BCL2 Monoclonal antibody

• Featured Product
• KD/KO Validated

BCL2 Monoclonal Antibody for FC, IF, IHC, IP, WB, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, pig and More (1)

Applications

WB, IP, IHC, IF, ICC, FC, CoIP, ELISA

Conjugate

Unconjugated

Publications

214

CloneNo.

4H8C6

Cat no : 60178-1-Ig

Synonyms

Apoptosis regulator Bcl 2, B cell CLL/lymphoma 2, BCL2, BCL-2, Oncogene B cell leukemia 2

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Validation Data Gallery

• 

  WB analysis using 60178-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Jurkat cells using 60178-1-Ig

  WB result of BCL2 antibody (60178-1-Ig, 1:1000) with si-control and si-BCL2 transfected Jurkat cells.

• 

  WB analysis of U2OS using 60178-1-Ig

  U2OS cells were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 60178-1-Ig

  MCF-7 cells were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 60178-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:15000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 60178-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 60178-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of pig spleen using 60178-1-Ig

  pig spleen tissue were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 60178-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of THP-1 using 60178-1-Ig

  THP-1 cells were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of U-937 using 60178-1-Ig

  U-937 cells were subjected to SDS PAGE followed by western blot with 60178-1-Ig (BCL2 Antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of U-937 using 60178-1-Ig

  IP Result of anti-BCL2 (IP:60178-1-Ig, 5ug; Detection:60178-1-Ig 1:1000) with U-937 cells lysate 4000ug.

• 

  IHC staining of human appendicitis using 60178-1-Ig

  Immunohistochemical analysis of paraffin-embedded human appendicitis tissue slide using 60178-1-Ig (BCL2 antibody) at dilution of 1:12800 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human appendicitis using 60178-1-Ig

  Immunohistochemical analysis of paraffin-embedded human appendicitis tissue slide using 60178-1-Ig (BCL2 antibody) at dilution of 1:12800 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of Jurkat using 60178-1-Ig

  Immunohistochemical analysis of paraffin-embedded Jurkat cells slide using 60178-1-Ig (BCL2 antibody) at dilution of 1:10000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lymphoma using 60178-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lymphoma tissue slide using 60178-1-Ig (BCL2 antibody) at dilution of 1:12800 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lymphoma using 60178-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lymphoma tissue slide using 60178-1-Ig (BCL2 antibody) at dilution of 1:12800 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 60178-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 60178-1-Ig (BCL2 Antibody) at dilution of 1:1000 (under 10x lens).

• 

  IHC staining of human tonsillitis using 60178-1-Ig

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 60178-1-Ig (BCL2 Antibody) at dilution of 1:1000 (under 40x lens).

• 

  IF Staining of MCF-7 using 60178-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed MCF-7 cells using BCL2 antibody (60178-1-Ig, Clone: 4H8C6 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of human appendicitis using 60178-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human appendicitis tissue using BCL2 antibody (60178-1-Ig, Clone: 4H8C6 ) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of human tonsillitis using 60178-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using BCL2 antibody (60178-1-Ig, Clone: 4H8C6 ) at dilution of 1:1500 and CoraLite®594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of human peripheral blood lymphocytes using 60178-1-Ig

  1X10^6 human peripheral blood lymphocytes were stained with 0.2 ug Anti-Human BCL2 (60178-1-Ig, Clone:4H8C6) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (green), or 0.2 ug isotype control and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (black). Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100.

• 

  FC experiment of Jurkat using 60178-1-Ig

  1X10^6 Jurkat cells were stained with 0.2ug BCL2 antibody (60178-1-Ig, red) and control antibody (blue). Fixed with 90% MeOH blocked with 3% BSA (30 min). Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L) with dilution 1:1000.

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Tested Applications

• Positive WB detected in: U2OS cells, HEK-293 cells, HepG2 cells, Jurkat cells, MCF-7 cells, pig spleen tissue, THP-1 cells, U-937 cells, MOLT-4 cells, HL-60 cells, HT-1080 cells, Raji cells
• Positive IP detected in: U-937 cells
• Positive IHC detected in: human appendicitis tissue, human tonsillitis tissue, human lymphoma tissue, Jurkat cells; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: MCF-7 cells, human tonsillitis tissue, human appendicitis tissue
• Positive FC detected in: human peripheral blood lymphocytes, Jurkat cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for IP and 1:500-1:2000 for WB
• Immunohistochemistry (IHC): IHC : 1:6400-1:25600
• Immunofluorescence (IF): IF : 1:200-1:800

Sample-dependent, check data in validation data gallery

Published Applications

• WB: See 200 publications below
• IHC: See 13 publications below
• IF: See 7 publications below
• CoIP: See 2 publications below

Product Information

60178-1-Ig targets BCL2 in WB, IP, IHC, IF, ICC, FC, CoIP, ELISA applications and shows reactivity with human, pig samples.

• Tested Reactivity: human, pig
• Cited Reactivity: human, monkey, pig
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: BCL2 fusion protein Ag3508
• Full Name: B-cell CLL/lymphoma 2
• Calculated molecular weight: 26 kDa
• Observed molecular weight: 26 kDa
• GenBank accession number: BC027258
• Gene symbol: BCL2
• Gene ID (NCBI): 596
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

1) What is Bcl-2?

Bcl-2 (B-cell lymphoma 2) is an outer mitochondrial membrane protein that via heterodimerization with BAX proteins controls apoptosis. Abnormalities of Bcl-2 activity can lead to cancer, autoimmune diseases, and schizophrenia.

2) FAQs for Bcl-2

A) How to measure Bcl-2 and Bax apoptotic markers by western blotting
Generally, Bcl-2 protein is considered to have anti-apoptotic activity, while Bax has apoptotic activity. Comparing the ratio between Bcl-2 and Bax protein levels in different samples or cells subjected to treatments can reflect the apoptotic state of the cells. Expression of Bax can vary between cell lines and it is important to compare it to Bcl-2 levels.
B) What band size should I expect in western blotting for Bcl-2?
The molecular weight of Bcl-2 is 26 kDa.

Publications

Species	Application	Title
human	WB	J Extracell Vesicles; Extracellular vesicles derived from oesophageal cancer containing P4HB promote muscle wasting via regulating PHGDH/Bcl-2/caspase-3 pathway.; Authors - Xiaohan Gao
rat	WB	Redox Biol; Heat shock protein 22 modulates NRF1/TFAM-dependent mitochondrial biogenesis and DRP1-sparked mitochondrial apoptosis through AMPK-PGC1α signaling pathway to alleviate the early brain injury of subarachnoid hemorrhage in rats.; Authors - Haiyan Fan
human	WB	Redox Biol; Mecheliolide elicits ROS-mediated ERS driven immunogenic cell death in hepatocellular carcinoma.; Authors - Zhongren Xu
human	WB	J Immunother Cancer; Switch receptor T3/28 improves long-term persistence and antitumor efficacy of CAR-T cells; Authors - Songbo Zhao
monkey	CoIP	Autophagy; Bunyavirus SFTSV exploits autophagic flux for viral assembly and egress; Authors - Jia-min Yan
human	WB	Autophagy; TSPAN1 promotes autophagy flux and mediates cooperation between WNT-CTNNB1 signaling and autophagy via the MIR454-FAM83A-TSPAN1 axis in pancreatic cancer.; Authors - Cefan Zhou

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