutaraldehyde (GA) fixation. The immunostaining was performed by Full Automatic Immunofluorescence Workflow System (Workflow Ultra300, Nano-Micro imaging, China). Image was performed with Single-Molecule Localization Super-Resolution Microscopy (STORM Ultra300, Nano-Micro imaging, China). We acknowledge Nano-Micro imaging Biotechnology Co., Ltd. (宁波纳微成像生物科技有限公司) in SR image processing and kindly providing this image.
Western blot analysis of lysates from NIH/3T3 cells using [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM) at 1:5000 dilution incubated at room temperature for 1.5 hours. NIH/3T3 cells were treated with PNGase F(1μL) at 37℃ for 3 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 Mouse kidney using [KD Validated] LAMP1/CD107a Rabbit PolymAb® at 1:5000 dilution incubated at room temperature for 1.5 hours.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: 10s.
Western blot analysis of lysates from U-251 MG cells using [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM) at 1:5000 dilution incubated at room temperature for 1.5 hours.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: 45s.
Western blot analysis of lysates from wild type (WT) and LAMP1/CD107a knockdown (KD) HeLa cells using [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM) at 1:5000 dilution incubated at room temperature for 1.5 hours.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: 90s.
Confocal imaging of NIH/3T3 cells using [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM, 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 and LAMP1/CD107a knockdown(KD) HeLa cells using [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM, 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 C2C12 cells using [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM, 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.
Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM) at a dilution of 1:500 (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 [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM) at a dilution of 1:500 (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 [KD Validated] LAMP1/CD107a Rabbit PolymAb® (A24804PM) at a dilution of 1:500 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer (pH 9.0) prior to IHC staining.
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Enables protein domain specific binding activity. Involved in protein stabilization. Located in several cellular components, including cytoplasmic vesicle; sarcolemma; and vacuole. Is expressed in several structures, including 1-cell stage embryo; central nervous system; craniocervical region bone; extraembryonic component; and gut. Orthologous to human LAMP1 (lysosomal associated membrane protein 1).
首先，一般抗体不推荐客户回收利用，抗体使用之后缓冲体系已经发生改变，不同客户在回收抗体的保存条件上也会有差异，所以抗体回收使用效果无法保证。另外，ABclonal公司也做过一批抗体回收验证测试，测试结果显示不同抗体可回收次数不同，一般效价越高的抗体，可重复使用的次数越多，客户可根据实验情况来确定。
注：我们将孵育完毕后剩余的抗体回收到离心管中置于4℃保存，效价高的抗体可至少保存1周，至少重复利用3次。
武汉爱博泰克生物（ABclonal）科技有限公司是国产品牌，她成立于2011年，公司依托ABclonal美国波士顿抗体与蛋白研发中心、中国光谷生物城(武汉)抗体生产基地以及上海张江分子酶研发中心，凝聚了十余位来自哈佛大学、麻省理工、复旦大学、上海交大、中科院生化细胞所和武汉大学的全球知名分子和免疫学方面博士，组成我们的科学家团队，通过聚焦抗体与酶核心技术，致力于打破国际技术的垄断，将公司打造成为科研工具和诊断原料的国内领导品牌，乃至弯道超越国际巨头。 我们拥有包括兔多克隆抗体、小鼠单克隆抗体、兔单克隆抗体的生产研发平台，同时也有包括WB,IHC,IF,IP,CHIP在内的检测平台，我们对每一支自产的抗体进行了严格的检测。当然，我们部分直销地区也可以帮客户代购进口品牌的产品。同时也有抗体定制服务。ABclonal抗体优势：1，严自检，保质量；2产品多，指标全；3，价格低，货期短。注：ABclonal抗体价格体系详情见附件
ABclonal抗体成分在平时工作当中，常会有客户咨询我们的抗体用的什么buffer进行保存，一般来说，我们的buffer的成分是：PBS含0.03%的proclin300、0.05%牛血清白蛋白、50%甘油；也有一些是PBS含0.03%的proclin300，50%甘油。防腐剂 Proclin 300活性成分主要是2-甲基-4-异噻唑啉-3-酮（MCI）和5-氯-2-甲基-4-异噻唑啉-3-酮（CMCI）。ProClin生物灭活剂能够迅速穿透细胞膜，抑制对细胞呼吸至关重要的特定酶，因此一接触微生物有机体就会立即抑制细胞活性。ProClin的多个特定毒性位点可以防止微生物产生高水平的耐药性。
