Ab7973

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab7973 is a recombinant monoclonal antibody specific for Human CXCR4. It is produced in HEK-293T cells.

Automatically generated - may contain errors

Lab products found in correlation

Ab32503, by Abcam (4 mentions) Ab62472, by Abcam (2 mentions) Ab2302, by Abcam (2 mentions) Cisplatin, by Merck Group (2 mentions) Sc 47778, by Santa Cruz Biotechnology (2 mentions) Ab8226, by Abcam (2 mentions) Nitrocellulose membrane, by Merck Group (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Df6087, by Affinity Biosciences (1 mentions) Af7014, by Affinity Biosciences (1 mentions) Ab9722, by Abcam (1 mentions) Ab2426, by Abcam (1 mentions) Ab48394, by Abcam (1 mentions) Biotinylated secondary antibody, by Vector Laboratories (1 mentions) Enhanced chemiluminescence, by Thermo Fisher Scientific (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Ab17185, by Abcam (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Ab16502, by Abcam (1 mentions) Bca protein assay kit, by CWBIO (1 mentions)

16 protocols using ab7973

Histopathological Analysis of Liver Injury

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Tissue samples were fixed, paraffin embedded, and sliced. Then the sections were deparaffinized, rehydrated, and stained with hematoxylin and eosin (H&E) to observe hepatic pathology. The H&E staining images were blindly analyzed to grade liver injury as previously reported (Tong et al., 2019 (link)). Briefly, liver injuries were assessed by following parameters, including cytoplasmic discoloration, vacuolation formation, nuclear pyknosis, nuclear fragmentation, nuclear discoloration, and red cell stasis. The liver injury severity is measured on a scale of 0-4 with higher score indicating more severe injury.
In addition, paraffinized liver sections were also studied with immunohistochemistry (IHC) staining. After blocking with 10% goat serum, the sections were incubated with the primary antibodies against TNF-α (AF7014, 1:200, Affinity), IL-6 (DF6087, 1:200, Affinity), IL-1β (ab9722, 1:500, Abcam), Bcl-2 (ab7973, 1:100, Abcam), or Bax (50599-2-lg, 1:500, Proteintech) overnight at 4 °C. Following that, the secondary antibody incubation and DAB color rendering were performed, and the images were visualized under a microscope. In addition, quantitative analysis was conducted using Image J software.

Lu R., Yu R.J., Yang C., Wang Q., Xuan Y., Wang Z., He Z., Xu Y., Kou L., Zhao Y.Z., Yao Q, & Xu S.H. (2022). Evaluation of the hepatoprotective effect of naringenin loaded nanoparticles against acetaminophen overdose toxicity. Drug Delivery, 29(1), 3256-3269.

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Immunohistochemical Analysis of Cell Signaling

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Mounted paraffin sections were dewaxed in xylene, rehydrated in decreasing graded alcohols and washed in tap water. Endogenous peroxidase activity was inhibited in tissue sections using 0.5% v/v H2O2/methanol for 20 min at room temperature. Then, sections were blocked for 1 h with 15% normal goat serum in phosphate buffered saline (PBS) and then incubated overnight at 4°C with the primary antibody rabbit anti-PCNA (1:200 diluted ab2426, Abcam, UK); rabbit anti-BAX (1:200 diluted P-19, sc-526, Santa Cruz Biotechnologies, USA); rabbit anti-BCL2 (1:100 diluted ab7973, Abcam, UK); rabbit anti-CASPASE 3 (1:300 diluted AF835, RyD Systems, USA); rabbit anti-BECLIN 1 (1:500 diluted ab62472, Abcam, UK) or rabbit anti-LC3B (1:200 diluted ab48394, Abcam, UK). The, sections were rinsed in PBS and incubated for 1 h at room temperature with the appropriate 1:200 diluted biotinylated secondary antibody (Vector Labs, Peterborough, UK). After further washing in PBS, sections were incubated for 30 min with 1:100 diluted streptavidin-peroxidase complex (ABC kit, Vector Labs, Peterborough, UK). Finally, development of peroxidase activity was performed with 0.05% w/v 3,3’-diaminobenzidine and 0.1% v/v H2O2 in Tris-HCl. Negative controls were processed simultaneously by omitting the primary antibodies or pre-absorbing the primary antibody with specific synthetic peptides.

González C.R., Muscarsel Isla M.L, & Vitullo A.D. (2018). The balance between apoptosis and autophagy regulates testis regression and recrudescence in the seasonal-breeding South American plains vizcacha, Lagostomus maximus. PLoS ONE, 13(1), e0191126.

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Western Blot Analysis of Apoptosis Regulators

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The cells were lysed in RIPA lysis buffer (Beyotime). The lysates were centrifuged at 12,000 rpm and 4 °C for 10 min. The same amounts of protein were separated using 10–15% odium dodecyl sulfate-polyacrylamide gel electrophoresis (the exact concentration was determined by the molecular weight of the detected proteins) and transferred to nitrocellulose membranes (Millipore). For immunodetection, the membranes were incubated with specific antibodies and the following antibodies were used: anti-Bcl2 (Abcam, ab7973), anti-BAX (Abcam, ab7977), anti-caspase 3(CST, 9662S), anti-HO-1(CST, 5853S), anti-ERK (Abcam, ab17942), anti-p-ERK(CST, 5683S), anti-JNK(CST, 9258S), anti-p-JNK(CST, 9255S), anti-AKT(CST, 4691S), anti-p-AKT(CST, 4060S), anti-P38(CST, 9212S), anti-p-P38(CST, 4511S). The immunoblots were developed using horseradish peroxidase (HRP)-coupled anti-rabbit secondary antibodies (ProteinTech Group) and then detected with enhanced chemiluminescence (Pierce Biotechnology). The GAPDH protein was used as a control.

Li D., Tong W., Liu D., Zou Y., Zhang C, & Xu W. (2017). Astaxanthin mitigates cobalt cytotoxicity in the MG-63 cells by modulating the oxidative stress. BMC Pharmacology & Toxicology, 18, 58.

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Western Blot Analysis of Heart Proteins

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Heart samples were drawn at 2 h after reperfusion and detected as previously described [17 (link)]. Frozen heart tissue was homogenized in lysis buffer. The protein concentration was determined using a BCA Protein assay kit (Cwbiotech, China). Next, 2 µg/µl of protein was separated using 10% SDS-PAGE and then electrophoretically transferred onto PVDF membranes (Millipore, USA). Proteins on the membranes were then probed using primary antibodies, including mouse Bcl₂ (ab7973, abcam), Bax (ab32503, abcam), Caspase-3 (ab17185, abcam), NF-κBp65 (ab16502, abcam), and GAPDH (5174, CST). Following incubation with secondary antibodies, including goat anti-rabbit secondary antibody, the positive protein blots were developed using a chemiluminescent system, and the bands were quantified using the Gel Image system ver. 4.00 (Tanon, China). Data were shown as the percentage density of the blots.

Yuan R., Guo L.L., Chen H.W., Li J.P., Chen Z., Wei B.J, & Wang J. (2017). Cardioprotective Effects of HuoxueAnshen Recipe against Myocardial Injuries Induced by Sleep Deprivation in Rats. Evidence-based Complementary and Alternative Medicine : eCAM, 2017, 7359760.

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Hypoxia-Induced Cardiomyocyte Protein Profiling

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Eight hours after hypoxia, protein was isolated from cardiomyocytes with standard Invitrogen protocols (Invitrogen, Carlsbad, CA, USA). Protein concentration quantitation was modified by Bradford assay (Bio‐Rad Laboratories, Hercules, CA, USA), protein was then separated by SDSPAGE with antibodies against Mst1 (ab51134; Abcam), p‐Mst1 (Thr183) (#110687; Sigma‐Aldrich), LC3A/B (#12741S, CST), P62 (ab91526; Abcam), Beclin1 (ab62472; Abcam), GAPDH (ab181602; Abcam), Caspase‐3(ab4051; Abcam), Cleaved caspase‐3 (ab2302; Abcam), Bax (ab5714; Abcam), Bcl‐2 (ab7973; Abcam). The blots were visualized with a chemiluminescene system (Amersham Bioscience, Buchinghamshire, UK). The signals were quantified by Image Pro Plus software (Media Cybernetics, MD Rockville, USA).

Hu J., Man W., Shen M., Zhang M., Lin J., Wang T., Duan Y., Li C., Zhang R., Gao E., Wang H, & Sun D. (2015). Luteolin alleviates post‐infarction cardiac dysfunction by up‐regulating autophagy through Mst1 inhibition. Journal of Cellular and Molecular Medicine, 20(1), 147-156.

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Antibody Panel for Cell Signaling

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The monoclonal antibodies (mAbs) and polyclonal antibodies (pAbs) used in this study were as follows: Anti-MIF (mAb, ab55445), Anti-ERK1/2 (phospho T185+Y187+T202+T204, pAb, ab50011), anti-Bax (pAb, ab7977), anti-HMGB1 (pAb, ab79823), anti-Bcl-2 (pAb, ab7973) and anti-Histone H3 (pAb, ab1791) were purchased from Abcam (Cambridge, MA, USA). Anti-beta-actin (pAb) and anti-GADPH (pAb) was purchased from Anbo Biotechnology Company (San Francisco, CA, USA). Anti-PARP and anti-LC3B (pAb) was purchased from Cell Signaling Technology (Beverly, MA, USA). AnnexinV-FITC and propidium iodide were purchased from BD Pharmingen (San Jose, CA, USA).
Doxorubicin, cisplatin, 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), 2′,7′-dichlorofluorescin diacetate (DCF-DA) and 3-Methyladenine (3-MA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

Liu Y., Zhao L., Ju Y., Li W., Zhang M., Jiao Y., Zhang J., Wang S., Wang Y., Zhao M., Zhang B, & Zhao Y. (2014). A novel androstenedione derivative induces ROS-mediated autophagy and attenuates drug resistance in osteosarcoma by inhibiting macrophage migration inhibitory factor (MIF). Cell Death & Disease, 5(8), e1361-.

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Thrombolysis Effect on Iliac Artery

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To confirm the existence of thrombi in vivo and the effect of thrombolysis, iliac artery specimens from the rats were embedded in paraffin and sliced (5 µm). The sliced samples were deparaffinized and stained with hematoxylin and eosin (H&E). Then, immunohistochemical staining was performed on deparaffinized tissue sections with water bath–induced antigen retrieval. Endogenous peroxidase was inactivated by incubating in 3% H₂O₂ (containing 80% methanol) for 10 min. Nonspecific binding was blocked by 5% goat serum for 45 min. The primary antibodies, including anti-Caspase-3 antibody (a11593, ABclonal), anti-Caspase-12 antibody (ab62484, Abcam), anti-BAX-antibody (50599-2-Ig, Proteintech), and anti-Bcl-2 antibody (ab7973, Abcam), were added to the sections overnight at 4°C. The sections were washed with PBS and covered with horseradish peroxidase (HRP)–conjugated secondary goat anti-rabbit IgG for 1 h at 37°C. Finally, the cells were stained with diaminobenzidine (DAB) and counterstained with hematoxylin.

Liang Z., Chen H., Gong X., Shi B., Lin L., Tao F., Wu Q., Fang M., Li H., Lu C., Xu H., Zhao Y, & Chen B. (2022). Ultrasound-Induced Destruction of Nitric Oxide–Loaded Microbubbles in the Treatment of Thrombus and Ischemia–Reperfusion Injury. Frontiers in Pharmacology, 12, 745693.

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Western Blot Analysis of Protein Lysates

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Protein lysates from tissue samples were obtained using RIPA buffer supplemented with protease and phosphatase inhibitors. For the cellular samples, cells were collected following treatment with DMSO and compounds. Western blot was performed as previously described.^{25 (link)} The primary antibodies were BRD2 (A302-583A, Bethyl), BRD3 (A302-368A, Bethyl), BRD4 (A301-985A, Bethyl), MYC (sc-40, Santa Cruz Biotechnology), BCL2 (ab7973, Abcam), BAX (ab32503, Abcam), cleaved PARP (Cell Signaling, #5625). The secondary antibodies were anti-rabbit IgG (Jackson, 111-035-003) or anti-mouse IgG (Jackson). HRP substrate (GE Healthcare) was used for the detection of chemical luminescence, and FUJIFILM LAS-4000 was used for image acquisition.

Chen W., Zhang H., Chen Z., Jiang H., Liao L., Fan S., Xing J., Xie Y., Chen S., Ding H., Chen K., Jiang H., Luo C., Zheng M., Yao Z., Huang Y, & Zhang Y. (2018). Development and evaluation of a novel series of Nitroxoline-derived BET inhibitors with antitumor activity in renal cell carcinoma. Oncogenesis, 7(11), 83.

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Protein Expression Analysis via Western Blot

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Total protein was extracted according to the instructions of the kit, and protein concentration was measured by the BCA protein quantification method (no. 23227; Pierce, Rockford, IL, USA). Protein samples were stored at −70°C before use. Electrophoresis was performed using Nu-PAGE 10% SDS-PAGE Bis-Tris gel in SDS-PAGE buffer. Polyvinylidene fluoride (PVDF) membrane was used for transfer. Next, the membrane was blocked with bovine serum albumin (3%). Afterwards, the membranes were washed with Tris-buffered saline containing tween 20 (TBST) and incubated overnight with primary antibodies (procaspase-3 (ab184787; Abcam Inc., Cambridge, MA, USA), procaspase-9 (ab184786; Abcam Inc., Cambridge, MA, USA), anti-Bcl-2 antibody (ab7973; Abcam Inc., Cambridge, MA, USA), anti-Bax (ab32503; Abcam Inc., Cambridge, MA, USA),and β-actin (sc-47778; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA)) diluted 1 : 1000. After that, the membrane was washed three times with TBST and the secondary antibody (1 : 1000) was added to be incubated for 1 h and washed with TBST. Then, band intensities were detected using the chemiluminescent substrate Supersignal Femto kit and band densities were analyzed using the ImageJ 1.52a program (Bethesda, Maryland, USA) [27 (link)].

Azimi M., Mehrzad J., Ahmadi E., Orafei M., Aghaie F., Ahmadi A., Rahimi M, & Ghorbani Ranjbary A. (2022). The Effect of Thymus vulgaris on Hepatic Enzymes Activity and Apoptosis-Related Gene Expression in Streptozotocin-Induced Diabetic Rats. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 2948966.

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Western Blot Analysis of Inflammatory Markers

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Western blot was performed according to our previously protocols [28 (link)]. Briefly, the brain tissue was homogenized and centrifuged. The detergent compatible protein assay kit (Bio-Rad, Hercules, CA, USA) was used to determine the protein content. Equal amounts of protein (60 μg) were loaded into the wells of the SDS-PAGE gel. After electrophoresis, the protein was transferred to a nitrocellulose membrane. The membrane was blocked for 2 h using a nonfat dry milk buffer and incubated overnight at 4 °C with the primary antibodies. The following primary antibodies were used: rabbit polyclonal anti-beclin 1 (1:800, ab62557, Abcam), rabbit monoclonal anti-NLRP3(1:1000, ab210491, Abcam), goat polyclonal anti-caspase-1 (1:2000, sc-22165, Santa Cruz), rabbit polyclonal anti-IL-1ß (1:200, ab7973, Abcam), rabbit polyclonal anti-IL-18 (1:1000, cst# 9272, Cell Signaling Technology), and β-actin (1:4000, sc-47778Santa Cruz). The membrane was incubated for 1 h with secondary antibodies (horseradish peroxidase-conjugated) at room temperature. The membrane was exposed to X-ray film, and band densities were analyzed using Image J software. To facilitate comparisons between groups, band density values were normalized to the mean value for the sham group.

Li J.R., Xu H.Z., Nie S., Peng Y.C., Fan L.F., Wang Z.J., Wu C., Yan F., Chen J.Y., Gu C., Wang C., Chen J.S., Wang L, & Chen G. (2017). Fluoxetine-enhanced autophagy ameliorates early brain injury via inhibition of NLRP3 inflammasome activation following subrachnoid hemorrhage in rats. Journal of Neuroinflammation, 14, 186.

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